Publications by category
Books
Zhang J, Labes A, Zeng R (2021). Book: Marine Microbial-Derived Molecules and Their Potential Medical and Cosmetic Applications., Lausanne: Frontiers Media SA.
(2021). Neuroprotection: Rescue from Neuronal Death in the Brain., MDPI.
Journal articles
Pracucci E, Graham R, S Alberio L, Nardi G, Cozzolino O, Pillai V, Saieva L, Walsh D, Landi S, Zhang J, et al (In Press). Circadian rhythm in cortical chloride homeostasis underpins variation in network excitability. bioRxiv
Chen Z, Zhang J, Heilig R, Sorrell FJ, D’Angiolella V, Fischer R, Alessi DR, Bullock AN (In Press). Sequence and structural variations determining the recruitment of WNK kinases to the KLHL3 E3 ligase.
Abstract:
Sequence and structural variations determining the recruitment of WNK kinases to the KLHL3 E3 ligase
AbstractThe BTB-Kelch protein KLHL3 is a Cullin3-dependent E3 ligase that mediates the ubiquitin-dependent degradation of kinases WNK1-4 to control blood pressure and cell volume. A crystal structure of KLHL3 has defined its binding to an acidic degron motif containing a PXXP sequence that is strictly conserved in WNK1, WNK2 and WNK4. Mutations in the second proline abrograte the interaction causing the hypertension syndrome pseudohypoaldosteronism type II. WNK3 shows a diverged degron motif containing 4 amino acid substitutions that remove the PXXP motif raising questions as to the mechanism of its binding. To understand this atypical interaction, we determined the crystal structure of the KLHL3 Kelch domain in complex with a WNK3 peptide. The electron density enabled the complete 11-mer WNK-family degron motif to be traced for the first time revealing several conserved features not captured in previous work, including additional salt bridge and hydrogen bond interactions. Overall, the WNK3 peptide adopted a conserved binding pose except for a subtle shift to accommodate bulkier amino acid substitutions at the binding interface. At the centre, the second proline was substituted by WNK3 Thr541, providing a unique phosphorylatable residue among the WNK-family degrons. Fluorescence polarisation and structural modelling experiments revealed that its phosphorylation would abrogate the KLHL3 interaction similarly to hypertension-causing mutations. Together, these data reveal how the KLHL3 Kelch domain can accommodate the binding of multiple WNK isoforms and highlight a potential regulatory mechanism for the recruitment of WNK3.
Abstract.
Delmotte Q, Medina I, Hamze M, Buhler E, Zhang J, Belgacem YH, Porcher C (In Press). Smoothened receptor Signaling regulates the developmental shift of GABA polarity in rat somatosensory cortex.
Abstract:
Smoothened receptor Signaling regulates the developmental shift of GABA polarity in rat somatosensory cortex
ABSTRACTSonic Hedgehog (Shh) and its patched-smoothened receptor complex control a variety of functions in the developing central nervous system (CNS) such as neural cell proliferation and differentiation. Recently, Shh signaling components have been found to be expressed at the synaptic level in the postnatal brain, suggesting a potential role in the regulation of synaptic transmission. Usingin uteroelectroporation of constitutively active and dominant-negative forms of the Shh co-receptor smoothened (Smo), we studied the role of Smo signaling in the development and maturation of GABAergic transmission in the somatosensory cortex. Our results show that enhancing Smo activity during development accelerates the shift from depolarizing to hyperpolarizing GABA in dependence on functional expression of potassium-chloride cotransporter type 2 (KCC2). On the other hand, blocking Smo activity maintains GABA response in a depolarizing state in mature cortical neurons resulting in altered chloride homeostasis and increased seizure susceptibility. This study reveals an unexpected function of Smo signaling on the regulation of chloride homeostasis through the control of KCC2 cell surface stability and on the timing of the GABA inhibitory/excitatory shift in brain maturation.Summary statementThe smoothened receptor controls the time course of inhibitory transmission through the stability of the potassium-chloride cotransporter type 2 at the plasma membrane.
Abstract.
Zhang J, Wang J, Labes A, Zeng R (2023). Editorial: Marine microbial-derived molecules and their potential medical and cosmetic applications, volume II. Frontiers in Microbiology, 14
Zhang J, Siew K, Sun D (2023). Editorial: Targeting pumps, channels and transporters for the treatments of vascular, cardiovascular and kidney diseases. Frontiers in Pharmacology, 14
Bhuiyan MIH, Fischer S, Patel SM, Oft H, Zhang T, Foley LM, Zhang J, Hitchens TK, Molyneaux BJ, Deng X, et al (2023). Efficacy of novel SPAK inhibitor ZT-1a derivatives (1c, 1d, 1g & 1h) on improving post-stroke neurological outcome and brain lesion in mice. Neurochemistry International, 162, 105441-105441.
Zhang J (2023). Oral Paclovid Significantly Reduces Hospitalization and Mortality in Non-hospitalized Elderly Patients with COVID-19.
Journal of Modern Biology and Drug DiscoveryAbstract:
Oral Paclovid Significantly Reduces Hospitalization and Mortality in Non-hospitalized Elderly Patients with COVID-19
The development of antiviral drugs against coronavirus has proceeded at an unprecedented pace. In December 2021, the US FDA has successively approved two oral drugs against coronavirus pneumonia (COVID-19), namely PAXLOVID™ (PF-07321332) developed by Pfizer and Molnupiravir developed by Merck by targeting papain-like protease (PLpro) and RNA-dependent RNA polymerase (RdRp), respectively. PLpro and RdRp are important for the normal life cycle of coronaviruses. Inhibition of their activities could impair the synthesis of viral RNA and aid therapeutic treatments. However, little is known about the real-world effectiveness of oral antivirals against the severe acute respiratory syndrome (SARS)-CoV-2 omicron (B.1.1.529) variant. Recent studies have shown that Paxlovid can lead to a marked reduction hospitalization or death among unvaccinated outpatients with early COVID-19, whereas it was found no significant benefit for patients aged 64 and younger. In comparison, Molnupiravir did not reduce the risk of hospitalization or death after infection in high-risk groups who had been vaccinated against the coronaviruses, but only accelerated their recovery.
Abstract.
Robert SM, Reeves BC, Kiziltug E, Duy PQ, Karimy JK, Mansuri MS, Marlier A, Allington G, Greenberg ABW, DeSpenza T, et al (2023). The choroid plexus links innate immunity to CSF dysregulation in hydrocephalus. Cell, 186(4), 764-785.e21.
Robert S, Reeves B, Karimy JK, Marlier A, Kiziltug E, DeSpenza T, Singh A, Allington G, Phan D, Zhang J, et al (2022). 374 Multi-omic Analysis Identifies a SPAK Kinase-regulated Ensemble of Choroid Plexus Ion Transport Proteins Relevant for Post-infectious Hydrocephalus. Neurosurgery, 68(Supplement_1), 89-89.
D’Silva E, Meor Azlan NF, Zhang J (2022). Angiotensin II Receptor Blockers in the Management of Hypertension in Preventing Cognitive Impairment and Dementia—A Systematic Review.
Pharmaceutics,
14(10), 2123-2123.
Abstract:
Angiotensin II Receptor Blockers in the Management of Hypertension in Preventing Cognitive Impairment and Dementia—A Systematic Review
Hypertension is a known risk factor for cognition-related pathologies including dementia. The National Institute of Health and Care Excellence (NICE) guidelines recommend angiotensin (Ang) II receptor blockers (ARBs) or angiotensin-converting enzyme inhibitors (ACEIs) as a first-line treatment for hypertension. Although both ARBs and ACEIs show neuroprotective effects, ACEIs show contradictory side effects; therefore, ARBs may be a more viable option. However, trials assessing the effects of ARBs on cognition are scarce and conflicting. Therefore, the aim of this review is to conduct a systematic review and synthesise data on the influence of ARBs on cognition and dementia prevention. Five databases were searched from 1992–2022 to produce 13 randomised controlled trials (RCTs) involving 26,907 patients that compared associations of ARBs against placebos or other antihypertensives on cognition or probable dementia with a minimum duration of 3 months. ARBs showed greater cognitive benefits when compared to hydrochlorothiazide (HCTZ), beta blockers (BB), and ACEIs. Our findings showed that although ARBs are superior to some antihypertensives such as ACEIs, thiazide and beta blockers, they made no difference in comparison to the placebo in all but one sample of patients. The positive effects on cognitive performances are equal to calcium channel blockers (CCBs) and lower than statin. The neuroprotective effects of ARBs are also more beneficial when ARBs are taken at the same time as a statin. Due to these inconsistencies, robust conclusions cannot be made. Future trials are warranted and, if successful, could have positive economic implications and consequently improve quality of life.
Abstract.
Zhang J, Yao J, Rong M (2022). Editorial: Role of Ion Channels in Pain. Frontiers in Pharmacology, 13
Chen L, Yu J, Wan L, Wu Z, Wang G, Hu Z, Ren L, Zhou J, Qian B, Zhao X, et al (2022). Furosemide prevents membrane KCC2 downregulation during convulsant stimulation in the hippocampus. IBRO Neuroscience Reports, 12, 355-365.
Bhuiyan MIH, Young CB, Jahan I, Hasan MN, Fischer S, Meor Azlan NF, Liu M, Chattopadhyay A, Huang H, Kahle KT, et al (2022). NF-κB Signaling-Mediated Activation of WNK-SPAK-NKCC1 Cascade in Worsened Stroke Outcomes of Ang II–Hypertensive Mice.
Stroke,
53(5), 1720-1734.
Abstract:
NF-κB Signaling-Mediated Activation of WNK-SPAK-NKCC1 Cascade in Worsened Stroke Outcomes of Ang II–Hypertensive Mice
. Background:
. Worsened stroke outcomes with hypertension comorbidity are insensitive to blood pressure-lowering therapies. In an experimental stroke model with comorbid hypertension, we investigated causal roles of ang II (angiotensin II)–mediated stimulation of the brain WNK (with no lysine [K] kinases)-SPAK (STE20/SPS1-related proline/alanine-rich kinase)-NKCC1 (Na-K-Cl cotransporter) complex in worsened outcomes.
.
.
. Methods:
. Saline- or ang II–infused C57BL/6J male mice underwent stroke induced by permanent occlusion of the distal branches of the middle cerebral artery. Mice were randomly assigned to receive either vehicle dimethyl sulfoxide/PBS (2 mL/kg body weight/day, IP), a novel SPAK inhibitor, 5-chloro-N-(5-chloro-4-((4-chlorophenyl)(cyano)methyl)-2-methylphenyl)-2-hydroxybenzamide (ZT-1a‚ 5 mg/kg per day, IP) or a NF-κB (nuclear factor-κB) inhibitor TAT-NBD (transactivator of transcription-NEMO-binding domain‚ 20 mg/kg per day, IP). Activation of brain NF-κB and WNK-SPAK-NKCC1 cascade as well as ischemic stroke outcomes were examined.
.
.
. Results:
.
. Stroke triggered a 2- to 5-fold increase of WNK (isoforms 1, 2, 4), SPAK/OSR1 (oxidative stress-responsive kinase 1), and NKCC1 protein in the ang II–infused hypertensive mouse brains at 24 hours after stroke, which was associated with increased nuclear translocation of phospho-NF-κB protein in the cortical neurons (a Pearson correlation r of 0.77,
. P
. <0.005). The upregulation of WNK-SPAK-NKCC1 cascade proteins resulted from increased NF-κB recruitment on
. Wnk1, Wnk2, Wnk4, Spak
. and
. Nkcc1
. gene promoters and was attenuated by NF-κB inhibitor TAT-NBD. Poststroke administration of SPAK inhibitor ZT-1a significantly reduced WNK-SPAK-NKCC1 complex activation, brain lesion size, and neurological function deficits in the ang II–hypertensive mice without affecting blood pressure and cerebral blood flow.
.
.
.
. Conclusions:
. The ang II–induced stimulation of NF-κB transcriptional activity upregulates brain WNK-SPAK-NKCC1 cascade and contributes to worsened ischemic stroke outcomes, illustrating the brain WNK-SPAK-NKCC1 complex as a therapeutic target for stroke with comorbid hypertension.
.
Abstract.
Zhong C, Zhao H, Xie X, Qi Z, Li Y, Jia L, Zhang J, Lu Y (2022). Protein Kinase C-Mediated Hyperphosphorylation and Lateralization of Connexin 43 Are Involved in Autoimmune Myocarditis-Induced Prolongation of QRS Complex.
Frontiers in Physiology,
13Abstract:
Protein Kinase C-Mediated Hyperphosphorylation and Lateralization of Connexin 43 Are Involved in Autoimmune Myocarditis-Induced Prolongation of QRS Complex
Myocarditis is a serious and potentially life-threatening disease, which leads to cardiac dysfunction and sudden cardiac death. An increasing number of evidence suggests that myocarditis is also a malignant complication of coronavirus pneumonia, associated with heart failure and sudden cardiac death. Prolonged QRS complexes that are related to malignant arrhythmias caused by myocarditis significantly increase the risk of sudden cardiac death in patients. However, the molecular mechanisms are not fully known at present. In this study, we identify protein kinase C (PKC) as a new regulator of the QRS complex. In isolated hearts of normal rats, the PKC agonist, phorbol-12-myristate-13-acetate (PMA), induced prolongation of the QRS complex. Mechanistically, hyperphosphorylation and lateralization of connexin 43 (Cx43) by PKC induced depolymerization and internalization of Cx43 gap junction channels and prolongation of the QRS duration. Conversely, administration of the PKC inhibitor, Ro-32-0432, in experimental autoimmune myocarditis (EAM) rats after the most severe inflammation period still significantly rescued the stability of the Cx43 gap junction and alleviated prolongation of the QRS complex. Ro-32-0432 reduced phosphorylation and blocked translocation of Cx43 in EAM rat heart but did not regulate the mRNA expression level of ventricular ion channels and the other regulatory proteins, which indicates that the inhibition of PKC might have no protective effect on ion channels that generate ventricular action potential in EAM rats. These results suggest that the pharmacological inhibition of PKC ameliorates the prolongation of the QRS complex via suppression of Cx43 hyperphosphorylation, lateralization, and depolymerization of Cx43 gap junction channels in EAM rats, which provides a potential therapeutic strategy for myocarditis-induced arrhythmias.
Abstract.
Küry S, Zhang J, Besnard T, Caro-Llopis A, Zeng X, Robert SM, Josiah SS, Kiziltug E, Denommé-Pichon A-S, Cogné B, et al (2022). Rare pathogenic variants in WNK3 cause X-linked intellectual disability. Genetics in Medicine, 24(9), 1941-1951.
Wang J, Liu R, Hasan MN, Fischer S, Chen Y, Como M, Fiesler VM, Bhuiyan MIH, Dong S, Li E, et al (2022). Role of SPAK–NKCC1 signaling cascade in the choroid plexus blood–CSF barrier damage after stroke.
Journal of Neuroinflammation,
19(1).
Abstract:
Role of SPAK–NKCC1 signaling cascade in the choroid plexus blood–CSF barrier damage after stroke
Abstract
. Background
. The mechanisms underlying dysfunction of choroid plexus (ChP) blood–cerebrospinal fluid (CSF) barrier and lymphocyte invasion in neuroinflammatory responses to stroke are not well understood. In this study, we investigated whether stroke damaged the blood–CSF barrier integrity due to dysregulation of major ChP ion transport system, Na+–K+–Cl− cotransporter 1 (NKCC1), and regulatory Ste20-related proline-alanine-rich kinase (SPAK).
.
. Methods
. Sham or ischemic stroke was induced in C57Bl/6J mice. Changes on the SPAK–NKCC1 complex and tight junction proteins (TJs) in the ChP were quantified by immunofluorescence staining and immunoblotting. Immune cell infiltration in the ChP was assessed by flow cytometry and immunostaining. Cultured ChP epithelium cells (CPECs) and cortical neurons were used to evaluate H2O2-mediated oxidative stress in stimulating the SPAK–NKCC1 complex and cellular damage. In vivo or in vitro pharmacological blockade of the ChP SPAK–NKCC1 cascade with SPAK inhibitor ZT-1a or NKCC1 inhibitor bumetanide were examined.
.
. Results
. Ischemic stroke stimulated activation of the CPECs apical membrane SPAK–NKCC1 complex, NF-κB, and MMP9, which was associated with loss of the blood–CSF barrier integrity and increased immune cell infiltration into the ChP. Oxidative stress directly activated the SPAK–NKCC1 pathway and resulted in apoptosis, neurodegeneration, and NKCC1-mediated ion influx. Pharmacological blockade of the SPAK–NKCC1 pathway protected the ChP barrier integrity, attenuated ChP immune cell infiltration or neuronal death.
.
. Conclusion
. Stroke-induced pathological stimulation of the SPAK–NKCC1 cascade caused CPECs damage and disruption of TJs at the blood–CSF barrier. The ChP SPAK–NKCC1 complex emerged as a therapeutic target for attenuating ChP dysfunction and lymphocyte invasion after stroke.
.
Abstract.
Chen Z, Zhang J, Murillo-de-Ozores AR, Castañeda-Bueno M, D'Amico F, Heilig R, Manning CE, Sorrell FJ, D'Angiolella V, Fischer R, et al (2022). Sequence and structural variations determining the recruitment of WNK kinases to the KLHL3 E3 ligase.
Biochemical Journal,
479(5), 661-675.
Abstract:
Sequence and structural variations determining the recruitment of WNK kinases to the KLHL3 E3 ligase
The BTB-Kelch protein KLHL3 is a Cullin3-dependent E3 ligase that mediates the ubiquitin-dependent degradation of kinases WNK1–4 to control blood pressure and cell volume. A crystal structure of KLHL3 has defined its binding to an acidic degron motif containing a PXXP sequence that is strictly conserved in WNK1, WNK2 and WNK4. Mutations in the second proline abrograte the interaction causing the hypertension syndrome pseudohypoaldosteronism type II. WNK3 shows a diverged degron motif containing four amino acid substitutions that remove the PXXP motif raising questions as to the mechanism of its binding. To understand this atypical interaction, we determined the crystal structure of the KLHL3 Kelch domain in complex with a WNK3 peptide. The electron density enabled the complete 11-mer WNK-family degron motif to be traced for the first time revealing several conserved features not captured in previous work, including additional salt bridge and hydrogen bond interactions. Overall, the WNK3 peptide adopted a conserved binding pose except for a subtle shift to accommodate bulkier amino acid substitutions at the binding interface. At the centre, the second proline was substituted by WNK3 Thr541, providing a unique phosphorylatable residue among the WNK-family degrons. Fluorescence polarisation and structural modelling experiments revealed that its phosphorylation would abrogate the KLHL3 interaction similarly to hypertension-causing mutations. Together, these data reveal how the KLHL3 Kelch domain can accommodate the binding of multiple WNK isoforms and highlight a potential regulatory mechanism for the recruitment of WNK3.
Abstract.
Josiah SS, Meor Azlan NF, Oguro-Ando A, Zhang J (2022). Study of the Functions and Activities of Neuronal K-Cl Co-Transporter KCC2 Using Western Blotting. Journal of Visualized Experiments(190).
Seymour T, Zhang J (2021). <i>Porphyromonas Gingivalis</i> in the Pathogenesis of Alzheimer’s Disease and its Therapeutic Target. Journal of Exploratory Research in Pharmacology, 7(1), 45-53.
Zhang J, Zeng R, Labes A (2021). Editorial: Marine microbial-derived molecules and their potential medical and cosmetic applications. Frontiers in Microbiology, 12:706152
Jonniya NA, Zhang J, Kar P (2021). Molecular Mechanism of Inhibiting WNK Binding to OSR1 by Targeting the Allosteric Pocket of the OSR1-CCT Domain with Potential Antihypertensive Inhibitors: an <i>In Silico</i> Study. The Journal of Physical Chemistry B, 125(32), 9115-9129.
Bertoni A, Schaller F, Tyzio R, Gaillard S, Santini F, Xolin M, Diabira D, Vaidyanathan R, Matarazzo V, Medina I, et al (2021). Oxytocin administration in neonates shapes hippocampal circuitry and restores social behavior in a mouse model of autism.
Mol Psychiatry,
26(12), 7582-7595.
Abstract:
Oxytocin administration in neonates shapes hippocampal circuitry and restores social behavior in a mouse model of autism.
Oxytocin is an important regulator of the social brain. In some animal models of autism, notably in Magel2tm1.1Mus-deficient mice, peripheral administration of oxytocin in infancy improves social behaviors until adulthood. However, neither the mechanisms responsible for social deficits nor the mechanisms by which such oxytocin administration has long-term effects are known. Here, we aimed to clarify these oxytocin-dependent mechanisms, focusing on social memory performance. Using in situ hybridization (RNAscope), we have established that Magel2 and oxytocin receptor are co-expressed in the dentate gyrus and CA2/CA3 hippocampal regions involved in the circuitry underlying social memory. Then, we have shown that Magel2tm1.1Mus-deficient mice, evaluated in a three-chamber test, present a deficit in social memory. Next, in hippocampus, we conducted neuroanatomical and functional studies using immunostaining, oxytocin-binding experiments, ex vivo electrophysiological recordings, calcium imaging and biochemical studies. We demonstrated: an increase of the GABAergic activity of CA3-pyramidal cells associated with an increase in the quantity of oxytocin receptors and of somatostatin interneurons in both DG and CA2/CA3 regions. We also revealed a delay in the GABAergic development sequence in Magel2tm1.1Mus-deficient pups, linked to phosphorylation modifications of KCC2. Above all, we demonstrated the positive effects of subcutaneous administration of oxytocin in the mutant neonates, restoring hippocampal alterations and social memory at adulthood. Although clinical trials are debated, this study highlights the mechanisms by which peripheral oxytocin administration in neonates impacts the brain and demonstrates the therapeutic value of oxytocin to treat infants with autism spectrum disorders.
Abstract.
Author URL.
Meor Azlan NF, Koeners MP, Zhang J (2021). Regulatory control of the Na–Cl co-transporter NCC and its therapeutic potential for hypertension. Acta Pharmaceutica Sinica B, 11(5), 1117-1128.
Ostrosky-Frid M, Chávez-Canales M, Zhang J, Andrukhova O, Argaiz ER, Lerdo-De-Tejada F, Murillo-De-Ozores A, Sanchez-Navarro A, Rojas-Vega L, Bobadilla NA, et al (2021). Role of KLHL3 and dietary K+ in regulating KS-WNK1 expression.
American Journal of Physiology - Renal Physiology,
320(5), F734-F747.
Abstract:
Role of KLHL3 and dietary K+ in regulating KS-WNK1 expression
The physiological role of the shorter isoform of with no lysine kinase (WNK)1 that is exclusively expressed in the kidney (KS-WNK1), with particular abundance in the distal convoluted tubule, remains elusive. KS-WNK1, despite lacking the kinase domain, is nevertheless capable of stimulating the NaCl cotransporter, apparently through activation of WNK4. It has recently been shown that a less severe form of familial hyperkalemic hypertension featuring only hyperkalemia is caused by missense mutations in the WNK1 acidic domain that preferentially affect cullin 3 (CUL3)-Kelch-like protein 3 (KLHL3) E3-induced degradation of KS-WNK1 rather than that of full-length WNK1. Here, we show that full-length WNK1 is indeed less impacted by the CUL3-KLHL3 E3 ligase complex compared with KS-WNK1. We demonstrated that the unique 30-amino acid NH2-terminal fragment of KS-WNK1 is essential for its activating effect on the NaCl cotransporter and recognition by KLHL3. We identified specific amino acid residues in this region critical for the functional effect of KS-WNK1 and KLHL3 sensitivity. To further explore this, we generated KLHL3-R528H knockin mice that mimic human mutations causing familial hyperkalemic hypertension. These mice revealed that the KLHL3 mutation specifically increased expression of KS-WNK1 in the kidney. We also observed that in wild-type mice, the expression of KS-WNK1 was only detectable after exposure to a low-K+ diet. These findings provide new insights into the regulation and function of KS-WNK1 by the CUL3-KLHL3 complex in the distal convoluted tubule and indicate that this pathway is regulated by dietary K+ levels.
Abstract.
Salihu S, Meor Azlan NF, Josiah SS, Wu Z, Wang Y, Zhang J (2021). Role of the cation-chloride-cotransporters in the circadian system. Asian Journal of Pharmaceutical Sciences, 16(5), 589-597.
Josiah SS, Meor Azlan NF, Zhang J (2021). Targeting the WNK-SPAK/OSR1 Pathway and Cation-Chloride Cotransporters for the Therapy of Stroke.
International Journal of Molecular Sciences,
22(3), 1232-1232.
Abstract:
Targeting the WNK-SPAK/OSR1 Pathway and Cation-Chloride Cotransporters for the Therapy of Stroke
Stroke is one of the major culprits responsible for morbidity and mortality worldwide, and the currently available pharmacological strategies to combat this global disease are scanty. Cation-chloride cotransporters (CCCs) are expressed in several tissues (including neurons) and extensively contribute to the maintenance of numerous physiological functions including chloride homeostasis. Previous studies have implicated two CCCs, the Na+-K+-Cl− and K+-Cl− cotransporters (NKCCs and KCCs) in stroke episodes along with their upstream regulators, the with-no-lysine kinase (WNKs) family and STE20/SPS1-related proline/alanine rich kinase (SPAK) or oxidative stress response kinase (OSR1) via a signaling pathway. As the WNK-SPAK/OSR1 pathway reciprocally regulates NKCC and KCC, a growing body of evidence implicates over-activation and altered expression of NKCC1 in stroke pathology whilst stimulation of KCC3 during and even after a stroke event is neuroprotective. Both inhibition of NKCC1 and activation of KCC3 exert neuroprotection through reduction in intracellular chloride levels and thus could be a novel therapeutic strategy. Hence, this review summarizes the current understanding of functional regulations of the CCCs implicated in stroke with particular focus on NKCC1, KCC3, and WNK-SPAK/OSR1 signaling and discusses the current and potential pharmacological treatments for stroke.
Abstract.
Belaïdouni Y, Diabira D, Zhang J, Graziano J-C, Bader F, Montheil A, Menuet C, Wayman GA, Gaiarsa J-L (2021). The Chloride Homeostasis of CA3 Hippocampal Neurons is Not Altered in Fully Symptomatic Mepc2-null Mice.
Frontiers in Cellular Neuroscience,
15Abstract:
The Chloride Homeostasis of CA3 Hippocampal Neurons is Not Altered in Fully Symptomatic Mepc2-null Mice
Rett syndrome (RTT) is an X-linked neurodevelopmental disorder caused mainly by mutations in the MECP2 gene. Mouse models of RTT show reduced expression of the cation-chloride cotransporter KCC2 and altered chloride homeostasis at presymptomatic stages. However, whether these alterations persist to late symptomatic stages has not been studied. Here we assess KCC2 and NKCC1 expressions and chloride homeostasis in the hippocampus of early [postnatal (P) day 30–35] and late (P50–60) symptomatic male Mecp2-null (Mecp2–/y) mice. We found (i) no difference in the relative amount, but an over-phosphorylation, of KCC2 and NKCC1 between wild-type (WT) and Mecp2–/y hippocampi and (ii) no difference in the inhibitory strength, nor reversal potential, of GABAA-receptor-mediated responses in Mecp2–/y CA3 pyramidal neurons compared to WT at any stages studied. Altogether, these data indicate the presence of a functional chloride extrusion mechanism in Mecp2–/y CA3 pyramidal neurons at symptomatic stages.
Abstract.
Kahle KT, Reeves B, Karimy JK, Zhang J, Schiff SJ, Limbrick DD, Alper S, JM S (2020). A Shared, Targetable Inflammatory Mechanism Drives Hemorrhagic and Infectious Hydrocephalus. Neurosurgery, 67 (Supplement_1), nyaa447_264-nyaa447_264.
Zhang J, Bhuiyan MIH, Zhang T, Karimy J, Wu Z, Pigott VM, Zhang J, Huang H, Hassan MN, Skrzypiec AE, et al (2020). Modulation of brain cation-Cl‾ cotransport via the SPAK kinase inhibitor ZT-1a. Nature Communications, 7, 78-78.
Robert SM, Reeves BC, Alper SL, Zhang J, Kahle KT (2020). New drugs on the horizon for cerebral edema: what’s in the clinical development pipeline?. Expert Opinion on Investigational Drugs, 29(10), 1099-1105.
Meor Azlan NF, Zhang J (2020). Role of the Cation-chloride-cotransporters in Cardiovascular Disease.
Cells,
9(10).
Abstract:
Role of the Cation-chloride-cotransporters in Cardiovascular Disease
The SLC12 family of cation-chloride-cotransporters (CCCs), comprising potassium chloride cotransporters (KCCs)-mediated Cl- extrusion relative to sodium chloride cotransporters (NKCCs)-mediated Cl- loading, play vital roles in cell volume regulation and ion homeostasis. These functions of the CCCs influence a variety of physiological processes, many of which overlap with the pathophysiology of cardiovascular disease. Although not all of the cotransporters have been linked to Mendelian genetic disorders, recent studies have provided new insights into their functional role in vascular and renal cells along with their contribution to cardiovascular diseases. Particularly, an imbalance in potassium levels promote the pathogenesis of atherosclerosis and disturbances in sodium homeostasis are one of the causes of hypertension. Recent findings even suggest hypothalamic signalling as a key signalling pathway in the pathophysiology of hypertension. In this review, we summarize and discuss the role of CCCs in cardiovascular disease with particular emphasis on knowledge gained in recent years on NKCCs and KCCs.
Abstract.
Delmotte Q, Hamze M, Medina I, Buhler E, Zhang J, Belgacem YH, Porcher C (2020). Smoothened receptor signaling regulates the developmental shift of GABA polarity in rat somatosensory cortex.
J Cell Sci,
133(20).
Abstract:
Smoothened receptor signaling regulates the developmental shift of GABA polarity in rat somatosensory cortex.
Sonic hedgehog (Shh) and its patched-smoothened receptor complex control a variety of functions in the developing central nervous system, such as neural cell proliferation and differentiation. Recently, Shh signaling components have been found to be expressed at the synaptic level in the postnatal brain, suggesting a potential role in the regulation of synaptic transmission. Using in utero electroporation of constitutively active and negative-phenotype forms of the Shh signal transducer smoothened (Smo), we studied the role of Smo signaling in the development and maturation of GABAergic transmission in the somatosensory cortex. Our results show that enhancing Smo activity during development accelerates the shift from depolarizing to hyperpolarizing GABA in a manner dependent on functional expression of potassium-chloride cotransporter type 2 (KCC2, also known as SLC12A5). On the other hand, blocking Smo activity maintains the GABA response in a depolarizing state in mature cortical neurons, resulting in altered chloride homeostasis and increased seizure susceptibility. This study reveals unexpected functions of Smo signaling in the regulation of chloride homeostasis, through control of KCC2 cell-surface stability, and the timing of the GABA excitatory-to-inhibitory shift in brain maturation.
Abstract.
Author URL.
Zhang J, Cordshagen A, Medina I, Nothwang HG, Wisniewski JR, Winklhofer M, Hartmann A-M (2020). Staurosporine and NEM mainly impair WNK-SPAK/OSR1 mediated phosphorylation of KCC2 and NKCC1. PLOS ONE, 15(5), e0232967-e0232967.
Andrews K, Josiah S, Zhang J (2020). The Therapeutic Potential of Neuronal K-Cl Co-Transporter KCC2 in Huntington’s Disease and its Comorbidities. International Journal of Molecular Sciences, 21, 9142-9142.
Brown A, Meor Azlan NF, Wu Z, Zhang J (2020). WNK-SPAK/OSR1-NCC kinase signaling pathway as a novel target for the treatment of salt-sensitive hypertension. Acta Pharmacologica Sinica, 42(4), 508-517.
Brown A, Farah Meor Azlan N, Wu Z, Zhang J (2020). WNK-SPAK/OSR1-NCC kinase signaling pathway as a novel target for the treatment of salt-sensitive hypertension. Zhongguo yao li xue bao = Acta pharmacologica Sinica, 42, 508-517.
Tillman L, Zhang J (2019). Crossing the Chloride Channel: the Current and Potential Therapeutic Value of the Neuronal K+-Cl- Cotransporter KCC2. BioMed Research International, 2019
Watanabe M, Zhang J, Mansuri MS, Duan J, Karimy JK, Delpire E, Alper SL, Lifton RP, Fukuda A, Kahle KT, et al (2019). Developmentally regulated KCC2 phosphorylation is essential for dynamic GABA-mediated inhibition and survival. Science Signaling, 12(603) aaw9315
Pisella LI, Gaiarsa J-L, Diabira D, Zhang J, Khalilov I, Duan J, Kahle KT, Medina I (2019). Impaired regulation of KCC2 phosphorylation leads to neuronal network dysfunction and neurodevelopmental pathology.
Science Signaling,
12(603).
Abstract:
Impaired regulation of KCC2 phosphorylation leads to neuronal network dysfunction and neurodevelopmental pathology
Heterozygous knockin mice reveal role of the neuronal channel KCC2 in social and cognitive development.
Abstract.
Zhang J (2019). LRRK2 Signalling Pathways in Parkinson’s Disease. Archives in Neurology & Neuroscience, 2(3).
Zhang J (2019). Meet Our Editorial Board Member. Current Signal Transduction Therapy, 14(1), 1-2.
Duran D, Zeng X, Jin SC, Choi J, Nelson-Williams C, Yatsula B, Gaillard J, Furey CG, Lu Q, Timberlake AT, et al (2019). Mutations in Chromatin Modifier and Ephrin Signaling Genes in Vein of Galen Malformation.
Neuron,
101(3), 429-443.e4.
Abstract:
Mutations in Chromatin Modifier and Ephrin Signaling Genes in Vein of Galen Malformation
© 2018 Elsevier Inc. Normal vascular development includes the formation and specification of arteries, veins, and intervening capillaries. Vein of Galen malformations (VOGMs) are among the most common and severe neonatal brain arterio-venous malformations, shunting arterial blood into the brain's deep venous system through aberrant direct connections. Exome sequencing of 55 VOGM probands, including 52 parent-offspring trios, revealed enrichment of rare damaging de novo mutations in chromatin modifier genes that play essential roles in brain and vascular development. Other VOGM probands harbored rare inherited damaging mutations in Ephrin signaling genes, including a genome-wide significant mutation burden in EPHB4. Inherited mutations showed incomplete penetrance and variable expressivity, with mutation carriers often exhibiting cutaneous vascular abnormalities, suggesting a two-hit mechanism. The identified mutations collectively account for ∼30% of studied VOGM cases. These findings provide insight into disease biology and may have clinical implications for risk assessment.
Abstract.
Huang H, Song S, Banerjee S, Jiang T, Zhang J, Kahle KT, Sun D, Zhang Z (2019). The WNK-SPAK/OSR1 kinases and the cation-chloride cotransporters as therapeutic targets for neurological diseases. Aging and Disease, 10
Bhuiyan MIH, Huang H, Zhang T, Molyneaux BJ, Poloyac SM, Zhang J, Deng X, Sun D, Sun D (2018). Abstract P198: WNK-SPAK-NKCC1 Cascade Activation Contributes to Worsened Brain Damage in Mice with Hypertension Co-Morbidity after Ischemic Stroke.
Hypertension,
72(Suppl_1).
Abstract:
Abstract P198: WNK-SPAK-NKCC1 Cascade Activation Contributes to Worsened Brain Damage in Mice with Hypertension Co-Morbidity after Ischemic Stroke
. Objectives:
. The WNK-SPAK/OSR1 kinase complex plays an important role in renal salt handling and pathogenesis of hypertension by regulating ion transporters and channels. Hypertension is the most common risk factor for stroke and stroke patients with hypertension comorbidity have worsened outcome with an increased risk of dependency or death. However, the mechanisms underlying the worsened ischemic stroke pathophysiology with hypertension comorbidity remain poorly defined. In this study, we investigated roles of the WNK-SPAK-NKCC1 signaling pathway in ischemic brain damage in mice with hypertension comorbidity.
.
.
. Methods:
. Hypertension was induced in C57BL/6j male mouse (12-15 weeks) by subcutaneous infusion of 1000 ng/kg/min angiotensin II (AngII, mini-osmotic pump) for two weeks. Permanent ischemic stroke was induced by permanent occlusion of the distal branches of the left middle cerebral artery (pd-MCAO). Brain tissues were harvested for immunoblot assessment of expression levels of NKCC1, SPAK/OSR1 or WNK1-4. Infarct volume and hemisphere swelling were determined by TTC staining, and behavioral deficits were analyzed by foot fault test, cylinder test and adhesive tape removal test.
.
.
. Results:
. pd-MCAO stimulated expression of WNK proteins (isoforms 1, 2, 4), total and phosphorylated SPAK/OSR1 and NKCC1 proteins in ischemic brains of the AngII-infused hypertensive mice compared to normotensive saline controls. In parallel with the increased activation of WNK-SPAK-NKCC1 signaling, hypertensive mice displayed significantly larger infarct volume and hemispheric swelling at 24 h after pd-MCAO compared to normotensive controls. Moreover, hypertensive mice exhibited a slow recovery of neurological function after ischemic stroke compared to normotensive counterparts as assessed by sensory-motor sensitive tests.
.
.
. Conclusions:
. These results suggest that activation of the WNK-SPAK-NKCC1 complex in hypertensive ischemic brains associates, at least in part, with the worsened brain damage and neurological deficits. Pharmacological inhibition of WNK-SPAK complex has therapeutic potentials for stroke therapy with hypertension comorbidity.
.
Abstract.
Wang J, Erazo T, Ferguson FM, Buckley DL, Gomez N, Muñoz-Guardiola P, Diéguez-Martínez N, Deng X, Hao M, Massefski W, et al (2018). Structural and atropisomeric factors governing the selectivity of pyrimido-benzodiazipinones as inhibitors of kinases and bromodomains.
ACS Chemical BiologyAbstract:
Structural and atropisomeric factors governing the selectivity of pyrimido-benzodiazipinones as inhibitors of kinases and bromodomains
© 2018 American Chemical Society. Bromodomains have been pursued intensively over the past several years as emerging targets for the devel-opment of anti-cancer and anti-inflammatory agents. It has recently been shown that some kinase inhibitors are able to potently inhibit the bromodomains of BRD4. The clinical activities of PLK inhibitor BI-2536 and JAK2-FLT3 inhibitor TG101348 have been attributed to this unexpected poly-pharmacology, indicating that dual-kinase/bromodomain activity may be advantageous in a therapeutic context. However, for target validation and biological investigation, a more selec-tive target profile is desired. Here we report that benzo[e]pyrimido-[5,4-b]diazepine-6(11H)-ones, versatile ATP-site di-rected kinase pharmacophores utilized in the development of inhibitors of multiple kinases including a number of previ-ously reported kinase chemical probes, are also capable of exhibiting potent BRD4-dependent pharmacology. Using a dual kinase-bromodomain inhibitor of the kinase domains of ERK5 and LRRK2, and the bromodomain of BRD4 as a case study, we define the structure-activity relationships required to achieve dual kinase/BRD4 activity as well as how to di-rect selectivity towards inhibition of either ERK5 or BRD4. This effort resulted in identification of one of the first report-ed kinase-selective chemical probes for ERK5 (JWG-071), a BET selective inhibitor with 1 μM BRD4 IC50 (JWG-115), and additional inhibitors with rationally designed polypharmacology (JWG-047, JWG-069). Co-crystallography of seven representative inhibitors with the first bromodomain of BRD4 demonstrate that distinct atropisomeric conformers rec-ognize the kinase ATP-site and the BRD4 acetyl lysine binding site, conformational preferences supported by rigid dock-ing studies.
Abstract.
Dumon C, Diabira D, Chudotvorova I, Bader F, Sahin S, Zhang J, Porcher C, Wayman G, Medina I, Gaiarsa J-L, et al (2018). The adipocyte hormone leptin sets the emergence of hippocampal inhibition in mice. eLife, 7, e36726-e36726.
Karimy JK, Zhang J, Kurland DB, Theriault BC, Duran D, Furey CG, Gerzanich V, Simard JM, Kahle KT (2017). 166 TLR-4-Regulated Cerebrospinal Fluid Hypersecretion in Post-Hemorrhagic Hydrocephalus. Neurosurgery, 64(CN_suppl_1), 242-242.
Zhang J, Burgess JG (2017). Enhanced eicosapentaenoic acid production by a new deep-sea marine bacterium Shewanella electrodiphila MAR441(T).
PLOS ONE,
12(11).
Author URL.
Heubl M, Zhang J, Pressey JC, Al Awabdh S, Renner M, Gomez-Castro F, Moutkine I, Eugène E, Russeau M, Kahle KT, et al (2017). GABAA receptor dependent synaptic inhibition rapidly tunes KCC2 activity via the Cl--sensitive WNK1 kinase.
Nat Commun,
8(1).
Abstract:
GABAA receptor dependent synaptic inhibition rapidly tunes KCC2 activity via the Cl--sensitive WNK1 kinase.
The K+-Cl- co-transporter KCC2 (SLC12A5) tunes the efficacy of GABAA receptor-mediated transmission by regulating the intraneuronal chloride concentration [Cl-]i. KCC2 undergoes activity-dependent regulation in both physiological and pathological conditions. The regulation of KCC2 by synaptic excitation is well documented; however, whether the transporter is regulated by synaptic inhibition is unknown. Here we report a mechanism of KCC2 regulation by GABAA receptor (GABAAR)-mediated transmission in mature hippocampal neurons. Enhancing GABAAR-mediated inhibition confines KCC2 to the plasma membrane, while antagonizing inhibition reduces KCC2 surface expression by increasing the lateral diffusion and endocytosis of the transporter. This mechanism utilizes Cl- as an intracellular secondary messenger and is dependent on phosphorylation of KCC2 at threonines 906 and 1007 by the Cl--sensing kinase WNK1. We propose this mechanism contributes to the homeostasis of synaptic inhibition by rapidly adjusting neuronal [Cl-]i to GABAAR activity.
Abstract.
Author URL.
Karimy JK, Zhang J, Kurland DB, Theriault BC, Duran D, Stokum JA, Furey CG, Zhou X, Mansuri MS, Montejo J, et al (2017). Inflammation-dependent cerebrospinal fluid hypersecretion by the choroid plexus epithelium in posthemorrhagic hydrocephalus.
Nature Medicine,
23(8), 997-1003.
Abstract:
Inflammation-dependent cerebrospinal fluid hypersecretion by the choroid plexus epithelium in posthemorrhagic hydrocephalus
© 2017 Nature America, Inc. part of Springer Nature. All rights reserved. The choroid plexus epithelium (CPE) secretes higher volumes of fluid (cerebrospinal fluid, CSF) than any other epithelium and simultaneously functions as the blood-CSF barrier to gate immune cell entry into the central nervous system. Posthemorrhagic hydrocephalus (PHH), an expansion of the cerebral ventricles due to CSF accumulation following intraventricular hemorrhage (IVH), is a common disease usually treated by suboptimal CSF shunting techniques. PHH is classically attributed to primary impairments in CSF reabsorption, but little experimental evidence supports this concept. In contrast, the potential contribution of CSF secretion to PHH has received little attention. In a rat model of PHH, we demonstrate that IVH causes a Toll-like receptor 4 (TLR4)-and NF-κ B-dependent inflammatory response in the CPE that is associated with a 3-fold increase in bumetanide-sensitive CSF secretion. IVH-induced hypersecretion of CSF is mediated by TLR4-dependent activation of the Ste20-type stress kinase SPAK, which binds, phosphorylates, and stimulates the NKCC1 co-transporter at the CPE apical membrane. Genetic depletion of TLR4 or SPAK normalizes hyperactive CSF secretion rates and reduces PHH symptoms, as does treatment with drugs that antagonize TLR4-NF-κ B signaling or the SPAK-NKCC1 co-transporter complex. These data uncover a previously unrecognized contribution of CSF hypersecretion to the pathogenesis of PHH, demonstrate a new role for TLRs in regulation of the internal brain milieu, and identify a kinase-regulated mechanism of CSF secretion that could be targeted by repurposed US Food and Drug Administration (FDA)-approved drugs to treat hydrocephalus.
Abstract.
Zhang J, Karimy JK, Delpire E, Kahle KT (2017). Pharmacological targeting of SPAK kinase in disorders of impaired epithelial transport.
Expert Opinion on Therapeutic Targets,
21(8), 795-804.
Abstract:
Pharmacological targeting of SPAK kinase in disorders of impaired epithelial transport
© 2017 Informa UK Limited, trading as Taylor. &. Francis Group. Introduction: the mammalian SPS1-related proline/alanine-rich serine-threonine kinase SPAK (STK39) modulates ion transport across and between epithelial cells in response to environmental stimuli such osmotic stress and inflammation. Research over the last decade has established a central role for SPAK in the regulation of ion and water transport in the distal nephron, colonic crypts, and pancreatic ducts, and has implicated deregulated SPAK signaling in NaCl-sensitive hypertension, ulcerative colitis and Crohn’s disease, and cystic fibrosis. Areas covered: We review recent advances in our understanding of the role of SPAK kinase in the regulation of epithelial transport. We highlight how SPAK signaling–including its upstream Cl–sensitive activators, the WNK kinases, and its downstream ion transport targets, the cation- Cl–cotransporters contribute to human disease. We discuss prospects for the pharmacotherapeutic targeting of SPAK kinase in specific human disorders that feature impaired epithelial homeostasis. Expert opinion: the development of novel drugs that antagonize the SPAK-WNK interaction, inhibit SPAK kinase activity, or disrupt SPAK kinase activation by interfering with its binding to MO25α/β could be useful adjuncts in essential hypertension, inflammatory colitis, and cystic fibrosis.
Abstract.
Shekarabi M, Zhang J, Khanna AR, Ellison DH, Delpire E, Kahle KT (2017). WNK Kinase Signaling in Ion Homeostasis and Human Disease. Cell Metabolism, 25(2), 285-299.
Andrukhova O, Zhang J, Alessi D, Erben R (2016). Bone loss in KLHL3 knock-in mice characterized by a pseudohypoaldosteronism type II-like phenotype is mediated by renal PTH resistance. Bone Abstracts
Zhang J, Gao G, Begum G, Wang J, Khanna AR, Shmukler BE, Daubner GM, de los Heros P, Davies P, Varghese J, et al (2016). Functional kinomics establishes a critical node of volume-sensitive cation-Cl− cotransporter regulation in the mammalian brain.
Scientific Reports,
6(1).
Abstract:
Functional kinomics establishes a critical node of volume-sensitive cation-Cl− cotransporter regulation in the mammalian brain
AbstractCell volume homeostasis requires the dynamically regulated transport of ions across the plasmalemma. While the ensemble of ion transport proteins involved in cell volume regulation is well established, the molecular coordinators of their activities remain poorly characterized. We utilized a functional kinomics approach including a kinome-wide siRNA-phosphoproteomic screen, a high-content kinase inhibitor screen, and a kinase trapping-Orbitrap mass spectroscopy screen to systematically identify essential kinase regulators of KCC3 Thr991/Thr1048 phosphorylation – a key signaling event in cell swelling-induced regulatory volume decrease (RVD). In the mammalian brain, we found the Cl−-sensitive WNK3-SPAK kinase complex, required for cell shrinkage-induced regulatory volume decrease (RVI) via the stimulatory phosphorylation of NKCC1 (Thr203/Thr207/Thr212), is also essential for the inhibitory phosphorylation of KCC3 (Thr991/Thr1048). This is mediated in vivo by an interaction between the CCT domain in SPAK and RFXV/I domains in WNK3 and NKCC1/KCC3. Accordingly, genetic or pharmacologic WNK3-SPAK inhibition prevents cell swelling in response to osmotic stress and ameliorates post-ischemic brain swelling through a simultaneous inhibition of NKCC1-mediated Cl− uptake and stimulation of KCC3-mediated Cl− extrusion. We conclude that WNK3-SPAK is an integral component of the long-sought “Cl−/volume-sensitive kinase” of the cation-Cl− cotransporters, and functions as a molecular rheostat of cell volume in the mammalian brain.
Abstract.
Kahle KT, Schmouth J-F, Lavastre V, Latremoliere A, Zhang J, Andrews N, Omura T, Laganière J, Rochefort D, Hince P, et al (2016). Inhibition of the kinase WNK1/HSN2 ameliorates neuropathic pain by restoring GABA inhibition.
Science Signaling,
9(421).
Abstract:
Inhibition of the kinase WNK1/HSN2 ameliorates neuropathic pain by restoring GABA inhibition
Mice lacking the HSN2 form of the kinase WNK1 are protected from neuropathic pain due to nerve injury.
Abstract.
Zhang J, Deng X, Kahle KT (2016). Leveraging unique structural characteristics of WNK kinases to achieve therapeutic inhibition.
Science Signaling,
9(450).
Abstract:
Leveraging unique structural characteristics of WNK kinases to achieve therapeutic inhibition
Exploiting unique characteristics of the WNK-SPAK pathway may yield useful antihypertensive medications.
Abstract.
Emami K, Nelson A, Hack E, Zhang J, Green DH, Caldwell GS, Mesbahi E (2016). MALDI-TOF Mass Spectrometry Discriminates Known Species and Marine Environmental Isolates of Pseudoalteromonas. Frontiers in Microbiology, 7
Kahle KT, Flores B, Bharucha-Goebel D, Zhang J, Donkervoort S, Hegde M, Begum G, Duran D, Liang B, Sun D, et al (2016). Peripheral motor neuropathy is associated with defective kinase regulation of the KCC3 cotransporter.
Science Signaling,
9(439).
Abstract:
Peripheral motor neuropathy is associated with defective kinase regulation of the KCC3 cotransporter
Evaluation of a patient with peripheral motor weakness reveals a key role for phosphorylation-dependent regulation of the transporter KCC3 in the peripheral nervous system.
Abstract.
Kazlauskaite A, Martínez‐Torres RJ, Wilkie S, Kumar A, Peltier J, Gonzalez A, Johnson C, Zhang J, Hope AG, Peggie M, et al (2015). Binding to serine 65‐phosphorylated ubiquitin primes Parkin for optimal. <scp>PINK</scp>. 1‐dependent phosphorylation and activation. EMBO reports, 16(8), 939-954.
Schumacher F, Siew K, Zhang J, Johnson C, Wood N, Cleary SE, Al Maskari RS, Ferryman JT, Hardege I, Yasmin, et al (2015). Characterisation of the Cullin‐3 mutation that causes a severe form of familial hypertension and hyperkalaemia. EMBO Molecular Medicine, 7(10), 1285-1306.
Zhang J, Siew K, Macartney T, O'Shaughnessy KM, Alessi DR (2015). Critical role of the SPAK protein kinase CCT domain in controlling blood pressure.
Hum Mol Genet,
24(16), 4545-4558.
Abstract:
Critical role of the SPAK protein kinase CCT domain in controlling blood pressure.
The STE20/SPS1-related proline/alanine-rich kinase (SPAK) controls blood pressure (BP) by phosphorylating and stimulating the Na-Cl (NCC) and Na-K-2Cl (NKCC2) co-transporters, which regulate salt reabsorption in the kidney. SPAK possesses a conserved carboxy-terminal (CCT) domain, which recognises RFXV/I motifs present in its upstream activator [isoforms of the With-No-lysine (K) kinases (WNKs)] as well as its substrates (NCC and NKCC2). To define the physiological importance of the CCT domain, we generated knock-in mice in which the critical CCT domain Leu502 residue required for high affinity recognition of the RFXI/V motif was mutated to Alanine. The SPAK CCT domain defective knock-in animals are viable, and the Leu502Ala mutation abolished co-immunoprecipitation of SPAK with WNK1, NCC and NKCC2. The CCT domain defective animals displayed markedly reduced SPAK activity and phosphorylation of NCC and NKCC2 co-transporters at the residues phosphorylated by SPAK. This was also accompanied by a reduction in the expression of NCC and NKCC2 protein without changes in mRNA levels. The SPAK CCT domain knock-in mice showed typical features of Gitelman Syndrome with mild hypokalaemia, hypomagnesaemia, hypocalciuria and displayed salt wasting on switching to a low-Na diet. These observations establish that the CCT domain plays a crucial role in controlling SPAK activity and BP. Our results indicate that CCT domain inhibitors would be effective at reducing BP by lowering phosphorylation as well as expression of NCC and NKCC2.
Abstract.
Author URL.
Hatcher JM, Zhang J, Choi HG, Ito G, Alessi DR, Gray NS (2015). Discovery of a Pyrrolopyrimidine (JH-II-127), a Highly Potent, Selective, and Brain Penetrant LRRK2 Inhibitor.
ACS Med Chem Lett,
6(5), 584-589.
Abstract:
Discovery of a Pyrrolopyrimidine (JH-II-127), a Highly Potent, Selective, and Brain Penetrant LRRK2 Inhibitor.
Activating mutations in leucine-rich repeat kinase 2 (LRRK2) are present in a subset of Parkinson's disease (PD) patients and may represent an attractive therapeutic target. Here we report a 2-anilino-4-methylamino-5-chloropyrrolopyrimidine, JH-II-127 (18), as a potent and selective inhibitor of both wild-type and G2019S mutant LRRK2. Compound 18 substantially inhibits Ser910 and Ser935 phosphorylation of both wild-type LRRK2 and G2019S mutant at a concentration of 0.1-0.3 μM in a variety of cell types and is capable of inhibiting Ser935 phosphorylation in mouse brain following oral delivery of doses as low as 30 mg/kg.
Abstract.
Author URL.
Zhang J, Burgess JG (2015). Shewanella electrodiphila sp. nov. a psychrotolerant bacterium isolated from Mid-Atlantic Ridge deep-sea sediments.
Int J Syst Evol Microbiol,
65(9), 2882-2889.
Abstract:
Shewanella electrodiphila sp. nov. a psychrotolerant bacterium isolated from Mid-Atlantic Ridge deep-sea sediments.
Strains MAR441(T) and MAR445 were isolated from Mid-Atlantic Ridge sediments from a depth of 2734 m, and were found to belong to the genus Shewanella. The strains were rod-shaped, pigmented, non-motile and capable of anaerobic growth either by fermentation of carbohydrates or by anaerobic respiration. The strains utilized a variety of electron acceptors, including nitrate and ferric compounds, and could utilize peptone when grown anaerobically in a two-chambered microbial fuel cell, which used carbon cloth electrodes and delivered a stable power output of ,150-200 mW m(-2). The major fatty acids were typical of the genus Shewanella, with major components C13 : 0, iso-C13 : 0, iso-C15 : 0, C16 : 0, C16 : 1ω7c, C18 : 1ω7c and C20 : 5ω3 fatty acids. The DNA G+C content of strains MAR441(T) and MAR445 was 42.4 mol%. 16S rRNA gene sequence analysis indicated that strains MAR441(T) and MAR445 were most closely related to Shewanella olleyana (sequence similarities 97.9% to the type strain). DNA-DNA hybridization demonstrated only 15.6-37.2% relatedness between strain MAR441(T) and the type strains of related species of the genus Shewanella. Phenotypic characteristics confirmed that these isolates constituted a novel species of the genus Shewanella, for which the name Shewanella electrodiphila sp. nov. is proposed; the type strain is MAR441(T) (5ATCC BAA-2408(T) = DSM 24955(T)).
Abstract.
Author URL.
Friedel P, Kahle KT, Zhang J, Hertz N, Pisella LI, Buhler E, Schaller F, Duan J, Khanna AR, Bishop PN, et al (2015). WNK1-regulated inhibitory phosphorylation of the KCC2 cotransporter maintains the depolarizing action of GABA in immature neurons.
Science Signaling,
8(383).
Abstract:
WNK1-regulated inhibitory phosphorylation of the KCC2 cotransporter maintains the depolarizing action of GABA in immature neurons
. Immature neurons need WNK1-dependent phosphorylation of KCC2 to prevent a premature switch in the Cl
. −
. gradient and the effect of GABA.
.
Abstract.
Kahle KT, Gao G, Zhang J, Latremoliere A, Andrews N, Shang Y, Alessi D, Woolf C, Elledge S, Clapham D, et al (2014). Ronald R. Tasker Young Investigator Award 165 Promoting Endogenous GABAergic Analgesia via Kinase Modulation of Neuronal Ion Plasticity. Neurosurgery, 61(Supplement 1), 214-214.
Heros P, Zhang J, Gourlay R, Campbell D, Deak M, Macartney T, Kahle K, Alessi D (2014). SPAK/OSR1 kinases directly phosphorylate the K+‐Cl‐ co‐transporters (1109.7). The FASEB Journal, 28(S1).
Alessi DR, Zhang J, Khanna A, Hochdörfer T, Shang Y, Kahle KT (2014). The WNK-SPAK/OSR1 pathway: Master regulator of cation-chloride cotransporters.
Science Signaling(334).
Abstract:
The WNK-SPAK/OSR1 pathway: Master regulator of cation-chloride cotransporters
The WNK-SPAK/OSR1 kinase complex is composed of the kinases WNK (with no lysine) and SPAK (SPS1-related proline/alanine-rich kinase) or the SPAK homolog OSR1 (oxidative stress-responsive kinase 1). The WNK family senses changes in intracellular Cl-concentration, extracellular osmolarity, and cell volume and transduces this information to sodium (Na+), potassium (K+), and chloride (Cl-) cotransporters [collectively referred to as CCCs (cation-chloride cotransporters)] and ion channels to maintain cellular and organismal homeostasis and affect cellular morphology and behavior. Several genes encoding proteins in this pathway aremutated in human disease, and the cotransporters are targets of commonly used drugs. WNKs stimulate the kinases SPAK and OSR1, which directly phosphorylate and stimulate Cl--importing, Na+-driven CCCs or inhibit the Cl--extruding, K+-driven CCCs. These coordinated and reciprocal actions on the CCCs are triggered by an interaction between RFXV/I motifs within the WNKs and CCCs and a conserved carboxyl-terminal docking domain in SPAK and OSR1. This interaction site represents a potentially druggable node that could bemore effective than targeting the cotransporters directly. In the kidney, WNK-SPAK/OSR1 inhibition decreases epithelial NaCl reabsorption and K+secretion to lower blood pressure while maintaining serum K+. In neurons, WNK-SPAK/OSR1 inhibition could facilitate Cl-extrusion and promote γ-aminobutyric acidergic (GABAergic) inhibition. Such drugs could have efficacy as K+-sparing blood pressure-lowering agents in essential hypertension, nonaddictive analgesics in neuropathic pain, and promoters of GABAergic inhibition in diseases associated with neuronal hyperactivity, such as epilepsy, spasticity, neuropathic pain, schizophrenia, and autism.
Abstract.
de Los Heros P, Alessi DR, Gourlay R, Campbell DG, Deak M, Macartney TJ, Kahle KT, Zhang J (2014). The WNK-regulated SPAK/OSR1 kinases directly phosphorylate and inhibit the K+ -Cl- co-transporters.
Biochemical Journal,
458(3), 559-573.
Abstract:
The WNK-regulated SPAK/OSR1 kinases directly phosphorylate and inhibit the K+ -Cl- co-transporters
Precise homoeostasis of the intracellular concentration of Cl- is achieved via the co-ordinated activities of the Cl- influx and efflux. We demonstrate that the WNK (WNK lysine-deficient protein kinase)-activated SPAK (SPS1-related proline/alanine-rich kinase)/OSR1 (oxidative stress-responsive kinase 1) knownto directly phosphorylate and stimulate the N[K]CCs (Na+-K+ ion co-transporters), also promote inhibition of the KCCs (K+-Cl- co-transporters) by directly phosphorylating a recently described C-terminal threonine residue conserved in allKCCisoforms [Site-2 (Thr1048)]. First, we demonstrate that SPAK and OSR1, in the presence of theMO25 regulatory subunit, robustly phosphorylates allKCC isoforms at Site-2 in vitro. Secondly, STOCK1S-50699, a WNK pathway inhibitor, suppresses SPAK/OSR1 activation and KCC3ASite-2 phosphorylationwith similar efficiency. Thirdly, in ES (embryonic stem) cells lacking SPAK/OSR1 activity, endogenous phosphorylation of KCC isoforms at Site-2 is abolished and these cells display elevated basal activity of 86Rb+ uptake that was not markedly stimulated further by hypotonic high K+ conditions, consistent with KCC3A activation. Fourthly, a tight correlation exists between SPAK/OSR1 activity and the magnitude of KCC3A Site-2 phosphorylation. Lastly, a Site-2 alanine KCC3A mutant preventing SPAK/OSR1 phosphorylation exhibits increased activity. We also observe that KCCs are directly phosphorylated by SPAK/OSR1, at a novel Site-3 (Thr5 in KCC1/KCC3 and Thr6 in KCC2/KCC4), and a previously recognized KCC3-specific residue, Site-4 (Ser96 ). These data demonstrate that the WNK-regulated SPAK/OSR1 kinases directly phosphorylate the N[K]CCs and KCCs, promoting their stimulation and inhibition respectively. Given these reciprocal actions with anticipated net effects of increasing Cl- influx, we propose that the targeting of WNK-SPAK/OSR1 with kinase inhibitors might be a novel potent strategy to enhance cellular Cl - extrusion, with potential implications for the therapeutic modulation of epithelial and neuronal ion transport in human disease states. © 2014 the Author(s).
Abstract.
Delbroek L, Van Kolen K, Steegmans L, da Cunha R, Mandemakers W, Daneels G, De Bock PJ, Zhang J, Gevaert K, De Strooper B, et al (2013). Development of an enzyme-linked immunosorbent assay for detection of cellular and in vivo LRRK2 S935 phosphorylation.
Journal of Pharmaceutical and Biomedical Analysis,
76, 49-58.
Abstract:
Development of an enzyme-linked immunosorbent assay for detection of cellular and in vivo LRRK2 S935 phosphorylation
After the discovery of kinase activating mutations in leucine-rich repeat kinase 2 (LRRK2) as associated with autosomal dominant forms of Parkinson's disease, inhibition of the kinase is being extensively explored as a disease modifying strategy. As signaling properties and substrate(s) of LRRK2 are poorly documented, autophosphorylation has been an important readout for the enzyme's activity. Western blotting using anti-phospho-S910 or S935 LRRK2 antibodies showed effectiveness in demonstrating inhibitory effects of compounds. In this communication we describe two types of enzyme-linked immunosorbent assays (ELISA) to determine LRRK2 protein levels and kinase activity. Both assays take advantage of the sensitivity of the earlier described total and pS935 antibodies for detection (Nichols et al. Biochem. J. 2010) [10]. The first assay is based on anti-GFP-based capturing of overexpressed LRRK2 and is highly suitable to show cellular effects of kinase inhibitors in a 96-well format. In the other platform anti-LRRK2-based capturing allows detection of endogenously expressed LRRK2 in rat tissue with no significant signal in tissue from LRRK2 knockout rats. Furthermore, both assays showed a significant reduction in pS935 levels on cellular and transgenic R1441C/G LRRK2. With the anti-LRRK2 ELISA we were able to detect LRRK2 phosphorylation in human peripheral blood mononuclear cells (PBMC). To conclude, we report two sensitive assays to monitor LRRK2 expression and kinase activity in samples coming from cellular and in vivo experimental settings. Both can show their value in drug screening and biomarker development but will also be useful in the elucidation of LRRK2-mediated signaling pathways. © 2012 Elsevier B.V.
Abstract.
Yao C, Johnson WM, Gao Y, Wang W, Zhang J, Deak M, Alessi. DR, Zhu X, Mieyal JJ, Roder H, et al (2013). Kinase inhibitors arrest neurodegeneration in cell and C. elegans models of LRRK2 toxicity.
Human Molecular Genetics,
22(2), 328-344.
Abstract:
Kinase inhibitors arrest neurodegeneration in cell and C. elegans models of LRRK2 toxicity
Mutations in leucine-rich repeat kinase 2 (LRRK2) are the most frequent known cause of late-onset Parkinson's disease (PD). To explore the therapeutic potential of small molecules targeting the LRRK2 kinase domain, we characterized two LRRK2 kinase inhibitors, TTT-3002 and LRRK2-IN1, for their effects against LRRK2 activity in vitro and in Caenorhabditis elegans models of LRRK2-linked neurodegeneration. TTT-3002 and LRRK2-IN1 potently inhibited in vitro kinase activity of LRRK2 wild-type and mutant proteins, attenuated phosphorylation of cellular LRRK2 and rescued neurotoxicity of mutant LRRK2 in transfected cells. To establish whether LRRK2 kinase inhibitors can mitigate pathogenesis caused by different mutations including G2019S and R1441C located within and outside of the LRRK2 kinase domain, respectively, we evaluated effects of TTT-3002 and LRRK2-IN1 against R1441C- and G2019S-induced neurodegeneration in C. elegans models. TTT-3002 and LRRK2-IN1 rescued the behavioral deficit characteristic of dopaminergic impairment in transgenic C. elegans expressing human R1441C- and G2019S-LRRK2. The inhibitors displayed nanomolar to low micromolar rescue potency when administered either pre-symptomatically or post-symptomatically, indicating both prevention and reversal of the dopaminergic deficit. The same treatments also led to long-lasting prevention and rescue of neurodegeneration. In contrast, TTT-3002 and LRRK2-IN1 were ineffective against the neurodegenerative phenotype in transgenic worms carrying the inhibitor-resistant A2016T mutation ofLRRK2, suggesting that they elicit neuroprotective effects in vivo by targeting LRRK2 specifically. Our findings indicate that the LRRK2 kinase activity is critical for neurodegeneration caused by R1441C and G2019S mutations, suggesting that kinase inhibition of LRRK2 may represent a promising therapeutic strategy for PD. © the Author 2012. Published by Oxford University Press. All rights reserved.
Abstract.
Deng X, Elkins JM, Zhang J, Yang Q, Erazo T, Gomez N, Choi HG, Wang J, Dzamko N, Lee JD, et al (2013). Structural determinants for ERK5 (MAPK7) and leucine rich repeat kinase 2 activities of benzo[e]pyrimido-[5,4-b]diazepine-6(11H)-ones.
European Journal of Medicinal Chemistry,
70, 758-767.
Abstract:
Structural determinants for ERK5 (MAPK7) and leucine rich repeat kinase 2 activities of benzo[e]pyrimido-[5,4-b]diazepine-6(11H)-ones
The benzo[e]pyrimido-[5,4-b]diazepine-6(11H)-one core was discovered as a novel ERK5 (also known as MAPK7 and BMK1) inhibitor scaffold, previously. Further structure-activity relationship studies of this scaffold led to the discovery of ERK5-IN-1 (26) as the most selective and potent ERK5 inhibitor reported to date. 26 potently inhibits ERK5 biochemically with an IC 50 of 0.162 ± 0.006 μM and in cells with a cellular EC 50 for inhibiting epidermal growth factor induced ERK5 autophosphorylation of 0.09 ± 0.03 μM. Furthermore, 26 displays excellent selectivity over other kinases with a KINOMEscan selectivity score (S10) of 0.007, and exhibits exceptional bioavailability (F%) of 90% in mice. 26 will serve as a valuable tool compound to investigate the ERK5 signaling pathway and as a starting point for developing an ERK5 directed therapeutic agent. © 2013 Elsevier Masson SAS. All rights reserved.
Abstract.
Choi HS, Zhang J, Deng X, Hatcher JM, Patricelli MP, Zhao Z, Alessi DR, Gray NS (2012). Brain penetrant LRRK2 inhibitor.
ACS Medicinal Chemistry Letters,
3(8), 658-662.
Abstract:
Brain penetrant LRRK2 inhibitor
Activating mutations in leucine-rich repeat kinase 2 (LRRK2) are present in a subset of Parkinson's disease (PD) patients and may represent an attractive therapeutic target. Here, we report that a 2-anilino-4-methylamino-5- chloropyrimidine, HG-10-102-01 (4), is a potent and selective inhibitor of wild-type LRRK2 and the G2019S mutant. Compound 4 substantially inhibits Ser910 and Ser935 phosphorylation of both wild-type LRRK2 and G2019S mutant at a concentration of 0.1-0.3 μM in cells and is the first compound reported to be capable of inhibiting Ser910 and Ser935 phosphorylation in mouse brain following intraperitoneal delivery of doses as low as 50 mg/kg. © 2012 American Chemical Society.
Abstract.
Zhang J, Deng X, Choi HG, Alessi DR, Gray NS (2012). Characterization of TAE684 as a potent LRRK2 kinase inhibitor.
Bioorganic and Medicinal Chemistry Letters,
22(5), 1864-1869.
Abstract:
Characterization of TAE684 as a potent LRRK2 kinase inhibitor
Leucine-rich repeat kinase 2 (LRRK2) is linked to Parkinson's disease and may represent an attractive therapeutic target. Here we report a 2,4-dianilino-5-chloro-pyrimidine, TAE684, a previously reported inhibitor of anaplastic lymphoma kinase (ALK), is also a potent inhibitor of LRRK2 kinase activity (IC50of 7.8 nM against wild-type LRRK2, 6.1 nM against the G2019S mutant). TAE684 substantially inhibits Ser910 and Ser935 phosphorylation of both wild-type LRRK2 and G2019S mutant at a concentration of 0.1-0.3 μM in cells and in mouse spleen and kidney, but not in brain, following oral doses of 10 mg/kg. © 2012 Elsevier Ltd. All rights reserved.
Abstract.
Zhang. J, Zhang E, Scott K, Burgess JG (2012). Enhanced electricity production by use of reconstituted artificial consortia of estuarine bacteria grown as biofilms.
Environmental Science and Technology,
46(5), 2984-2992.
Abstract:
Enhanced electricity production by use of reconstituted artificial consortia of estuarine bacteria grown as biofilms
Microbial fuel cells (MFCs) can convert organic compounds directly into electricity by catalytic oxidation, and although MFCs have attracted considerable interest, there is little information on the electricity-generating potential of artificial bacterial biofilms. We have used acetate-fed MFCs inoculated with sediment, with two-chamber bottles and carbon cloth electrodes to deliver a maximum power output of ∼175 mW•m -2 and a stable power output of ∼105 mW•m -2. Power production was by direct transfer of electrons to the anode from bacterial consortia growing on the anode, as confirmed by cyclic voltammetry (CV) and scanning electron microscopy (SEM). Twenty different species (74 strains) of bacteria were isolated from the consortium under anaerobic conditions and cultured in the laboratory, of which 34% were found to be exoelectrogens in single-species studies. Exoelectrogenesis by members of the genera Vibrio, Enterobacter, and Citrobacter and by Bacillus stratosphericus was confirmed, by use of culture-based methods, for the first time. An MFC with a natural bacterial consortium showed higher power densities than those obtained with single strains. In addition, the maximum power output could be further increased to ∼200 mW•m -2 when an artificial consortium consisting of the best 25 exoelectrogenic isolates was used, demonstrating the potential for increased performance and underlying the importance of artificial biofilms for increasing power output. © 2012 American Chemical Society.
Abstract.
Reith AD, Bamborough P, Jandu K, Andreotti D, Mensah L, Dossang P, Choi HG, Deng X, Zhang J, Alessi DR, et al (2012). GSK2578215A; a potent and highly selective 2-arylmethyloxy-5-substitutent- N-arylbenzamide LRRK2 kinase inhibitor.
Bioorganic and Medicinal Chemistry Letters,
22(17), 5625-5629.
Abstract:
GSK2578215A; a potent and highly selective 2-arylmethyloxy-5-substitutent- N-arylbenzamide LRRK2 kinase inhibitor
Leucine-rich repeat kinase 2 (LRRK2) is a promising therapeutic target for some forms of Parkinson's disease. Here we report the discovery and characterization of 2-arylmethyloxy-5-subtitutent-N-arylbenzamides with potent LRRK2 activities exemplified by GSK2578215A which exhibits biochemical IC 50s of around 10 nM against both wild-type LRRK2 and the G2019S mutant. GSK2578215A exhibits exceptionally high selectivity for LRRK2 across the kinome, substantially inhibits Ser910 and Ser935 phosphorylation of both wild-type LRRK2 and G2019S mutant at a concentration of 0.3-1.0 μM in cells and in mouse spleen and kidney, but not in brain, following intraperitoneal injection of 100 mg/kg. © 2012 Elsevier Ltd. All rights reserved.
Abstract.
Zhang J, Zeng R (2011). Molecular cloning and expression of an extracellular α-amylase gene from an Antarctic deep sea psychrotolerant Pseudomonas stutzeri strain 7193.
World Journal of Microbiology and Biotechnology,
27(4), 841-850.
Abstract:
Molecular cloning and expression of an extracellular α-amylase gene from an Antarctic deep sea psychrotolerant Pseudomonas stutzeri strain 7193
Psychrotolerant Pseudomonas stutzeri strain 7193 capable of producing an extracellular α-amylase was isolated from deep sea sediments of Prydz Bay, Antarctic. The 59678-Da protein (AmyP) was encoded by 1665-bp gene (amyP). The deduced amino acid sequence was identified with four regions, which are conserved in amylolytic enzymes and form a catalytic domain, and was predicted to be maltotetraose forming extracellular amylase by using the I-TASSER online server. Purification of AmyP amylases from both the recombinant of Escherichia coli Top 10 F′ and strain 7193 was conducted. Biochemical characterization revealed that the optimal amylase activity was observed at pH 9.0 and temperature 40°C. The enzymes were unstable at temperatures above 30°C, and only retain half of their highest activity after incubation at 60°C for 5 min. Thin-layer chromatography analysis of the products of the amylolytic reaction showed the presence of maltotetraose, maltotriose, maltose and glucose in the starch hydrolysate. © 2010 Springer Science+Business Media B.V.
Abstract.
Zhang J, Zeng R (2008). Molecular cloning and expression of a cold-adapted lipase gene from an antarctic deep sea psychrotrophic bacterium Pseudomonas sp. 7323.
Marine Biotechnology,
10(5), 612-621.
Abstract:
Molecular cloning and expression of a cold-adapted lipase gene from an antarctic deep sea psychrotrophic bacterium Pseudomonas sp. 7323
A psychrotrophic bacterium producing a cold-adapted lipase was isolated from the deep-sea sediment of Prydz Bay, Antarctic and identified as a Pseudomonas strain. Determination of the nucleotide sequence of the gene encoding a lipase from Pseudomonas sp. 7323 (lipA) revealed that LipA is composed of 617 amino acid residues with a calculated molecular weight of 64,466 Da. LipA has a GXSXG motif, which is conserved in lipases/esterases and generally contains the active-site serine. The lipase purified from the Escherichia coli transformant (rLipA) by metal-chelating chromatography exhibited the same electrophoretic mobility as did the wild-type lipase (wLipA) purified from strain 7323, and both enzymes were quite similar in physicochemical properties. The optimal temperature and pH value for the lipases activity were 30°C and 9.0, respectively. They were unstable at temperatures above 25°C and only retained half of their highest activity after incubation at 60°C for 5 min. These results indicated that the enzymes were typical alkaline cold-adapted enzymes. Both enzymes were particularly activated by Ca2+. Additionally, the enzymes hydrolyzed p-nitrophenyl caprate and tributyrin at the highest velocity among the other p-nitrophenyl esters and triglycerides. © 2008 Springer Science+Business Media, LLC.
Abstract.
Zhang J, Zeng R (2008). Purification and characterization of a cold-adapted α-amylase produced by Nocardiopsis sp. 7326 isolated from Prydz Bay, Antarctic.
Marine Biotechnology,
10(1), 75-82.
Abstract:
Purification and characterization of a cold-adapted α-amylase produced by Nocardiopsis sp. 7326 isolated from Prydz Bay, Antarctic
An actinomycete strain 7326 producing cold-adapted α-amylase was isolated from the deep sea sediment of Prydz Bay, Antarctic. It was identified as Nocardiopsis based on morphology, 16S rRNA gene sequence analysis, and physiological and biochemical characteristics. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and zymogram activity staining of purified amylase showed a single band equal to a molecular mass of about 55 kDa. The optimal activity temperature of Nocardiopsis sp. 7326 amylase was 35°C, and the activity decreased dramatically at temperatures above 45°C. The enzyme was stable between pH 5 and 10, and exhibited a maximal activity at pH 8.0. Ca2+, Mn2+, Mg2+, Cu2+, and Co2+stimulated the activity of the enzyme significantly, and Rb2+, Hg2+, and EDTA inhibited the activity. The hydrolysates of soluble starch by the enzyme were mainly glucose, maltose, and maltotriose. This is the first report on the isolation and characterization of cold-adapted amylase from Nocardiopsis sp. © 2007 Springer Science+Business Media, LLC.
Abstract.
Zhang J, Lin S, Zeng R (2007). Cloning, expression, and characterization of a cold-adapted lipase gene from an antarctic deep-sea psychorotrophic bacterium, Psychobacter sp. 7195.
Journal of Microbiology and Biotechnology,
17(4), 604-610.
Abstract:
Cloning, expression, and characterization of a cold-adapted lipase gene from an antarctic deep-sea psychorotrophic bacterium, Psychobacter sp. 7195
A psychrotrophic strain 7195 showing extracellular lipolytic activity towards tributyrin was isolated from deep-sea sediment of Prydz Bay and identified as a Psychrobacter species. By screening a genomic DNA library of Psychrobacter sp. 7195, an open reading frame of 954 by coding for a lipase gene, lipA1, was identified, cloned, and sequenced. The deduced LipA1 consisted of 317 amino acids with a molecular mass of 35,210 kDa. It had one consensus motif, G-N-S-M-G (GXSXG), containing the putative active-site serine, which was conserved in other cold-adapted lipolytic enzymes. The recombinant LipA1 was purified by column chromatography with DEAE Sepharose CL-4B, and Sephadex G-75, and preparative polyacrylamide gel electrophoresis, in sequence. The purified enzyme showed highest activity at 30°C, and was unstable at temperatures higher than 30°C, indicating that it was a typical cold-adapted enzyme. The optimal pH for activity was 9.0, and the enzyme was stable between pH 7.0-10.0 after 24 h incubation at 4°C. The addition of Ca2+and Mg2+enhanced the enzyme activity of LipA1, whereas the Cd2+, Zn2+, Co2+, Fe3+, Hg2+, Fe2+, Rb2+, and EDTA strongly inhibited the activity. The LipA1 was activated by various detergents, such as Triton X-100, Tween 80, Tween 40, Span 60, Span 40, CHAPS, and SDS, and showed better resistance towards them. Substrate specificity analysis showed that there was a preference for trimyristin and p-nitrophenyl myristate (C14acyl groups). © the Korean Society for Microbiology and Biotechnology.
Abstract.
Zhang J, Zeng R (2007). Isolation of antarctic psychrotrophilic Pseudomonas sp. 7197 and cloning of ppa gene for inorganic pyrophosphatase (PPase).
Gaojishu Tongxin/Chinese High Technology Letters,
17(10), 1067-1071.
Abstract:
Isolation of antarctic psychrotrophilic Pseudomonas sp. 7197 and cloning of ppa gene for inorganic pyrophosphatase (PPase)
A psychrotrophile strain 7197, which produces inorganic pyrophosphatase (PPase), was isolated from the deep sea sediment of Prydz Bay, Antarctic. The morphology identification and 16S rDNA sequence analysis showed that the strain belongs to genus Pseudomonas. The ppa gene for inorganic pyrophosphatase (PPase) was cloned by PCR according to the primers. Nucleotide sequence analysis revealed an open reading frame (ORF) of 531 bp encoding the PPase. The amino acid sequence deduced from the nucleotide sequence of the ppa corresponded to a protein of 176 amino acid residues with a molecular weight of 19531 kDa. The PPase showed the highest amino acid sequence identity to the PPase of Psy-chrobacter sp. 273-4 with 97%, and had 79% and 75% of identity to the PPase of Neisseria meningitidis Z2491 and Mannheimia succiniciproducens MBEL55E respectively.
Abstract.
Zhang J, Zeng R (2007). Psychrotrophic amylolytic bacteria from deep sea sediment of Prydz Bay, Antarctic: Diversity and characterization of amylases.
World Journal of Microbiology and Biotechnology,
23(11), 1551-1557.
Abstract:
Psychrotrophic amylolytic bacteria from deep sea sediment of Prydz Bay, Antarctic: Diversity and characterization of amylases
Seventeen psychrotrophic bacteria with cold-adaptive amylolytic, lipolytic or proteolytic activity were isolated from deep sea sediment of Prydz Bay, Antarctic. They were affiliated with γ-Proteobacteria (12 strains) and gram-positive bacteria (5 strains) as determined by 16S rDNA sequencing. The amylase-producing strains belonged to genus Pseudomonas, Rhodococcus, and Nocardiopsis. Two Pseudomonas strains, 7193 and 7197, which showed highest amylolytic activity were chosen for further study. The optimal temperatures for their growth and amylase-producing were between 15 and 20°C. Both of the purified amylases showed highest activity at 40°C and pH 9.0, and retained 50% activity at 5°C. The SDS-PAGE and zymogram activity staining showed that the molecular mass of strain 7193 and 7197 amylases were about 60 and 50 kDa respectively. The Pseudomonas sp. 7193 amylase hydrolyzed soluble starch into glucose, maltose, maltotriose, and maltotetraose, indicating that it had both activities of α-amylase and glucoamylase. The product hydrolyzed by Pseudomonas sp. 7197 amylase was meltotetraose. © 2007 Springer Science+Business Media B.V.
Abstract.
Zhang J, Zeng R (2006). Cloning, expression and characterization of the cold active lipase (Lip3) from metagenomic DNA of an antarctic deep sea sediment.
Progress in Biochemistry and Biophysics,
33(12), 1207-1214.
Abstract:
Cloning, expression and characterization of the cold active lipase (Lip3) from metagenomic DNA of an antarctic deep sea sediment
The metagenomic DNA was extracted from the deep sea sediment with the depth of 900m of Prydz Bay, Antarctic. A lipase gene (lip3) with the size of 948bp was cloned from the metagenomic DNA by PCR with the primers designed. The deduced Lip3 protein was composed of 315 amino acids (AA) with a molecular mass of 34.577 ku. The motifs GFGNS(GXGXS)and G-N-S-M-G(GXSXG)in the AA sequences of Lip3 were found to be conserved in other lipase. They were most conserved sequence among the serine hydrolase and were necessary for the activity. A 35 ku of Lip3 was purified by Ni-NTA chelating sepharose column from the extract of recombinant E.coli Top 10F′ cell harboring a pLLP-OmpA plasmid inserted with lip3. The purified Lip3 was most active at 25°C and kept 22% of activity at 0°C. Only 10% of activity was retained after it was incubated at 35°C for 60 min. The optimal pH value for the Lip3 activity was 8.0. The Kmvalue of the enzyme towards p-nitrophenyl palmitate increased with the increasing of assayed temperature. These results indicated that Lip3 was a typical alkaline cold active enzyme.
Abstract.
Zhang J, Lian M, Zeng R (2006). Screening, fermentation condition and enzyme characterization of multicomponent enzymes producing Pseudomonas sp. NJ197.
Chinese Journal of Applied and Environmental Biology,
12(5), 683-687.
Abstract:
Screening, fermentation condition and enzyme characterization of multicomponent enzymes producing Pseudomonas sp. NJ197
A strain NJ197, producing cold-adapted complex enzyme, was isolated from deep sea sediment of the Prydz Bay, Antarctic. The morphological identification and 16S rDNA sequence analysis showed that it belonged to genus Pseudomonas. The optimal growth temperature of the Pseudomonas sp. NJ197 was 5-15°C, indicating that it was a psychro-tolerant bacterium. The strain could use various single carbonaceous and nitrogenous substances to produce complex enzyme. Its optimal and highest temperatures for enzyme production were 20°C and 30°C, respectively. The enzyme purification was performed by ammonium sulfate fractionation and anion exchange chromatography with DEAE cellulose-52. The activities of lipase and amylase were the strongest among the complex enzymes produced by Pseudomonas sp. NJ197, and the both were sensitive to high temperature. The optimal temperature and pH value for the lipase activity were 30°C and 9.0, while those for the amylase activity were 35°C and 9.5, respectively. The results indicated that they were typical alkaline cold-adapted enzymes, whose activities were stimulated by Ca2+, Mn2+, Cu2+, Co2+and Fe3+, and inhibited by Zn2+, Hg2+, Rb2+, Cd2+and EDTA. The cold-adapted lipase showed better resistance to inactivation of 1% detergents, such as SDS and CHAPS.
Abstract.
Chapters
Zhang J (2023). Gitelman Syndrome. In Rezaei N (Ed) Genetic Syndromes: a Comprehensive Reference Guide, Switzerland AG: Springer Nature, 1-4.
Josiah S, Meor Azlan NF, Zhang J (2021). Targeting the WNK-SPAK/OSR1 pathway and Cation-Chloride Cotransporters for the therapy of stroke. In (Ed)
Neuroprotection Rescue from Neuronal Death in the Brain, MDPI (Basel, Switzerland).: Mdpi AG, 27-48.
Abstract:
Targeting the WNK-SPAK/OSR1 pathway and Cation-Chloride Cotransporters for the therapy of stroke
Abstract.
Conferences
Meor Azlan NF, Koeners MP, Zhang J (In Press). Regulatory control of the Na–Cl co-transporter NCC and its therapeutic potential for hypertension.
Abstract:
Regulatory control of the Na–Cl co-transporter NCC and its therapeutic potential for hypertension
Abstract.
Zhang H, Zhang J, Wang R, Zhang Q, Spincemaille P, Nguyen TD, Wang Y (2021). Efficient Folded Attention for 3D Medical Image Reconstruction and Segmentation.
Abstract:
Efficient Folded Attention for 3D Medical Image Reconstruction and Segmentation
Abstract.
Zhang H, Zhang J, Wang R, Zhang Q, Gauthier SA, Spincemaille P, Nguyen TD, Wang Y (2021). Geometric loss for deep multiple sclerosis lesion segmentation.
Abstract:
Geometric loss for deep multiple sclerosis lesion segmentation
Abstract.
Wang J, Fiesler VM, Begum G, Bhuiyan MIH, Dong S, Li E, Kahle KT, Zhang J, Deng X, Yin Y, et al (2021). SPAK/OSR1 Signaling as a Novel Target for Post-Stroke Oxidative Stress Brain Injury. International Stroke Conference 2021. 10th - 12th Feb 2021.
Abstract:
SPAK/OSR1 Signaling as a Novel Target for Post-Stroke Oxidative Stress Brain Injury
Abstract.
Jun W, Fiesler VM, Begum G, Bhuiyan MIH, Shuying D, Li E, Kahle KT, Jinwei Z, Xianming D, Yan Y, et al (2021). SPAK/OSR1 Signaling as a Novel Target for Post-Stroke Oxidative Stress Brain Injury.
Author URL.
Zhang J (2020). Activation of K+–Cl-–cotransporter KCC2 by inhibiting the WNK-SPAK kinase signalling as a novel therapeutic strategy for epilepsy. the 6th International Conference on Epilepsy & Treatment. 21st - 22nd Sep 2020.
Abstract:
Activation of K+–Cl-–cotransporter KCC2 by inhibiting the WNK-SPAK kinase signalling as a novel therapeutic strategy for epilepsy
Abstract.
Zhang J, Zhang H, Wang A, Zhang Q, Sabuncu M, Spincemaille P, Nguyen TD, Wang Y (2020). Extending LOUPE for K-Space Under-Sampling Pattern Optimization in Multi-coil MRI.
Abstract:
Extending LOUPE for K-Space Under-Sampling Pattern Optimization in Multi-coil MRI
Abstract.
Côme E, Zhang J, Marques X, Hadchouel J, Kahle K, Poncer J-C, Lévi S (2020). Role of the WNK-SPAK-OSR1 signaling pathway in neuronal chloride homeostasis and in epilepsy. FENS. 10th - 11th Feb 2020.
Zhang J (2019). Activation of K+–Cl-–cotransporter KCC2 by inhibiting the WNK-SPAK kinase signalling as a novel therapeutic strategy for epilepsy. 23rd International Conference on Neurology & Neurophysiology. 18th - 19th Mar 2019.
Abstract:
Activation of K+–Cl-–cotransporter KCC2 by inhibiting the WNK-SPAK kinase signalling as a novel therapeutic strategy for epilepsy
Abstract.
Zhang H, Zhang J, Zhang Q, Kim J, Zhang S, Gauthier SA, Spincemaille P, Nguyen TD, Sabuncu M, Wang Y, et al (2019). RSANet: Recurrent Slice-Wise Attention Network for Multiple Sclerosis Lesion Segmentation.
Abstract:
RSANet: Recurrent Slice-Wise Attention Network for Multiple Sclerosis Lesion Segmentation
Abstract.
Zhang J, Bhuiyan MIH, Zhang T, Karimy J, Wu Z, Kahle KT, Sun D, Deng X (2018). Restoration of brain water homeostasis and neurological function via a novel kinase-cotransporter modulator. Cell Symposia, Aging and Metabolism. 23rd - 25th Sep 2018.
Abstract:
Restoration of brain water homeostasis and neurological function via a novel kinase-cotransporter modulator
Abstract.
Bhuiyan MIH, Huang H, Zhang T, Molyneaux BJ, Poloyac SM, Zhang J, Deng X, Sun D (2018). WNK-SPAK-NKCC1 Cascade Activation Contributes to Worsened Brain Damage in Mice with Hypertension Comorbidity after Ischemic Stroke. Joint Hypertension 2018 Scientific Sessions. 6th - 9th Sep 2018.
Abstract:
WNK-SPAK-NKCC1 Cascade Activation Contributes to Worsened Brain Damage in Mice with Hypertension Comorbidity after Ischemic Stroke
Abstract.
Karimy J, Zhang J, Medzhitov R, Simard M, Kahle K (2017). Inflammation-dependent cerebrospinal fluid hypersecretion from the choroid plexus in post-hemorrhagic hydrocephalus. 42nd FEBS congress. 10th - 14th Sep 2017.
Abstract:
Inflammation-dependent cerebrospinal fluid hypersecretion from the choroid plexus in post-hemorrhagic hydrocephalus
Abstract.
Kahle K, Schmouth J-F, Lavastre V, Latremoliere A, Zhang J, Andrews N, Dion PA, Duan J, Woolf CJ, Inquimbert P, et al (2016). Antagonism of Wnk/Hsn2 kinase ameliorates neuropathic pain by reducing maladaptive KCC2 inhibitory phosphorylation after nerve injury. 84th AANS Annual Scientific Meeting. 30th Apr - 4th May 2016.
Abstract:
Antagonism of Wnk/Hsn2 kinase ameliorates neuropathic pain by reducing maladaptive KCC2 inhibitory phosphorylation after nerve injury
Abstract.
Kahle K, Schmouth J-F, Lavastre V, Latremoliere A, Zhang J, Andrews N, Dion PA, Duan J, Woolf CJ, Inquimbert P, et al (2016). Antagonism of Wnk/Hsn2 kinase ameliorates neuropathic pain by reducing maladaptive KCC2 inhibitory phosphorylation after nerve injury.
Author URL.
Siew K, Zhang J, Schumacher F, Alessi DR, Kurtz T, O'Shaughnessy KM (2015). Pulse waveform analysis reveals vascular contributions to Gordon Syndrome and Gitelman Syndrome blood pressure homeostasis. Proceedings Abstracts of the Physiological Society. 1st - 1st Jul 2015.
Abstract:
Pulse waveform analysis reveals vascular contributions to Gordon Syndrome and Gitelman Syndrome blood pressure homeostasis
Abstract.
Keith S, Jinwei Z, Dario R. A, Kevin M. O (2014). Disruption of STE20/SPS1-related Proline/Alanine-rich Kinase (SPAK) binding lowers blood pressure and recapitulates Gitelman syndrome in mice. American Society of Nephrology Kidney Week 2014 Annual Meeting.
Abstract:
Disruption of STE20/SPS1-related Proline/Alanine-rich Kinase (SPAK) binding lowers blood pressure and recapitulates Gitelman syndrome in mice
Abstract.
Kahle KT, Gen G, Zhang J, Latremoliere A, Andrews N, Shang Y, Alessi D, Woolf C, Elledge S (2014). Promoting Endogenous GABAergic Analgesia via Kinase Modulation of Neuronal Ion Plasticity. 2014 CNS ANNUAL MEETING. 1st - 1st Aug 2014.
Abstract:
Promoting Endogenous GABAergic Analgesia via Kinase Modulation of Neuronal Ion Plasticity
Abstract.
Heros PDL, Zhang J, Gourlay R, Campbell D, Deak M, Macartney T, Kahle K, Alessi D (2014). SPAK/OSR1 kinases directly phosphorylate the K+-Cl- co-transporters. Federation of American Societies for Experimental Biology. 1st - 1st Apr 2014.
Abstract:
SPAK/OSR1 kinases directly phosphorylate the K+-Cl- co-transporters
Abstract.
Publications by year
In Press
Pracucci E, Graham R, S Alberio L, Nardi G, Cozzolino O, Pillai V, Saieva L, Walsh D, Landi S, Zhang J, et al (In Press). Circadian rhythm in cortical chloride homeostasis underpins variation in network excitability. bioRxiv
Meor Azlan NF, Koeners MP, Zhang J (In Press). Regulatory control of the Na–Cl co-transporter NCC and its therapeutic potential for hypertension.
Abstract:
Regulatory control of the Na–Cl co-transporter NCC and its therapeutic potential for hypertension
Abstract.
Wang J, Liu R, Hasan MN, Fischer S, Como M, Fiesler VM, Begum G, Chen Y, Bhuiyan MIH, Dong S, et al (In Press). Role of SPAK-NKCC1 Signaling Cascade in the Choroid Plexus Blood-CSF Barrier Damage After Stroke.
Abstract:
Role of SPAK-NKCC1 Signaling Cascade in the Choroid Plexus Blood-CSF Barrier Damage After Stroke
Abstract
. Background: the mechanisms underlying dysfunction of choroid plexus (ChP) blood-cerebrospinal fluid (CSF) barrier and lymphocyte invasion in neuroinflammatory responses to stroke are not well understood. In this study, we investigated whether stroke damaged the blood-CSF barrier integrity due to dysregulation of major ChP ion transport system Na+-K+-Cl- cotransporter (NKCC1) and regulatory Ste20-related proline-alanine-rich kinase (SPAK).
Methods: Sham or ischemic stroke was induced in C57Bl/6J mice. Changes of the SPAK-NKCC1 complex and tight junction proteins (TJs) in the ChP were quantified by immunofluorescence staining and immunoblotting. Immune cell infiltration in the ChP was assessed by flow cytometry and immunostaining. Cultured ChP epithelium cells (CPECs) and cortical neurons were used to evaluate H2O2-mediated oxidative stress in stimulating the SPAK-NKCC1 complex and cellular damage. In vivo or in vitro pharmacological blockade of the ChP SPAK-NKCC1 cascade with SPAK inhibitor ZT-1a or NKCC1 inhibitor bumetanide were examined.
Results: Ischemic stroke stimulated activation of the CPECs apical membrane SPAK-NKCC1 complex, NF-κB, and MMP9, which was associated with loss of the blood-CSF barrier integrity and increased immune cell infiltration into the ChP. Oxidative stress directly activated SPAK-NKCC1 pathway and resulted in apoptosis, neurodegeneration, and NKCC1-mediated ion influx. Pharmacological blockade of the SPAK-NKCC1 pathway protected the ChP barrier integrity, attenuated ChP immune cell infiltration or neuronal death.
Conclusion: Stroke-induced pathological stimulation of the SPAK-NKCC1 cascade caused CPECs damage and disruption of TJs at the blood-CSF barrier. The ChP SPAK-NKCC1 complex emerged as a therapeutic target for attenuating ChP dysfunction and lymphocyte invasion after stroke.
Abstract.
Chen Z, Zhang J, Heilig R, Sorrell FJ, D’Angiolella V, Fischer R, Alessi DR, Bullock AN (In Press). Sequence and structural variations determining the recruitment of WNK kinases to the KLHL3 E3 ligase.
Abstract:
Sequence and structural variations determining the recruitment of WNK kinases to the KLHL3 E3 ligase
AbstractThe BTB-Kelch protein KLHL3 is a Cullin3-dependent E3 ligase that mediates the ubiquitin-dependent degradation of kinases WNK1-4 to control blood pressure and cell volume. A crystal structure of KLHL3 has defined its binding to an acidic degron motif containing a PXXP sequence that is strictly conserved in WNK1, WNK2 and WNK4. Mutations in the second proline abrograte the interaction causing the hypertension syndrome pseudohypoaldosteronism type II. WNK3 shows a diverged degron motif containing 4 amino acid substitutions that remove the PXXP motif raising questions as to the mechanism of its binding. To understand this atypical interaction, we determined the crystal structure of the KLHL3 Kelch domain in complex with a WNK3 peptide. The electron density enabled the complete 11-mer WNK-family degron motif to be traced for the first time revealing several conserved features not captured in previous work, including additional salt bridge and hydrogen bond interactions. Overall, the WNK3 peptide adopted a conserved binding pose except for a subtle shift to accommodate bulkier amino acid substitutions at the binding interface. At the centre, the second proline was substituted by WNK3 Thr541, providing a unique phosphorylatable residue among the WNK-family degrons. Fluorescence polarisation and structural modelling experiments revealed that its phosphorylation would abrogate the KLHL3 interaction similarly to hypertension-causing mutations. Together, these data reveal how the KLHL3 Kelch domain can accommodate the binding of multiple WNK isoforms and highlight a potential regulatory mechanism for the recruitment of WNK3.
Abstract.
Delmotte Q, Medina I, Hamze M, Buhler E, Zhang J, Belgacem YH, Porcher C (In Press). Smoothened receptor Signaling regulates the developmental shift of GABA polarity in rat somatosensory cortex.
Abstract:
Smoothened receptor Signaling regulates the developmental shift of GABA polarity in rat somatosensory cortex
ABSTRACTSonic Hedgehog (Shh) and its patched-smoothened receptor complex control a variety of functions in the developing central nervous system (CNS) such as neural cell proliferation and differentiation. Recently, Shh signaling components have been found to be expressed at the synaptic level in the postnatal brain, suggesting a potential role in the regulation of synaptic transmission. Usingin uteroelectroporation of constitutively active and dominant-negative forms of the Shh co-receptor smoothened (Smo), we studied the role of Smo signaling in the development and maturation of GABAergic transmission in the somatosensory cortex. Our results show that enhancing Smo activity during development accelerates the shift from depolarizing to hyperpolarizing GABA in dependence on functional expression of potassium-chloride cotransporter type 2 (KCC2). On the other hand, blocking Smo activity maintains GABA response in a depolarizing state in mature cortical neurons resulting in altered chloride homeostasis and increased seizure susceptibility. This study reveals an unexpected function of Smo signaling on the regulation of chloride homeostasis through the control of KCC2 cell surface stability and on the timing of the GABA inhibitory/excitatory shift in brain maturation.Summary statementThe smoothened receptor controls the time course of inhibitory transmission through the stability of the potassium-chloride cotransporter type 2 at the plasma membrane.
Abstract.
2023
Zhang J, Wang J, Labes A, Zeng R (2023). Editorial: Marine microbial-derived molecules and their potential medical and cosmetic applications, volume II. Frontiers in Microbiology, 14
Zhang J, Siew K, Sun D (2023). Editorial: Targeting pumps, channels and transporters for the treatments of vascular, cardiovascular and kidney diseases. Frontiers in Pharmacology, 14
Bhuiyan MIH, Fischer S, Patel SM, Oft H, Zhang T, Foley LM, Zhang J, Hitchens TK, Molyneaux BJ, Deng X, et al (2023). Efficacy of novel SPAK inhibitor ZT-1a derivatives (1c, 1d, 1g & 1h) on improving post-stroke neurological outcome and brain lesion in mice. Neurochemistry International, 162, 105441-105441.
Zhang J (2023). Gitelman Syndrome. In Rezaei N (Ed) Genetic Syndromes: a Comprehensive Reference Guide, Switzerland AG: Springer Nature, 1-4.
Zhang J (2023). Oral Paclovid Significantly Reduces Hospitalization and Mortality in Non-hospitalized Elderly Patients with COVID-19.
Journal of Modern Biology and Drug DiscoveryAbstract:
Oral Paclovid Significantly Reduces Hospitalization and Mortality in Non-hospitalized Elderly Patients with COVID-19
The development of antiviral drugs against coronavirus has proceeded at an unprecedented pace. In December 2021, the US FDA has successively approved two oral drugs against coronavirus pneumonia (COVID-19), namely PAXLOVID™ (PF-07321332) developed by Pfizer and Molnupiravir developed by Merck by targeting papain-like protease (PLpro) and RNA-dependent RNA polymerase (RdRp), respectively. PLpro and RdRp are important for the normal life cycle of coronaviruses. Inhibition of their activities could impair the synthesis of viral RNA and aid therapeutic treatments. However, little is known about the real-world effectiveness of oral antivirals against the severe acute respiratory syndrome (SARS)-CoV-2 omicron (B.1.1.529) variant. Recent studies have shown that Paxlovid can lead to a marked reduction hospitalization or death among unvaccinated outpatients with early COVID-19, whereas it was found no significant benefit for patients aged 64 and younger. In comparison, Molnupiravir did not reduce the risk of hospitalization or death after infection in high-risk groups who had been vaccinated against the coronaviruses, but only accelerated their recovery.
Abstract.
Robert SM, Reeves BC, Kiziltug E, Duy PQ, Karimy JK, Mansuri MS, Marlier A, Allington G, Greenberg ABW, DeSpenza T, et al (2023). The choroid plexus links innate immunity to CSF dysregulation in hydrocephalus. Cell, 186(4), 764-785.e21.
2022
Robert S, Reeves B, Karimy JK, Marlier A, Kiziltug E, DeSpenza T, Singh A, Allington G, Phan D, Zhang J, et al (2022). 374 Multi-omic Analysis Identifies a SPAK Kinase-regulated Ensemble of Choroid Plexus Ion Transport Proteins Relevant for Post-infectious Hydrocephalus. Neurosurgery, 68(Supplement_1), 89-89.
D’Silva E, Meor Azlan NF, Zhang J (2022). Angiotensin II Receptor Blockers in the Management of Hypertension in Preventing Cognitive Impairment and Dementia—A Systematic Review.
Pharmaceutics,
14(10), 2123-2123.
Abstract:
Angiotensin II Receptor Blockers in the Management of Hypertension in Preventing Cognitive Impairment and Dementia—A Systematic Review
Hypertension is a known risk factor for cognition-related pathologies including dementia. The National Institute of Health and Care Excellence (NICE) guidelines recommend angiotensin (Ang) II receptor blockers (ARBs) or angiotensin-converting enzyme inhibitors (ACEIs) as a first-line treatment for hypertension. Although both ARBs and ACEIs show neuroprotective effects, ACEIs show contradictory side effects; therefore, ARBs may be a more viable option. However, trials assessing the effects of ARBs on cognition are scarce and conflicting. Therefore, the aim of this review is to conduct a systematic review and synthesise data on the influence of ARBs on cognition and dementia prevention. Five databases were searched from 1992–2022 to produce 13 randomised controlled trials (RCTs) involving 26,907 patients that compared associations of ARBs against placebos or other antihypertensives on cognition or probable dementia with a minimum duration of 3 months. ARBs showed greater cognitive benefits when compared to hydrochlorothiazide (HCTZ), beta blockers (BB), and ACEIs. Our findings showed that although ARBs are superior to some antihypertensives such as ACEIs, thiazide and beta blockers, they made no difference in comparison to the placebo in all but one sample of patients. The positive effects on cognitive performances are equal to calcium channel blockers (CCBs) and lower than statin. The neuroprotective effects of ARBs are also more beneficial when ARBs are taken at the same time as a statin. Due to these inconsistencies, robust conclusions cannot be made. Future trials are warranted and, if successful, could have positive economic implications and consequently improve quality of life.
Abstract.
Zhang J (2022). Current Methods in Studying the Functions and Activities of the SLC12 Family of Cation-Chloride-Cotransporters.
Journal of Visualized Experiments (JoVE)Abstract:
Current Methods in Studying the Functions and Activities of the SLC12 Family of Cation-Chloride-Cotransporters
The SLC12 family of cation-chloride-cotransporters (CCCs) comprises of potassium chloride cotransporters (KCCs, e.g. KCC1, KCC2, KCC3 and KCC4)-mediated Cl- extrusion, and sodium potassium chloride cotransporters (N[K]CCs, NKCC1, NKCC2 and NCC)-mediated Cl- loading. The CCCs play vital roles in cell volume regulation and ion homeostasis. Gain-of-function or loss-of-function of these ion transporters can cause diseases in many tissues. However, the experimental results needed in order to understand the functions and activities of CCCs largely rely on the experimental device, platform and set-up, disease models, and data analysis etc.
Abstract.
Pracucci E, Graham R, Alberio L, Nardi G, Cozzolino O, Pasquini G, Pillai V, Saieva L, Walsh D, Landi S, et al (2022). Diurnal rhythm in cortical chloride homeostasis underpins functional changes in visual cortex excitability.
Zhang J, Yao J, Rong M (2022). Editorial: Role of Ion Channels in Pain. Frontiers in Pharmacology, 13
Chen L, Yu J, Wan L, Wu Z, Wang G, Hu Z, Ren L, Zhou J, Qian B, Zhao X, et al (2022). Furosemide prevents membrane KCC2 downregulation during convulsant stimulation in the hippocampus. IBRO Neuroscience Reports, 12, 355-365.
Bhuiyan MIH, Young CB, Jahan I, Hasan MN, Fischer S, Meor Azlan NF, Liu M, Chattopadhyay A, Huang H, Kahle KT, et al (2022). NF-κB Signaling-Mediated Activation of WNK-SPAK-NKCC1 Cascade in Worsened Stroke Outcomes of Ang II–Hypertensive Mice.
Stroke,
53(5), 1720-1734.
Abstract:
NF-κB Signaling-Mediated Activation of WNK-SPAK-NKCC1 Cascade in Worsened Stroke Outcomes of Ang II–Hypertensive Mice
. Background:
. Worsened stroke outcomes with hypertension comorbidity are insensitive to blood pressure-lowering therapies. In an experimental stroke model with comorbid hypertension, we investigated causal roles of ang II (angiotensin II)–mediated stimulation of the brain WNK (with no lysine [K] kinases)-SPAK (STE20/SPS1-related proline/alanine-rich kinase)-NKCC1 (Na-K-Cl cotransporter) complex in worsened outcomes.
.
.
. Methods:
. Saline- or ang II–infused C57BL/6J male mice underwent stroke induced by permanent occlusion of the distal branches of the middle cerebral artery. Mice were randomly assigned to receive either vehicle dimethyl sulfoxide/PBS (2 mL/kg body weight/day, IP), a novel SPAK inhibitor, 5-chloro-N-(5-chloro-4-((4-chlorophenyl)(cyano)methyl)-2-methylphenyl)-2-hydroxybenzamide (ZT-1a‚ 5 mg/kg per day, IP) or a NF-κB (nuclear factor-κB) inhibitor TAT-NBD (transactivator of transcription-NEMO-binding domain‚ 20 mg/kg per day, IP). Activation of brain NF-κB and WNK-SPAK-NKCC1 cascade as well as ischemic stroke outcomes were examined.
.
.
. Results:
.
. Stroke triggered a 2- to 5-fold increase of WNK (isoforms 1, 2, 4), SPAK/OSR1 (oxidative stress-responsive kinase 1), and NKCC1 protein in the ang II–infused hypertensive mouse brains at 24 hours after stroke, which was associated with increased nuclear translocation of phospho-NF-κB protein in the cortical neurons (a Pearson correlation r of 0.77,
. P
. <0.005). The upregulation of WNK-SPAK-NKCC1 cascade proteins resulted from increased NF-κB recruitment on
. Wnk1, Wnk2, Wnk4, Spak
. and
. Nkcc1
. gene promoters and was attenuated by NF-κB inhibitor TAT-NBD. Poststroke administration of SPAK inhibitor ZT-1a significantly reduced WNK-SPAK-NKCC1 complex activation, brain lesion size, and neurological function deficits in the ang II–hypertensive mice without affecting blood pressure and cerebral blood flow.
.
.
.
. Conclusions:
. The ang II–induced stimulation of NF-κB transcriptional activity upregulates brain WNK-SPAK-NKCC1 cascade and contributes to worsened ischemic stroke outcomes, illustrating the brain WNK-SPAK-NKCC1 complex as a therapeutic target for stroke with comorbid hypertension.
.
Abstract.
Zhong C, Zhao H, Xie X, Qi Z, Li Y, Jia L, Zhang J, Lu Y (2022). Protein Kinase C-Mediated Hyperphosphorylation and Lateralization of Connexin 43 Are Involved in Autoimmune Myocarditis-Induced Prolongation of QRS Complex.
Frontiers in Physiology,
13Abstract:
Protein Kinase C-Mediated Hyperphosphorylation and Lateralization of Connexin 43 Are Involved in Autoimmune Myocarditis-Induced Prolongation of QRS Complex
Myocarditis is a serious and potentially life-threatening disease, which leads to cardiac dysfunction and sudden cardiac death. An increasing number of evidence suggests that myocarditis is also a malignant complication of coronavirus pneumonia, associated with heart failure and sudden cardiac death. Prolonged QRS complexes that are related to malignant arrhythmias caused by myocarditis significantly increase the risk of sudden cardiac death in patients. However, the molecular mechanisms are not fully known at present. In this study, we identify protein kinase C (PKC) as a new regulator of the QRS complex. In isolated hearts of normal rats, the PKC agonist, phorbol-12-myristate-13-acetate (PMA), induced prolongation of the QRS complex. Mechanistically, hyperphosphorylation and lateralization of connexin 43 (Cx43) by PKC induced depolymerization and internalization of Cx43 gap junction channels and prolongation of the QRS duration. Conversely, administration of the PKC inhibitor, Ro-32-0432, in experimental autoimmune myocarditis (EAM) rats after the most severe inflammation period still significantly rescued the stability of the Cx43 gap junction and alleviated prolongation of the QRS complex. Ro-32-0432 reduced phosphorylation and blocked translocation of Cx43 in EAM rat heart but did not regulate the mRNA expression level of ventricular ion channels and the other regulatory proteins, which indicates that the inhibition of PKC might have no protective effect on ion channels that generate ventricular action potential in EAM rats. These results suggest that the pharmacological inhibition of PKC ameliorates the prolongation of the QRS complex via suppression of Cx43 hyperphosphorylation, lateralization, and depolymerization of Cx43 gap junction channels in EAM rats, which provides a potential therapeutic strategy for myocarditis-induced arrhythmias.
Abstract.
Küry S, Zhang J, Besnard T, Caro-Llopis A, Zeng X, Robert SM, Josiah SS, Kiziltug E, Denommé-Pichon A-S, Cogné B, et al (2022). Rare pathogenic variants in WNK3 cause X-linked intellectual disability. Genetics in Medicine, 24(9), 1941-1951.
Wang J, Liu R, Hasan MN, Fischer S, Chen Y, Como M, Fiesler VM, Bhuiyan MIH, Dong S, Li E, et al (2022). Role of SPAK–NKCC1 signaling cascade in the choroid plexus blood–CSF barrier damage after stroke.
Journal of Neuroinflammation,
19(1).
Abstract:
Role of SPAK–NKCC1 signaling cascade in the choroid plexus blood–CSF barrier damage after stroke
Abstract
. Background
. The mechanisms underlying dysfunction of choroid plexus (ChP) blood–cerebrospinal fluid (CSF) barrier and lymphocyte invasion in neuroinflammatory responses to stroke are not well understood. In this study, we investigated whether stroke damaged the blood–CSF barrier integrity due to dysregulation of major ChP ion transport system, Na+–K+–Cl− cotransporter 1 (NKCC1), and regulatory Ste20-related proline-alanine-rich kinase (SPAK).
.
. Methods
. Sham or ischemic stroke was induced in C57Bl/6J mice. Changes on the SPAK–NKCC1 complex and tight junction proteins (TJs) in the ChP were quantified by immunofluorescence staining and immunoblotting. Immune cell infiltration in the ChP was assessed by flow cytometry and immunostaining. Cultured ChP epithelium cells (CPECs) and cortical neurons were used to evaluate H2O2-mediated oxidative stress in stimulating the SPAK–NKCC1 complex and cellular damage. In vivo or in vitro pharmacological blockade of the ChP SPAK–NKCC1 cascade with SPAK inhibitor ZT-1a or NKCC1 inhibitor bumetanide were examined.
.
. Results
. Ischemic stroke stimulated activation of the CPECs apical membrane SPAK–NKCC1 complex, NF-κB, and MMP9, which was associated with loss of the blood–CSF barrier integrity and increased immune cell infiltration into the ChP. Oxidative stress directly activated the SPAK–NKCC1 pathway and resulted in apoptosis, neurodegeneration, and NKCC1-mediated ion influx. Pharmacological blockade of the SPAK–NKCC1 pathway protected the ChP barrier integrity, attenuated ChP immune cell infiltration or neuronal death.
.
. Conclusion
. Stroke-induced pathological stimulation of the SPAK–NKCC1 cascade caused CPECs damage and disruption of TJs at the blood–CSF barrier. The ChP SPAK–NKCC1 complex emerged as a therapeutic target for attenuating ChP dysfunction and lymphocyte invasion after stroke.
.
Abstract.
Chen Z, Zhang J, Murillo-de-Ozores AR, Castañeda-Bueno M, D'Amico F, Heilig R, Manning CE, Sorrell FJ, D'Angiolella V, Fischer R, et al (2022). Sequence and structural variations determining the recruitment of WNK kinases to the KLHL3 E3 ligase.
Biochemical Journal,
479(5), 661-675.
Abstract:
Sequence and structural variations determining the recruitment of WNK kinases to the KLHL3 E3 ligase
The BTB-Kelch protein KLHL3 is a Cullin3-dependent E3 ligase that mediates the ubiquitin-dependent degradation of kinases WNK1–4 to control blood pressure and cell volume. A crystal structure of KLHL3 has defined its binding to an acidic degron motif containing a PXXP sequence that is strictly conserved in WNK1, WNK2 and WNK4. Mutations in the second proline abrograte the interaction causing the hypertension syndrome pseudohypoaldosteronism type II. WNK3 shows a diverged degron motif containing four amino acid substitutions that remove the PXXP motif raising questions as to the mechanism of its binding. To understand this atypical interaction, we determined the crystal structure of the KLHL3 Kelch domain in complex with a WNK3 peptide. The electron density enabled the complete 11-mer WNK-family degron motif to be traced for the first time revealing several conserved features not captured in previous work, including additional salt bridge and hydrogen bond interactions. Overall, the WNK3 peptide adopted a conserved binding pose except for a subtle shift to accommodate bulkier amino acid substitutions at the binding interface. At the centre, the second proline was substituted by WNK3 Thr541, providing a unique phosphorylatable residue among the WNK-family degrons. Fluorescence polarisation and structural modelling experiments revealed that its phosphorylation would abrogate the KLHL3 interaction similarly to hypertension-causing mutations. Together, these data reveal how the KLHL3 Kelch domain can accommodate the binding of multiple WNK isoforms and highlight a potential regulatory mechanism for the recruitment of WNK3.
Abstract.
Josiah SS, Meor Azlan NF, Oguro-Ando A, Zhang J (2022). Study of the Functions and Activities of Neuronal K-Cl Co-Transporter KCC2 Using Western Blotting. Journal of Visualized Experiments(190).
2021
Seymour T, Zhang J (2021). <i>Porphyromonas Gingivalis</i> in the Pathogenesis of Alzheimer’s Disease and its Therapeutic Target. Journal of Exploratory Research in Pharmacology, 7(1), 45-53.
Zhang J, Labes A, Zeng R (2021). Book: Marine Microbial-Derived Molecules and Their Potential Medical and Cosmetic Applications., Lausanne: Frontiers Media SA.
Zhang J, Zeng R, Labes A (2021). Editorial: Marine microbial-derived molecules and their potential medical and cosmetic applications. Frontiers in Microbiology, 12:706152
Zhang H, Zhang J, Wang R, Zhang Q, Spincemaille P, Nguyen TD, Wang Y (2021). Efficient Folded Attention for 3D Medical Image Reconstruction and Segmentation.
Abstract:
Efficient Folded Attention for 3D Medical Image Reconstruction and Segmentation
Abstract.
Zhang H, Zhang J, Wang R, Zhang Q, Gauthier SA, Spincemaille P, Nguyen TD, Wang Y (2021). Geometric loss for deep multiple sclerosis lesion segmentation.
Abstract:
Geometric loss for deep multiple sclerosis lesion segmentation
Abstract.
Jonniya NA, Zhang J, Kar P (2021). Molecular Mechanism of Inhibiting WNK Binding to OSR1 by Targeting the Allosteric Pocket of the OSR1-CCT Domain with Potential Antihypertensive Inhibitors: an <i>In Silico</i> Study. The Journal of Physical Chemistry B, 125(32), 9115-9129.
(2021). Neuroprotection: Rescue from Neuronal Death in the Brain., MDPI.
Bertoni A, Schaller F, Tyzio R, Gaillard S, Santini F, Xolin M, Diabira D, Vaidyanathan R, Matarazzo V, Medina I, et al (2021). Oxytocin administration in neonates shapes hippocampal circuitry and restores social behavior in a mouse model of autism.
Mol Psychiatry,
26(12), 7582-7595.
Abstract:
Oxytocin administration in neonates shapes hippocampal circuitry and restores social behavior in a mouse model of autism.
Oxytocin is an important regulator of the social brain. In some animal models of autism, notably in Magel2tm1.1Mus-deficient mice, peripheral administration of oxytocin in infancy improves social behaviors until adulthood. However, neither the mechanisms responsible for social deficits nor the mechanisms by which such oxytocin administration has long-term effects are known. Here, we aimed to clarify these oxytocin-dependent mechanisms, focusing on social memory performance. Using in situ hybridization (RNAscope), we have established that Magel2 and oxytocin receptor are co-expressed in the dentate gyrus and CA2/CA3 hippocampal regions involved in the circuitry underlying social memory. Then, we have shown that Magel2tm1.1Mus-deficient mice, evaluated in a three-chamber test, present a deficit in social memory. Next, in hippocampus, we conducted neuroanatomical and functional studies using immunostaining, oxytocin-binding experiments, ex vivo electrophysiological recordings, calcium imaging and biochemical studies. We demonstrated: an increase of the GABAergic activity of CA3-pyramidal cells associated with an increase in the quantity of oxytocin receptors and of somatostatin interneurons in both DG and CA2/CA3 regions. We also revealed a delay in the GABAergic development sequence in Magel2tm1.1Mus-deficient pups, linked to phosphorylation modifications of KCC2. Above all, we demonstrated the positive effects of subcutaneous administration of oxytocin in the mutant neonates, restoring hippocampal alterations and social memory at adulthood. Although clinical trials are debated, this study highlights the mechanisms by which peripheral oxytocin administration in neonates impacts the brain and demonstrates the therapeutic value of oxytocin to treat infants with autism spectrum disorders.
Abstract.
Author URL.
Meor Azlan NF, Koeners MP, Zhang J (2021). Regulatory control of the Na–Cl co-transporter NCC and its therapeutic potential for hypertension. Acta Pharmaceutica Sinica B, 11(5), 1117-1128.
Ostrosky-Frid M, Chávez-Canales M, Zhang J, Andrukhova O, Argaiz ER, Lerdo-De-Tejada F, Murillo-De-Ozores A, Sanchez-Navarro A, Rojas-Vega L, Bobadilla NA, et al (2021). Role of KLHL3 and dietary K+ in regulating KS-WNK1 expression.
American Journal of Physiology - Renal Physiology,
320(5), F734-F747.
Abstract:
Role of KLHL3 and dietary K+ in regulating KS-WNK1 expression
The physiological role of the shorter isoform of with no lysine kinase (WNK)1 that is exclusively expressed in the kidney (KS-WNK1), with particular abundance in the distal convoluted tubule, remains elusive. KS-WNK1, despite lacking the kinase domain, is nevertheless capable of stimulating the NaCl cotransporter, apparently through activation of WNK4. It has recently been shown that a less severe form of familial hyperkalemic hypertension featuring only hyperkalemia is caused by missense mutations in the WNK1 acidic domain that preferentially affect cullin 3 (CUL3)-Kelch-like protein 3 (KLHL3) E3-induced degradation of KS-WNK1 rather than that of full-length WNK1. Here, we show that full-length WNK1 is indeed less impacted by the CUL3-KLHL3 E3 ligase complex compared with KS-WNK1. We demonstrated that the unique 30-amino acid NH2-terminal fragment of KS-WNK1 is essential for its activating effect on the NaCl cotransporter and recognition by KLHL3. We identified specific amino acid residues in this region critical for the functional effect of KS-WNK1 and KLHL3 sensitivity. To further explore this, we generated KLHL3-R528H knockin mice that mimic human mutations causing familial hyperkalemic hypertension. These mice revealed that the KLHL3 mutation specifically increased expression of KS-WNK1 in the kidney. We also observed that in wild-type mice, the expression of KS-WNK1 was only detectable after exposure to a low-K+ diet. These findings provide new insights into the regulation and function of KS-WNK1 by the CUL3-KLHL3 complex in the distal convoluted tubule and indicate that this pathway is regulated by dietary K+ levels.
Abstract.
Salihu S, Meor Azlan NF, Josiah SS, Wu Z, Wang Y, Zhang J (2021). Role of the cation-chloride-cotransporters in the circadian system. Asian Journal of Pharmaceutical Sciences, 16(5), 589-597.
SUN D, Deng X, Zhang J, Hossain Bhuiyan MI, Molyneaux BJ (2021). SPAK kinase inhibitors as neuroprotective agents.
Abstract:
SPAK kinase inhibitors as neuroprotective agents
The present disclosure is concerned with N-(5-chloro-4-((4-chlorophenyl)(cyano)methyl)-2-methylphenyl)benzamide compounds that are capable of inhibiting SPAK kinase function, methods of treating hypoxic brain injuries due to, for example, ischemic stroke. This abstract is intended as a scanning tool for purposes of searching in the particular art and is not intended to be limiting of the present invention.
Abstract.
Sun D, Deng X, Zhang J, BHUIYAN MOHAMMAD I, MOLYNEAUX B (2021). SPAK kinase inhibitors as neuroprotective agents.
Abstract:
SPAK kinase inhibitors as neuroprotective agents
The present disclosure is concerned with N-(5-chloro-4-((4-chlorophenyl)(cyano)methyl)-2-methylphenyl)benzamide compounds that are capable of inhibiting SPAK kinase function, methods of treating hypoxic brain injuries due to, for example, ischemic stroke. This abstract is intended as a scanning tool for purposes of searching in the particular art and is not intended to be limiting of the present invention.
Abstract.
Wang J, Fiesler VM, Begum G, Bhuiyan MIH, Dong S, Li E, Kahle KT, Zhang J, Deng X, Yin Y, et al (2021). SPAK/OSR1 Signaling as a Novel Target for Post-Stroke Oxidative Stress Brain Injury. International Stroke Conference 2021. 10th - 12th Feb 2021.
Abstract:
SPAK/OSR1 Signaling as a Novel Target for Post-Stroke Oxidative Stress Brain Injury
Abstract.
Jun W, Fiesler VM, Begum G, Bhuiyan MIH, Shuying D, Li E, Kahle KT, Jinwei Z, Xianming D, Yan Y, et al (2021). SPAK/OSR1 Signaling as a Novel Target for Post-Stroke Oxidative Stress Brain Injury.
Author URL.
Josiah SS, Meor Azlan NF, Zhang J (2021). Targeting the WNK-SPAK/OSR1 Pathway and Cation-Chloride Cotransporters for the Therapy of Stroke.
International Journal of Molecular Sciences,
22(3), 1232-1232.
Abstract:
Targeting the WNK-SPAK/OSR1 Pathway and Cation-Chloride Cotransporters for the Therapy of Stroke
Stroke is one of the major culprits responsible for morbidity and mortality worldwide, and the currently available pharmacological strategies to combat this global disease are scanty. Cation-chloride cotransporters (CCCs) are expressed in several tissues (including neurons) and extensively contribute to the maintenance of numerous physiological functions including chloride homeostasis. Previous studies have implicated two CCCs, the Na+-K+-Cl− and K+-Cl− cotransporters (NKCCs and KCCs) in stroke episodes along with their upstream regulators, the with-no-lysine kinase (WNKs) family and STE20/SPS1-related proline/alanine rich kinase (SPAK) or oxidative stress response kinase (OSR1) via a signaling pathway. As the WNK-SPAK/OSR1 pathway reciprocally regulates NKCC and KCC, a growing body of evidence implicates over-activation and altered expression of NKCC1 in stroke pathology whilst stimulation of KCC3 during and even after a stroke event is neuroprotective. Both inhibition of NKCC1 and activation of KCC3 exert neuroprotection through reduction in intracellular chloride levels and thus could be a novel therapeutic strategy. Hence, this review summarizes the current understanding of functional regulations of the CCCs implicated in stroke with particular focus on NKCC1, KCC3, and WNK-SPAK/OSR1 signaling and discusses the current and potential pharmacological treatments for stroke.
Abstract.
Josiah S, Meor Azlan NF, Zhang J (2021). Targeting the WNK-SPAK/OSR1 pathway and Cation-Chloride Cotransporters for the therapy of stroke. In (Ed)
Neuroprotection Rescue from Neuronal Death in the Brain, MDPI (Basel, Switzerland).: Mdpi AG, 27-48.
Abstract:
Targeting the WNK-SPAK/OSR1 pathway and Cation-Chloride Cotransporters for the therapy of stroke
Abstract.
Belaïdouni Y, Diabira D, Zhang J, Graziano J-C, Bader F, Montheil A, Menuet C, Wayman GA, Gaiarsa J-L (2021). The Chloride Homeostasis of CA3 Hippocampal Neurons is Not Altered in Fully Symptomatic Mepc2-null Mice.
Frontiers in Cellular Neuroscience,
15Abstract:
The Chloride Homeostasis of CA3 Hippocampal Neurons is Not Altered in Fully Symptomatic Mepc2-null Mice
Rett syndrome (RTT) is an X-linked neurodevelopmental disorder caused mainly by mutations in the MECP2 gene. Mouse models of RTT show reduced expression of the cation-chloride cotransporter KCC2 and altered chloride homeostasis at presymptomatic stages. However, whether these alterations persist to late symptomatic stages has not been studied. Here we assess KCC2 and NKCC1 expressions and chloride homeostasis in the hippocampus of early [postnatal (P) day 30–35] and late (P50–60) symptomatic male Mecp2-null (Mecp2–/y) mice. We found (i) no difference in the relative amount, but an over-phosphorylation, of KCC2 and NKCC1 between wild-type (WT) and Mecp2–/y hippocampi and (ii) no difference in the inhibitory strength, nor reversal potential, of GABAA-receptor-mediated responses in Mecp2–/y CA3 pyramidal neurons compared to WT at any stages studied. Altogether, these data indicate the presence of a functional chloride extrusion mechanism in Mecp2–/y CA3 pyramidal neurons at symptomatic stages.
Abstract.
2020
Kahle KT, Reeves B, Karimy JK, Zhang J, Schiff SJ, Limbrick DD, Alper S, JM S (2020). A Shared, Targetable Inflammatory Mechanism Drives Hemorrhagic and Infectious Hydrocephalus. Neurosurgery, 67 (Supplement_1), nyaa447_264-nyaa447_264.
Zhang J (2020). Activation of K+–Cl-–cotransporter KCC2 by inhibiting the WNK-SPAK kinase signalling as a novel therapeutic strategy for epilepsy. the 6th International Conference on Epilepsy & Treatment. 21st - 22nd Sep 2020.
Abstract:
Activation of K+–Cl-–cotransporter KCC2 by inhibiting the WNK-SPAK kinase signalling as a novel therapeutic strategy for epilepsy
Abstract.
Zhang J, Zhang H, Wang A, Zhang Q, Sabuncu M, Spincemaille P, Nguyen TD, Wang Y (2020). Extending LOUPE for K-Space Under-Sampling Pattern Optimization in Multi-coil MRI.
Abstract:
Extending LOUPE for K-Space Under-Sampling Pattern Optimization in Multi-coil MRI
Abstract.
Zhang J, Bhuiyan MIH, Zhang T, Karimy J, Wu Z, Pigott VM, Zhang J, Huang H, Hassan MN, Skrzypiec AE, et al (2020). Modulation of brain cation-Cl‾ cotransport via the SPAK kinase inhibitor ZT-1a. Nature Communications, 7, 78-78.
Robert SM, Reeves BC, Alper SL, Zhang J, Kahle KT (2020). New drugs on the horizon for cerebral edema: what’s in the clinical development pipeline?. Expert Opinion on Investigational Drugs, 29(10), 1099-1105.
Bertoni A, Schaller F, Tyzio R, Gaillard S, Santini F, Xolin M, Diabira D, Vaidyanathan R, Matarazzo V, Medina I, et al (2020). Oxytocin administration in neonates shapes the hippocampal circuitry and restores social behavior in a mouse model of autism.
Rong M, Zhang J, Yao J (2020). Role of Ion Channels in Pain.
Frontiers in PharmacologyAbstract:
Role of Ion Channels in Pain
Ion channels allow ions to pass through cell membrane and are essential for a host of cell functions in many organs. Due to localization in primary sensory neurons and other key structures in pain processing, ion channel is regarded as a major class of drug targets for modulating pain sensation and controlling chronic pain. Among these channels, sodium channels, calcium channels, transient receptor potential (TRP) channels, PIEZO and purinergic P2X3 channels have been reported to relieve pain. Piezo2 mediates inflammation- and nerve injury-induced sensitized mechanical pain, and suggest that targeting PIEZO2 might be an effective strategy for treating mechanical allodynia. Clinical studies of P2X3 have shown promise in treatment for bladder pain and pain associated with osteoarthritis. Further investigation of the mechanisms of ion channel that are related to nociception could be key in developing a new class of therapeutics for many diseases.
Among the Food and Drug Administration-approved drugs, 18% of drugs targeting human receptors are targeted ion channels. Ziconotide is licensed for severe, intractable, and chronic cancer and non-malignant pain. Dermal patches containing 8% capsaicin (NGX-4010) are currently approved in the European Union for various forms of peripheral neuropathic pain. This Research Topic will shed light on their diverse mechanisms of ion channels in pain, and provide new strategies of treatment.
The Research Topic will cover, but is not limited to the following:
• the fundamentals of ion channels, in terms of function, modelling, regulation, molecular biology, trafficking, structure, and pharmacology in pain.
• the agonists of ion channels associated with pain, their interaction mechanisms, their applications in treating pain.
Abstract.
Meor Azlan NF, Zhang J (2020). Role of the Cation-chloride-cotransporters in Cardiovascular Disease.
Abstract:
Role of the Cation-chloride-cotransporters in Cardiovascular Disease
The SLC12 family of cation-chloride-cotransporters (CCCs), comprising potassium chloride cotransporters (KCCs)-mediated Cl- extrusion relative to sodium chloride cotransporters (NKCCs)-mediated Cl- loading, play vital roles in cell volume regulation and ion homeostasis. These functions of the CCCs influence a variety of physiological processes, many of which overlap with the pathophysiology of cardiovascular disease. Although not all of the cotransporters have been linked to Mendelian genetic disorders, recent studies have provided new insights into their functional role in vascular and renal cells along with their contribution to cardiovascular diseases. Particularly, an imbalance in potassium levels promote the pathogenesis of atherosclerosis and disturbances in sodium homeostasis are one of the causes of hypertension. Recent findings even suggest hypothalamic signalling as a key signalling pathway in the pathophysiology of hypertension. In this review, we summarize and discuss the role of CCCs in cardiovascular disease with particular emphasis on knowledge gained in recent years on NKCCs and KCCs.
Abstract.
Meor Azlan NF, Zhang J (2020). Role of the Cation-chloride-cotransporters in Cardiovascular Disease.
Cells,
9(10).
Abstract:
Role of the Cation-chloride-cotransporters in Cardiovascular Disease
The SLC12 family of cation-chloride-cotransporters (CCCs), comprising potassium chloride cotransporters (KCCs)-mediated Cl- extrusion relative to sodium chloride cotransporters (NKCCs)-mediated Cl- loading, play vital roles in cell volume regulation and ion homeostasis. These functions of the CCCs influence a variety of physiological processes, many of which overlap with the pathophysiology of cardiovascular disease. Although not all of the cotransporters have been linked to Mendelian genetic disorders, recent studies have provided new insights into their functional role in vascular and renal cells along with their contribution to cardiovascular diseases. Particularly, an imbalance in potassium levels promote the pathogenesis of atherosclerosis and disturbances in sodium homeostasis are one of the causes of hypertension. Recent findings even suggest hypothalamic signalling as a key signalling pathway in the pathophysiology of hypertension. In this review, we summarize and discuss the role of CCCs in cardiovascular disease with particular emphasis on knowledge gained in recent years on NKCCs and KCCs.
Abstract.
Côme E, Zhang J, Marques X, Hadchouel J, Kahle K, Poncer J-C, Lévi S (2020). Role of the WNK-SPAK-OSR1 signaling pathway in neuronal chloride homeostasis and in epilepsy. FENS. 10th - 11th Feb 2020.
Delmotte Q, Hamze M, Medina I, Buhler E, Zhang J, Belgacem YH, Porcher C (2020). Smoothened receptor signaling regulates the developmental shift of GABA polarity in rat somatosensory cortex.
J Cell Sci,
133(20).
Abstract:
Smoothened receptor signaling regulates the developmental shift of GABA polarity in rat somatosensory cortex.
Sonic hedgehog (Shh) and its patched-smoothened receptor complex control a variety of functions in the developing central nervous system, such as neural cell proliferation and differentiation. Recently, Shh signaling components have been found to be expressed at the synaptic level in the postnatal brain, suggesting a potential role in the regulation of synaptic transmission. Using in utero electroporation of constitutively active and negative-phenotype forms of the Shh signal transducer smoothened (Smo), we studied the role of Smo signaling in the development and maturation of GABAergic transmission in the somatosensory cortex. Our results show that enhancing Smo activity during development accelerates the shift from depolarizing to hyperpolarizing GABA in a manner dependent on functional expression of potassium-chloride cotransporter type 2 (KCC2, also known as SLC12A5). On the other hand, blocking Smo activity maintains the GABA response in a depolarizing state in mature cortical neurons, resulting in altered chloride homeostasis and increased seizure susceptibility. This study reveals unexpected functions of Smo signaling in the regulation of chloride homeostasis, through control of KCC2 cell-surface stability, and the timing of the GABA excitatory-to-inhibitory shift in brain maturation.
Abstract.
Author URL.
Zhang J (2020). Special Collection on Targeting Ion Channels and Transporters as New Strategies of Treatments for Brain Disorders.
ASN NEUROAbstract:
Special Collection on Targeting Ion Channels and Transporters as New Strategies of Treatments for Brain Disorders
Homeostasis of the intracellular ionic milieu is essential for the proper functioning of a diverse group of cellular processes. Ion channels and transporters are transmembrane proteins that allow the passive flow of ions, both in and out of cells or cellular organelles, following their electrochemical gradients. Proper maintenance of ion channels and transporters is critical in a range of cellular activities, such as signal transduction, neurotransmitter release, nerve impulses, muscle contraction, hormone secretion, volume regulation, growth, motility, apoptosis, as well as the brain's ion and water homeostasis. Dysfunction of ion channels or
transporters can result in numerous neurological disorders including epilepsy, autism, post-surgical complication, neuropathic pain and neuropsychiatric disorders.
The Human Genome Project has identified more than 400 putative ion channels, a large fraction of which is still to be explored. 16% of all Food and Drug Administration-approved drugs target ion channels and transporters, highlighting their importance in the disease process.
This Special Issue welcomes the submission of original research and review articles that shed light on the importance of ion channels and transporters in the pathophysiology of brain diseases and suggest new strategies for treatment.
Abstract.
Zhang J (2020). Special Issue "Regulation and Control of Intracellular Signalling".
ProcessesAbstract:
Special Issue "Regulation and Control of Intracellular Signalling"
Almost all aspects of cellular processes and functions are dependent on intracellular signalling initiated at the cell surface. The response of cells to signalling molecules, e.g. growth factors, is determined by their complement of expressed receptors and pathways that transduce and transmit these signals to intracellular compartments; and the enzymes, ion channels/transporters, and cytoskeletal proteins that ultimately mediate the effects of the signalling molecules. Several primary classes of signalling systems either comprise ligand-gated ion channels or consist of receptor tyrosine kinases or utilise G protein-linked signals in a multistep process, operating at different time courses from milliseconds to minutes, providing great flexibility for intercellular communication. In most cases, the initial steps in the signalling system typically generate a second messenger inside the cell, and this second messenger then activates a number of proteins, including protein kinases that modify cellular processes. Intracellular signal transduction may target transcription factors that function to regulate gene expression and connect the cell surface to the nucleus, thus determining the differentiated and functional state of cells, or in response to extracellular stimuli. Abnormalities in these pathways cause many diseases, including hypertension, cancer, diabetes, neurodegeneration and inflammation, etc. Deciphering how disruptions in signalling networks lead to disease will reveal novel drug targets and improved strategies to treat these maladies.
This Special Issue on “Regulation and Control of Intracellular Signalling” aims to curate novel advances in deciphering intracellular signalling networks for drug discovery and disease treatment. Topics include but are not limited to:
New intracellular signalling networks that are genetically and epigenetically altered in human diseases, leading to constitutive pathway activation or suppression;
New “-omic” technologies or high-throughput methods to reveal molecular interactions in a real and quantitative way within intracellular signalling networks;
New compounds that selectively inhibit altered proteins that are critical for the maintenance of the transformed phenotype and which have shown unprecedented clinical activity in genetically defined subsets of diseases;
New computational approaches towards signal transduction pathways.
Abstract.
Zhang J, Cordshagen A, Medina I, Nothwang HG, Wisniewski JR, Winklhofer M, Hartmann A-M (2020). Staurosporine and NEM mainly impair WNK-SPAK/OSR1 mediated phosphorylation of KCC2 and NKCC1. PLOS ONE, 15(5), e0232967-e0232967.
Andrews K, Josiah S, Zhang J (2020). The Therapeutic Potential of Neuronal K-Cl Co-Transporter KCC2 in Huntington’s Disease and its Comorbidities. International Journal of Molecular Sciences, 21, 9142-9142.
Brown A, Meor Azlan NF, Wu Z, Zhang J (2020). WNK-SPAK/OSR1-NCC kinase signaling pathway as a novel target for the treatment of salt-sensitive hypertension. Acta Pharmacologica Sinica, 42(4), 508-517.
Brown A, Farah Meor Azlan N, Wu Z, Zhang J (2020). WNK-SPAK/OSR1-NCC kinase signaling pathway as a novel target for the treatment of salt-sensitive hypertension. Zhongguo yao li xue bao = Acta pharmacologica Sinica, 42, 508-517.
2019
Zhang J (2019). Activation of K+–Cl-–cotransporter KCC2 by inhibiting the WNK-SPAK kinase signalling as a novel therapeutic strategy for epilepsy. 23rd International Conference on Neurology & Neurophysiology. 18th - 19th Mar 2019.
Abstract:
Activation of K+–Cl-–cotransporter KCC2 by inhibiting the WNK-SPAK kinase signalling as a novel therapeutic strategy for epilepsy
Abstract.
Tillman L, Zhang J (2019). Crossing the Chloride Channel: the Current and Potential Therapeutic Value of the Neuronal K+-Cl- Cotransporter KCC2. BioMed Research International, 2019
Watanabe M, Zhang J, Mansuri MS, Duan J, Karimy JK, Delpire E, Alper SL, Lifton RP, Fukuda A, Kahle KT, et al (2019). Developmentally regulated KCC2 phosphorylation is essential for dynamic GABA-mediated inhibition and survival. Science Signaling, 12(603) aaw9315
Pisella LI, Gaiarsa J-L, Diabira D, Zhang J, Khalilov I, Duan J, Kahle KT, Medina I (2019). Impaired KCC2 phosphorylation leads to neuronal network dysfunction and neurodevelopmental pathogenesis.
Pisella LI, Gaiarsa J-L, Diabira D, Zhang J, Khalilov I, Duan J, Kahle KT, Medina I (2019). Impaired regulation of KCC2 phosphorylation leads to neuronal network dysfunction and neurodevelopmental pathology.
Science Signaling,
12(603).
Abstract:
Impaired regulation of KCC2 phosphorylation leads to neuronal network dysfunction and neurodevelopmental pathology
Heterozygous knockin mice reveal role of the neuronal channel KCC2 in social and cognitive development.
Abstract.
Zhang J (2019). LRRK2 Signalling Pathways in Parkinson’s Disease. Archives in Neurology & Neuroscience, 2(3).
Zhang J (2019). Meet Our Editorial Board Member. Current Signal Transduction Therapy, 14(1), 1-2.
Duran D, Zeng X, Jin SC, Choi J, Nelson-Williams C, Yatsula B, Gaillard J, Furey CG, Lu Q, Timberlake AT, et al (2019). Mutations in Chromatin Modifier and Ephrin Signaling Genes in Vein of Galen Malformation.
Neuron,
101(3), 429-443.e4.
Abstract:
Mutations in Chromatin Modifier and Ephrin Signaling Genes in Vein of Galen Malformation
© 2018 Elsevier Inc. Normal vascular development includes the formation and specification of arteries, veins, and intervening capillaries. Vein of Galen malformations (VOGMs) are among the most common and severe neonatal brain arterio-venous malformations, shunting arterial blood into the brain's deep venous system through aberrant direct connections. Exome sequencing of 55 VOGM probands, including 52 parent-offspring trios, revealed enrichment of rare damaging de novo mutations in chromatin modifier genes that play essential roles in brain and vascular development. Other VOGM probands harbored rare inherited damaging mutations in Ephrin signaling genes, including a genome-wide significant mutation burden in EPHB4. Inherited mutations showed incomplete penetrance and variable expressivity, with mutation carriers often exhibiting cutaneous vascular abnormalities, suggesting a two-hit mechanism. The identified mutations collectively account for ∼30% of studied VOGM cases. These findings provide insight into disease biology and may have clinical implications for risk assessment.
Abstract.
Zhang H, Zhang J, Zhang Q, Kim J, Zhang S, Gauthier SA, Spincemaille P, Nguyen TD, Sabuncu M, Wang Y, et al (2019). RSANet: Recurrent Slice-Wise Attention Network for Multiple Sclerosis Lesion Segmentation.
Abstract:
RSANet: Recurrent Slice-Wise Attention Network for Multiple Sclerosis Lesion Segmentation
Abstract.
Huang H, Song S, Banerjee S, Jiang T, Zhang J, Kahle KT, Sun D, Zhang Z (2019). The WNK-SPAK/OSR1 kinases and the cation-chloride cotransporters as therapeutic targets for neurological diseases. Aging and Disease, 10
2018
Bhuiyan MIH, Huang H, Zhang T, Molyneaux BJ, Poloyac SM, Zhang J, Deng X, Sun D, Sun D (2018). Abstract P198: WNK-SPAK-NKCC1 Cascade Activation Contributes to Worsened Brain Damage in Mice with Hypertension Co-Morbidity after Ischemic Stroke.
Hypertension,
72(Suppl_1).
Abstract:
Abstract P198: WNK-SPAK-NKCC1 Cascade Activation Contributes to Worsened Brain Damage in Mice with Hypertension Co-Morbidity after Ischemic Stroke
. Objectives:
. The WNK-SPAK/OSR1 kinase complex plays an important role in renal salt handling and pathogenesis of hypertension by regulating ion transporters and channels. Hypertension is the most common risk factor for stroke and stroke patients with hypertension comorbidity have worsened outcome with an increased risk of dependency or death. However, the mechanisms underlying the worsened ischemic stroke pathophysiology with hypertension comorbidity remain poorly defined. In this study, we investigated roles of the WNK-SPAK-NKCC1 signaling pathway in ischemic brain damage in mice with hypertension comorbidity.
.
.
. Methods:
. Hypertension was induced in C57BL/6j male mouse (12-15 weeks) by subcutaneous infusion of 1000 ng/kg/min angiotensin II (AngII, mini-osmotic pump) for two weeks. Permanent ischemic stroke was induced by permanent occlusion of the distal branches of the left middle cerebral artery (pd-MCAO). Brain tissues were harvested for immunoblot assessment of expression levels of NKCC1, SPAK/OSR1 or WNK1-4. Infarct volume and hemisphere swelling were determined by TTC staining, and behavioral deficits were analyzed by foot fault test, cylinder test and adhesive tape removal test.
.
.
. Results:
. pd-MCAO stimulated expression of WNK proteins (isoforms 1, 2, 4), total and phosphorylated SPAK/OSR1 and NKCC1 proteins in ischemic brains of the AngII-infused hypertensive mice compared to normotensive saline controls. In parallel with the increased activation of WNK-SPAK-NKCC1 signaling, hypertensive mice displayed significantly larger infarct volume and hemispheric swelling at 24 h after pd-MCAO compared to normotensive controls. Moreover, hypertensive mice exhibited a slow recovery of neurological function after ischemic stroke compared to normotensive counterparts as assessed by sensory-motor sensitive tests.
.
.
. Conclusions:
. These results suggest that activation of the WNK-SPAK-NKCC1 complex in hypertensive ischemic brains associates, at least in part, with the worsened brain damage and neurological deficits. Pharmacological inhibition of WNK-SPAK complex has therapeutic potentials for stroke therapy with hypertension comorbidity.
.
Abstract.
Pinkas DM, Bufton JC, Bartual SG, Chen Z, Daubner GM, Schumacher F-R, Georghiou G, Zhang J, Sorrell FJ, Kurz T, et al (2018). Human with No Lysine Kinase 3 (WNK3). A Target Enabling Package, Version 4, 1-15.
Zhang J, Bhuiyan MIH, Zhang T, Karimy J, Wu Z, Kahle KT, Sun D, Deng X (2018). Restoration of brain water homeostasis and neurological function via a novel kinase-cotransporter modulator. Cell Symposia, Aging and Metabolism. 23rd - 25th Sep 2018.
Abstract:
Restoration of brain water homeostasis and neurological function via a novel kinase-cotransporter modulator
Abstract.
Wang J, Erazo T, Ferguson FM, Buckley DL, Gomez N, Muñoz-Guardiola P, Diéguez-Martínez N, Deng X, Hao M, Massefski W, et al (2018). Structural and atropisomeric factors governing the selectivity of pyrimido-benzodiazipinones as inhibitors of kinases and bromodomains.
ACS Chemical BiologyAbstract:
Structural and atropisomeric factors governing the selectivity of pyrimido-benzodiazipinones as inhibitors of kinases and bromodomains
© 2018 American Chemical Society. Bromodomains have been pursued intensively over the past several years as emerging targets for the devel-opment of anti-cancer and anti-inflammatory agents. It has recently been shown that some kinase inhibitors are able to potently inhibit the bromodomains of BRD4. The clinical activities of PLK inhibitor BI-2536 and JAK2-FLT3 inhibitor TG101348 have been attributed to this unexpected poly-pharmacology, indicating that dual-kinase/bromodomain activity may be advantageous in a therapeutic context. However, for target validation and biological investigation, a more selec-tive target profile is desired. Here we report that benzo[e]pyrimido-[5,4-b]diazepine-6(11H)-ones, versatile ATP-site di-rected kinase pharmacophores utilized in the development of inhibitors of multiple kinases including a number of previ-ously reported kinase chemical probes, are also capable of exhibiting potent BRD4-dependent pharmacology. Using a dual kinase-bromodomain inhibitor of the kinase domains of ERK5 and LRRK2, and the bromodomain of BRD4 as a case study, we define the structure-activity relationships required to achieve dual kinase/BRD4 activity as well as how to di-rect selectivity towards inhibition of either ERK5 or BRD4. This effort resulted in identification of one of the first report-ed kinase-selective chemical probes for ERK5 (JWG-071), a BET selective inhibitor with 1 μM BRD4 IC50 (JWG-115), and additional inhibitors with rationally designed polypharmacology (JWG-047, JWG-069). Co-crystallography of seven representative inhibitors with the first bromodomain of BRD4 demonstrate that distinct atropisomeric conformers rec-ognize the kinase ATP-site and the BRD4 acetyl lysine binding site, conformational preferences supported by rigid dock-ing studies.
Abstract.
Dumon C, Diabira D, Chudotvorova I, Bader F, Sahin S, Zhang J, Porcher C, Wayman G, Medina I, Gaiarsa J-L, et al (2018). The adipocyte hormone leptin sets the emergence of hippocampal inhibition in mice. eLife, 7, e36726-e36726.
Bhuiyan MIH, Huang H, Zhang T, Molyneaux BJ, Poloyac SM, Zhang J, Deng X, Sun D (2018). WNK-SPAK-NKCC1 Cascade Activation Contributes to Worsened Brain Damage in Mice with Hypertension Comorbidity after Ischemic Stroke. Joint Hypertension 2018 Scientific Sessions. 6th - 9th Sep 2018.
Abstract:
WNK-SPAK-NKCC1 Cascade Activation Contributes to Worsened Brain Damage in Mice with Hypertension Comorbidity after Ischemic Stroke
Abstract.
2017
Karimy JK, Zhang J, Kurland DB, Theriault BC, Duran D, Furey CG, Gerzanich V, Simard JM, Kahle KT (2017). 166 TLR-4-Regulated Cerebrospinal Fluid Hypersecretion in Post-Hemorrhagic Hydrocephalus. Neurosurgery, 64(CN_suppl_1), 242-242.
Zhang J, Burgess JG (2017). Enhanced eicosapentaenoic acid production by a new deep-sea marine bacterium Shewanella electrodiphila MAR441(T).
PLOS ONE,
12(11).
Author URL.
Heubl M, Zhang J, Pressey JC, Al Awabdh S, Renner M, Gomez-Castro F, Moutkine I, Eugène E, Russeau M, Kahle KT, et al (2017). GABAA receptor dependent synaptic inhibition rapidly tunes KCC2 activity via the Cl--sensitive WNK1 kinase.
Nat Commun,
8(1).
Abstract:
GABAA receptor dependent synaptic inhibition rapidly tunes KCC2 activity via the Cl--sensitive WNK1 kinase.
The K+-Cl- co-transporter KCC2 (SLC12A5) tunes the efficacy of GABAA receptor-mediated transmission by regulating the intraneuronal chloride concentration [Cl-]i. KCC2 undergoes activity-dependent regulation in both physiological and pathological conditions. The regulation of KCC2 by synaptic excitation is well documented; however, whether the transporter is regulated by synaptic inhibition is unknown. Here we report a mechanism of KCC2 regulation by GABAA receptor (GABAAR)-mediated transmission in mature hippocampal neurons. Enhancing GABAAR-mediated inhibition confines KCC2 to the plasma membrane, while antagonizing inhibition reduces KCC2 surface expression by increasing the lateral diffusion and endocytosis of the transporter. This mechanism utilizes Cl- as an intracellular secondary messenger and is dependent on phosphorylation of KCC2 at threonines 906 and 1007 by the Cl--sensing kinase WNK1. We propose this mechanism contributes to the homeostasis of synaptic inhibition by rapidly adjusting neuronal [Cl-]i to GABAAR activity.
Abstract.
Author URL.
Karimy JK, Zhang J, Kurland DB, Theriault BC, Duran D, Stokum JA, Furey CG, Zhou X, Mansuri MS, Montejo J, et al (2017). Inflammation-dependent cerebrospinal fluid hypersecretion by the choroid plexus epithelium in posthemorrhagic hydrocephalus.
Nature Medicine,
23(8), 997-1003.
Abstract:
Inflammation-dependent cerebrospinal fluid hypersecretion by the choroid plexus epithelium in posthemorrhagic hydrocephalus
© 2017 Nature America, Inc. part of Springer Nature. All rights reserved. The choroid plexus epithelium (CPE) secretes higher volumes of fluid (cerebrospinal fluid, CSF) than any other epithelium and simultaneously functions as the blood-CSF barrier to gate immune cell entry into the central nervous system. Posthemorrhagic hydrocephalus (PHH), an expansion of the cerebral ventricles due to CSF accumulation following intraventricular hemorrhage (IVH), is a common disease usually treated by suboptimal CSF shunting techniques. PHH is classically attributed to primary impairments in CSF reabsorption, but little experimental evidence supports this concept. In contrast, the potential contribution of CSF secretion to PHH has received little attention. In a rat model of PHH, we demonstrate that IVH causes a Toll-like receptor 4 (TLR4)-and NF-κ B-dependent inflammatory response in the CPE that is associated with a 3-fold increase in bumetanide-sensitive CSF secretion. IVH-induced hypersecretion of CSF is mediated by TLR4-dependent activation of the Ste20-type stress kinase SPAK, which binds, phosphorylates, and stimulates the NKCC1 co-transporter at the CPE apical membrane. Genetic depletion of TLR4 or SPAK normalizes hyperactive CSF secretion rates and reduces PHH symptoms, as does treatment with drugs that antagonize TLR4-NF-κ B signaling or the SPAK-NKCC1 co-transporter complex. These data uncover a previously unrecognized contribution of CSF hypersecretion to the pathogenesis of PHH, demonstrate a new role for TLRs in regulation of the internal brain milieu, and identify a kinase-regulated mechanism of CSF secretion that could be targeted by repurposed US Food and Drug Administration (FDA)-approved drugs to treat hydrocephalus.
Abstract.
Karimy J, Zhang J, Medzhitov R, Simard M, Kahle K (2017). Inflammation-dependent cerebrospinal fluid hypersecretion from the choroid plexus in post-hemorrhagic hydrocephalus. 42nd FEBS congress. 10th - 14th Sep 2017.
Abstract:
Inflammation-dependent cerebrospinal fluid hypersecretion from the choroid plexus in post-hemorrhagic hydrocephalus
Abstract.
Zhang J, Karimy JK, Delpire E, Kahle KT (2017). Pharmacological targeting of SPAK kinase in disorders of impaired epithelial transport.
Expert Opinion on Therapeutic Targets,
21(8), 795-804.
Abstract:
Pharmacological targeting of SPAK kinase in disorders of impaired epithelial transport
© 2017 Informa UK Limited, trading as Taylor. &. Francis Group. Introduction: the mammalian SPS1-related proline/alanine-rich serine-threonine kinase SPAK (STK39) modulates ion transport across and between epithelial cells in response to environmental stimuli such osmotic stress and inflammation. Research over the last decade has established a central role for SPAK in the regulation of ion and water transport in the distal nephron, colonic crypts, and pancreatic ducts, and has implicated deregulated SPAK signaling in NaCl-sensitive hypertension, ulcerative colitis and Crohn’s disease, and cystic fibrosis. Areas covered: We review recent advances in our understanding of the role of SPAK kinase in the regulation of epithelial transport. We highlight how SPAK signaling–including its upstream Cl–sensitive activators, the WNK kinases, and its downstream ion transport targets, the cation- Cl–cotransporters contribute to human disease. We discuss prospects for the pharmacotherapeutic targeting of SPAK kinase in specific human disorders that feature impaired epithelial homeostasis. Expert opinion: the development of novel drugs that antagonize the SPAK-WNK interaction, inhibit SPAK kinase activity, or disrupt SPAK kinase activation by interfering with its binding to MO25α/β could be useful adjuncts in essential hypertension, inflammatory colitis, and cystic fibrosis.
Abstract.
Shekarabi M, Zhang J, Khanna AR, Ellison DH, Delpire E, Kahle KT (2017). WNK Kinase Signaling in Ion Homeostasis and Human Disease. Cell Metabolism, 25(2), 285-299.
2016
Kahle K, Schmouth J-F, Lavastre V, Latremoliere A, Zhang J, Andrews N, Dion PA, Duan J, Woolf CJ, Inquimbert P, et al (2016). Antagonism of Wnk/Hsn2 kinase ameliorates neuropathic pain by reducing maladaptive KCC2 inhibitory phosphorylation after nerve injury. 84th AANS Annual Scientific Meeting. 30th Apr - 4th May 2016.
Abstract:
Antagonism of Wnk/Hsn2 kinase ameliorates neuropathic pain by reducing maladaptive KCC2 inhibitory phosphorylation after nerve injury
Abstract.
Kahle K, Schmouth J-F, Lavastre V, Latremoliere A, Zhang J, Andrews N, Dion PA, Duan J, Woolf CJ, Inquimbert P, et al (2016). Antagonism of Wnk/Hsn2 kinase ameliorates neuropathic pain by reducing maladaptive KCC2 inhibitory phosphorylation after nerve injury.
Author URL.
Andrukhova O, Zhang J, Alessi D, Erben R (2016). Bone loss in KLHL3 knock-in mice characterized by a pseudohypoaldosteronism type II-like phenotype is mediated by renal PTH resistance. Bone Abstracts
Zhang J, Gao G, Begum G, Wang J, Khanna AR, Shmukler BE, Daubner GM, de los Heros P, Davies P, Varghese J, et al (2016). Functional kinomics establishes a critical node of volume-sensitive cation-Cl− cotransporter regulation in the mammalian brain.
Scientific Reports,
6(1).
Abstract:
Functional kinomics establishes a critical node of volume-sensitive cation-Cl− cotransporter regulation in the mammalian brain
AbstractCell volume homeostasis requires the dynamically regulated transport of ions across the plasmalemma. While the ensemble of ion transport proteins involved in cell volume regulation is well established, the molecular coordinators of their activities remain poorly characterized. We utilized a functional kinomics approach including a kinome-wide siRNA-phosphoproteomic screen, a high-content kinase inhibitor screen, and a kinase trapping-Orbitrap mass spectroscopy screen to systematically identify essential kinase regulators of KCC3 Thr991/Thr1048 phosphorylation – a key signaling event in cell swelling-induced regulatory volume decrease (RVD). In the mammalian brain, we found the Cl−-sensitive WNK3-SPAK kinase complex, required for cell shrinkage-induced regulatory volume decrease (RVI) via the stimulatory phosphorylation of NKCC1 (Thr203/Thr207/Thr212), is also essential for the inhibitory phosphorylation of KCC3 (Thr991/Thr1048). This is mediated in vivo by an interaction between the CCT domain in SPAK and RFXV/I domains in WNK3 and NKCC1/KCC3. Accordingly, genetic or pharmacologic WNK3-SPAK inhibition prevents cell swelling in response to osmotic stress and ameliorates post-ischemic brain swelling through a simultaneous inhibition of NKCC1-mediated Cl− uptake and stimulation of KCC3-mediated Cl− extrusion. We conclude that WNK3-SPAK is an integral component of the long-sought “Cl−/volume-sensitive kinase” of the cation-Cl− cotransporters, and functions as a molecular rheostat of cell volume in the mammalian brain.
Abstract.
Kahle KT, Schmouth J-F, Lavastre V, Latremoliere A, Zhang J, Andrews N, Omura T, Laganière J, Rochefort D, Hince P, et al (2016). Inhibition of the kinase WNK1/HSN2 ameliorates neuropathic pain by restoring GABA inhibition.
Science Signaling,
9(421).
Abstract:
Inhibition of the kinase WNK1/HSN2 ameliorates neuropathic pain by restoring GABA inhibition
Mice lacking the HSN2 form of the kinase WNK1 are protected from neuropathic pain due to nerve injury.
Abstract.
Zhang J, Deng X, Kahle KT (2016). Leveraging unique structural characteristics of WNK kinases to achieve therapeutic inhibition.
Science Signaling,
9(450).
Abstract:
Leveraging unique structural characteristics of WNK kinases to achieve therapeutic inhibition
Exploiting unique characteristics of the WNK-SPAK pathway may yield useful antihypertensive medications.
Abstract.
Emami K, Nelson A, Hack E, Zhang J, Green DH, Caldwell GS, Mesbahi E (2016). MALDI-TOF Mass Spectrometry Discriminates Known Species and Marine Environmental Isolates of Pseudoalteromonas. Frontiers in Microbiology, 7
Kahle KT, Flores B, Bharucha-Goebel D, Zhang J, Donkervoort S, Hegde M, Begum G, Duran D, Liang B, Sun D, et al (2016). Peripheral motor neuropathy is associated with defective kinase regulation of the KCC3 cotransporter.
Science Signaling,
9(439).
Abstract:
Peripheral motor neuropathy is associated with defective kinase regulation of the KCC3 cotransporter
Evaluation of a patient with peripheral motor weakness reveals a key role for phosphorylation-dependent regulation of the transporter KCC3 in the peripheral nervous system.
Abstract.
2015
Kazlauskaite A, Martínez‐Torres RJ, Wilkie S, Kumar A, Peltier J, Gonzalez A, Johnson C, Zhang J, Hope AG, Peggie M, et al (2015). Binding to serine 65‐phosphorylated ubiquitin primes Parkin for optimal. <scp>PINK</scp>. 1‐dependent phosphorylation and activation. EMBO reports, 16(8), 939-954.
Schumacher F, Siew K, Zhang J, Johnson C, Wood N, Cleary SE, Al Maskari RS, Ferryman JT, Hardege I, Yasmin, et al (2015). Characterisation of the Cullin‐3 mutation that causes a severe form of familial hypertension and hyperkalaemia. EMBO Molecular Medicine, 7(10), 1285-1306.
Zhang J, Siew K, Macartney T, O'Shaughnessy KM, Alessi DR (2015). Critical role of the SPAK protein kinase CCT domain in controlling blood pressure.
Hum Mol Genet,
24(16), 4545-4558.
Abstract:
Critical role of the SPAK protein kinase CCT domain in controlling blood pressure.
The STE20/SPS1-related proline/alanine-rich kinase (SPAK) controls blood pressure (BP) by phosphorylating and stimulating the Na-Cl (NCC) and Na-K-2Cl (NKCC2) co-transporters, which regulate salt reabsorption in the kidney. SPAK possesses a conserved carboxy-terminal (CCT) domain, which recognises RFXV/I motifs present in its upstream activator [isoforms of the With-No-lysine (K) kinases (WNKs)] as well as its substrates (NCC and NKCC2). To define the physiological importance of the CCT domain, we generated knock-in mice in which the critical CCT domain Leu502 residue required for high affinity recognition of the RFXI/V motif was mutated to Alanine. The SPAK CCT domain defective knock-in animals are viable, and the Leu502Ala mutation abolished co-immunoprecipitation of SPAK with WNK1, NCC and NKCC2. The CCT domain defective animals displayed markedly reduced SPAK activity and phosphorylation of NCC and NKCC2 co-transporters at the residues phosphorylated by SPAK. This was also accompanied by a reduction in the expression of NCC and NKCC2 protein without changes in mRNA levels. The SPAK CCT domain knock-in mice showed typical features of Gitelman Syndrome with mild hypokalaemia, hypomagnesaemia, hypocalciuria and displayed salt wasting on switching to a low-Na diet. These observations establish that the CCT domain plays a crucial role in controlling SPAK activity and BP. Our results indicate that CCT domain inhibitors would be effective at reducing BP by lowering phosphorylation as well as expression of NCC and NKCC2.
Abstract.
Author URL.
Hatcher JM, Zhang J, Choi HG, Ito G, Alessi DR, Gray NS (2015). Discovery of a Pyrrolopyrimidine (JH-II-127), a Highly Potent, Selective, and Brain Penetrant LRRK2 Inhibitor.
ACS Med Chem Lett,
6(5), 584-589.
Abstract:
Discovery of a Pyrrolopyrimidine (JH-II-127), a Highly Potent, Selective, and Brain Penetrant LRRK2 Inhibitor.
Activating mutations in leucine-rich repeat kinase 2 (LRRK2) are present in a subset of Parkinson's disease (PD) patients and may represent an attractive therapeutic target. Here we report a 2-anilino-4-methylamino-5-chloropyrrolopyrimidine, JH-II-127 (18), as a potent and selective inhibitor of both wild-type and G2019S mutant LRRK2. Compound 18 substantially inhibits Ser910 and Ser935 phosphorylation of both wild-type LRRK2 and G2019S mutant at a concentration of 0.1-0.3 μM in a variety of cell types and is capable of inhibiting Ser935 phosphorylation in mouse brain following oral delivery of doses as low as 30 mg/kg.
Abstract.
Author URL.
Siew K, Zhang J, Schumacher F, Alessi DR, Kurtz T, O'Shaughnessy KM (2015). Pulse waveform analysis reveals vascular contributions to Gordon Syndrome and Gitelman Syndrome blood pressure homeostasis. Proceedings Abstracts of the Physiological Society. 1st - 1st Jul 2015.
Abstract:
Pulse waveform analysis reveals vascular contributions to Gordon Syndrome and Gitelman Syndrome blood pressure homeostasis
Abstract.
Zhang J, Burgess JG (2015). Shewanella electrodiphila sp. nov. a psychrotolerant bacterium isolated from Mid-Atlantic Ridge deep-sea sediments.
Int J Syst Evol Microbiol,
65(9), 2882-2889.
Abstract:
Shewanella electrodiphila sp. nov. a psychrotolerant bacterium isolated from Mid-Atlantic Ridge deep-sea sediments.
Strains MAR441(T) and MAR445 were isolated from Mid-Atlantic Ridge sediments from a depth of 2734 m, and were found to belong to the genus Shewanella. The strains were rod-shaped, pigmented, non-motile and capable of anaerobic growth either by fermentation of carbohydrates or by anaerobic respiration. The strains utilized a variety of electron acceptors, including nitrate and ferric compounds, and could utilize peptone when grown anaerobically in a two-chambered microbial fuel cell, which used carbon cloth electrodes and delivered a stable power output of ,150-200 mW m(-2). The major fatty acids were typical of the genus Shewanella, with major components C13 : 0, iso-C13 : 0, iso-C15 : 0, C16 : 0, C16 : 1ω7c, C18 : 1ω7c and C20 : 5ω3 fatty acids. The DNA G+C content of strains MAR441(T) and MAR445 was 42.4 mol%. 16S rRNA gene sequence analysis indicated that strains MAR441(T) and MAR445 were most closely related to Shewanella olleyana (sequence similarities 97.9% to the type strain). DNA-DNA hybridization demonstrated only 15.6-37.2% relatedness between strain MAR441(T) and the type strains of related species of the genus Shewanella. Phenotypic characteristics confirmed that these isolates constituted a novel species of the genus Shewanella, for which the name Shewanella electrodiphila sp. nov. is proposed; the type strain is MAR441(T) (5ATCC BAA-2408(T) = DSM 24955(T)).
Abstract.
Author URL.
Friedel P, Kahle KT, Zhang J, Hertz N, Pisella LI, Buhler E, Schaller F, Duan J, Khanna AR, Bishop PN, et al (2015). WNK1-regulated inhibitory phosphorylation of the KCC2 cotransporter maintains the depolarizing action of GABA in immature neurons.
Science Signaling,
8(383).
Abstract:
WNK1-regulated inhibitory phosphorylation of the KCC2 cotransporter maintains the depolarizing action of GABA in immature neurons
. Immature neurons need WNK1-dependent phosphorylation of KCC2 to prevent a premature switch in the Cl
. −
. gradient and the effect of GABA.
.
Abstract.
2014
Keith S, Jinwei Z, Dario R. A, Kevin M. O (2014). Disruption of STE20/SPS1-related Proline/Alanine-rich Kinase (SPAK) binding lowers blood pressure and recapitulates Gitelman syndrome in mice. American Society of Nephrology Kidney Week 2014 Annual Meeting.
Abstract:
Disruption of STE20/SPS1-related Proline/Alanine-rich Kinase (SPAK) binding lowers blood pressure and recapitulates Gitelman syndrome in mice
Abstract.
Kahle KT, Gen G, Zhang J, Latremoliere A, Andrews N, Shang Y, Alessi D, Woolf C, Elledge S (2014). Promoting Endogenous GABAergic Analgesia via Kinase Modulation of Neuronal Ion Plasticity. 2014 CNS ANNUAL MEETING. 1st - 1st Aug 2014.
Abstract:
Promoting Endogenous GABAergic Analgesia via Kinase Modulation of Neuronal Ion Plasticity
Abstract.
Kahle KT, Gao G, Zhang J, Latremoliere A, Andrews N, Shang Y, Alessi D, Woolf C, Elledge S, Clapham D, et al (2014). Ronald R. Tasker Young Investigator Award 165 Promoting Endogenous GABAergic Analgesia via Kinase Modulation of Neuronal Ion Plasticity. Neurosurgery, 61(Supplement 1), 214-214.
Heros PDL, Zhang J, Gourlay R, Campbell D, Deak M, Macartney T, Kahle K, Alessi D (2014). SPAK/OSR1 kinases directly phosphorylate the K+-Cl- co-transporters. Federation of American Societies for Experimental Biology. 1st - 1st Apr 2014.
Abstract:
SPAK/OSR1 kinases directly phosphorylate the K+-Cl- co-transporters
Abstract.
Heros P, Zhang J, Gourlay R, Campbell D, Deak M, Macartney T, Kahle K, Alessi D (2014). SPAK/OSR1 kinases directly phosphorylate the K+‐Cl‐ co‐transporters (1109.7). The FASEB Journal, 28(S1).
Alessi DR, Zhang J, Khanna A, Hochdörfer T, Shang Y, Kahle KT (2014). The WNK-SPAK/OSR1 pathway: Master regulator of cation-chloride cotransporters.
Science Signaling(334).
Abstract:
The WNK-SPAK/OSR1 pathway: Master regulator of cation-chloride cotransporters
The WNK-SPAK/OSR1 kinase complex is composed of the kinases WNK (with no lysine) and SPAK (SPS1-related proline/alanine-rich kinase) or the SPAK homolog OSR1 (oxidative stress-responsive kinase 1). The WNK family senses changes in intracellular Cl-concentration, extracellular osmolarity, and cell volume and transduces this information to sodium (Na+), potassium (K+), and chloride (Cl-) cotransporters [collectively referred to as CCCs (cation-chloride cotransporters)] and ion channels to maintain cellular and organismal homeostasis and affect cellular morphology and behavior. Several genes encoding proteins in this pathway aremutated in human disease, and the cotransporters are targets of commonly used drugs. WNKs stimulate the kinases SPAK and OSR1, which directly phosphorylate and stimulate Cl--importing, Na+-driven CCCs or inhibit the Cl--extruding, K+-driven CCCs. These coordinated and reciprocal actions on the CCCs are triggered by an interaction between RFXV/I motifs within the WNKs and CCCs and a conserved carboxyl-terminal docking domain in SPAK and OSR1. This interaction site represents a potentially druggable node that could bemore effective than targeting the cotransporters directly. In the kidney, WNK-SPAK/OSR1 inhibition decreases epithelial NaCl reabsorption and K+secretion to lower blood pressure while maintaining serum K+. In neurons, WNK-SPAK/OSR1 inhibition could facilitate Cl-extrusion and promote γ-aminobutyric acidergic (GABAergic) inhibition. Such drugs could have efficacy as K+-sparing blood pressure-lowering agents in essential hypertension, nonaddictive analgesics in neuropathic pain, and promoters of GABAergic inhibition in diseases associated with neuronal hyperactivity, such as epilepsy, spasticity, neuropathic pain, schizophrenia, and autism.
Abstract.
de Los Heros P, Alessi DR, Gourlay R, Campbell DG, Deak M, Macartney TJ, Kahle KT, Zhang J (2014). The WNK-regulated SPAK/OSR1 kinases directly phosphorylate and inhibit the K+ -Cl- co-transporters.
Biochemical Journal,
458(3), 559-573.
Abstract:
The WNK-regulated SPAK/OSR1 kinases directly phosphorylate and inhibit the K+ -Cl- co-transporters
Precise homoeostasis of the intracellular concentration of Cl- is achieved via the co-ordinated activities of the Cl- influx and efflux. We demonstrate that the WNK (WNK lysine-deficient protein kinase)-activated SPAK (SPS1-related proline/alanine-rich kinase)/OSR1 (oxidative stress-responsive kinase 1) knownto directly phosphorylate and stimulate the N[K]CCs (Na+-K+ ion co-transporters), also promote inhibition of the KCCs (K+-Cl- co-transporters) by directly phosphorylating a recently described C-terminal threonine residue conserved in allKCCisoforms [Site-2 (Thr1048)]. First, we demonstrate that SPAK and OSR1, in the presence of theMO25 regulatory subunit, robustly phosphorylates allKCC isoforms at Site-2 in vitro. Secondly, STOCK1S-50699, a WNK pathway inhibitor, suppresses SPAK/OSR1 activation and KCC3ASite-2 phosphorylationwith similar efficiency. Thirdly, in ES (embryonic stem) cells lacking SPAK/OSR1 activity, endogenous phosphorylation of KCC isoforms at Site-2 is abolished and these cells display elevated basal activity of 86Rb+ uptake that was not markedly stimulated further by hypotonic high K+ conditions, consistent with KCC3A activation. Fourthly, a tight correlation exists between SPAK/OSR1 activity and the magnitude of KCC3A Site-2 phosphorylation. Lastly, a Site-2 alanine KCC3A mutant preventing SPAK/OSR1 phosphorylation exhibits increased activity. We also observe that KCCs are directly phosphorylated by SPAK/OSR1, at a novel Site-3 (Thr5 in KCC1/KCC3 and Thr6 in KCC2/KCC4), and a previously recognized KCC3-specific residue, Site-4 (Ser96 ). These data demonstrate that the WNK-regulated SPAK/OSR1 kinases directly phosphorylate the N[K]CCs and KCCs, promoting their stimulation and inhibition respectively. Given these reciprocal actions with anticipated net effects of increasing Cl- influx, we propose that the targeting of WNK-SPAK/OSR1 with kinase inhibitors might be a novel potent strategy to enhance cellular Cl - extrusion, with potential implications for the therapeutic modulation of epithelial and neuronal ion transport in human disease states. © 2014 the Author(s).
Abstract.
2013
Delbroek L, Van Kolen K, Steegmans L, da Cunha R, Mandemakers W, Daneels G, De Bock PJ, Zhang J, Gevaert K, De Strooper B, et al (2013). Development of an enzyme-linked immunosorbent assay for detection of cellular and in vivo LRRK2 S935 phosphorylation.
Journal of Pharmaceutical and Biomedical Analysis,
76, 49-58.
Abstract:
Development of an enzyme-linked immunosorbent assay for detection of cellular and in vivo LRRK2 S935 phosphorylation
After the discovery of kinase activating mutations in leucine-rich repeat kinase 2 (LRRK2) as associated with autosomal dominant forms of Parkinson's disease, inhibition of the kinase is being extensively explored as a disease modifying strategy. As signaling properties and substrate(s) of LRRK2 are poorly documented, autophosphorylation has been an important readout for the enzyme's activity. Western blotting using anti-phospho-S910 or S935 LRRK2 antibodies showed effectiveness in demonstrating inhibitory effects of compounds. In this communication we describe two types of enzyme-linked immunosorbent assays (ELISA) to determine LRRK2 protein levels and kinase activity. Both assays take advantage of the sensitivity of the earlier described total and pS935 antibodies for detection (Nichols et al. Biochem. J. 2010) [10]. The first assay is based on anti-GFP-based capturing of overexpressed LRRK2 and is highly suitable to show cellular effects of kinase inhibitors in a 96-well format. In the other platform anti-LRRK2-based capturing allows detection of endogenously expressed LRRK2 in rat tissue with no significant signal in tissue from LRRK2 knockout rats. Furthermore, both assays showed a significant reduction in pS935 levels on cellular and transgenic R1441C/G LRRK2. With the anti-LRRK2 ELISA we were able to detect LRRK2 phosphorylation in human peripheral blood mononuclear cells (PBMC). To conclude, we report two sensitive assays to monitor LRRK2 expression and kinase activity in samples coming from cellular and in vivo experimental settings. Both can show their value in drug screening and biomarker development but will also be useful in the elucidation of LRRK2-mediated signaling pathways. © 2012 Elsevier B.V.
Abstract.
Yao C, Johnson WM, Gao Y, Wang W, Zhang J, Deak M, Alessi. DR, Zhu X, Mieyal JJ, Roder H, et al (2013). Kinase inhibitors arrest neurodegeneration in cell and C. elegans models of LRRK2 toxicity.
Human Molecular Genetics,
22(2), 328-344.
Abstract:
Kinase inhibitors arrest neurodegeneration in cell and C. elegans models of LRRK2 toxicity
Mutations in leucine-rich repeat kinase 2 (LRRK2) are the most frequent known cause of late-onset Parkinson's disease (PD). To explore the therapeutic potential of small molecules targeting the LRRK2 kinase domain, we characterized two LRRK2 kinase inhibitors, TTT-3002 and LRRK2-IN1, for their effects against LRRK2 activity in vitro and in Caenorhabditis elegans models of LRRK2-linked neurodegeneration. TTT-3002 and LRRK2-IN1 potently inhibited in vitro kinase activity of LRRK2 wild-type and mutant proteins, attenuated phosphorylation of cellular LRRK2 and rescued neurotoxicity of mutant LRRK2 in transfected cells. To establish whether LRRK2 kinase inhibitors can mitigate pathogenesis caused by different mutations including G2019S and R1441C located within and outside of the LRRK2 kinase domain, respectively, we evaluated effects of TTT-3002 and LRRK2-IN1 against R1441C- and G2019S-induced neurodegeneration in C. elegans models. TTT-3002 and LRRK2-IN1 rescued the behavioral deficit characteristic of dopaminergic impairment in transgenic C. elegans expressing human R1441C- and G2019S-LRRK2. The inhibitors displayed nanomolar to low micromolar rescue potency when administered either pre-symptomatically or post-symptomatically, indicating both prevention and reversal of the dopaminergic deficit. The same treatments also led to long-lasting prevention and rescue of neurodegeneration. In contrast, TTT-3002 and LRRK2-IN1 were ineffective against the neurodegenerative phenotype in transgenic worms carrying the inhibitor-resistant A2016T mutation ofLRRK2, suggesting that they elicit neuroprotective effects in vivo by targeting LRRK2 specifically. Our findings indicate that the LRRK2 kinase activity is critical for neurodegeneration caused by R1441C and G2019S mutations, suggesting that kinase inhibition of LRRK2 may represent a promising therapeutic strategy for PD. © the Author 2012. Published by Oxford University Press. All rights reserved.
Abstract.
Deng X, Elkins JM, Zhang J, Yang Q, Erazo T, Gomez N, Choi HG, Wang J, Dzamko N, Lee JD, et al (2013). Structural determinants for ERK5 (MAPK7) and leucine rich repeat kinase 2 activities of benzo[e]pyrimido-[5,4-b]diazepine-6(11H)-ones.
European Journal of Medicinal Chemistry,
70, 758-767.
Abstract:
Structural determinants for ERK5 (MAPK7) and leucine rich repeat kinase 2 activities of benzo[e]pyrimido-[5,4-b]diazepine-6(11H)-ones
The benzo[e]pyrimido-[5,4-b]diazepine-6(11H)-one core was discovered as a novel ERK5 (also known as MAPK7 and BMK1) inhibitor scaffold, previously. Further structure-activity relationship studies of this scaffold led to the discovery of ERK5-IN-1 (26) as the most selective and potent ERK5 inhibitor reported to date. 26 potently inhibits ERK5 biochemically with an IC 50 of 0.162 ± 0.006 μM and in cells with a cellular EC 50 for inhibiting epidermal growth factor induced ERK5 autophosphorylation of 0.09 ± 0.03 μM. Furthermore, 26 displays excellent selectivity over other kinases with a KINOMEscan selectivity score (S10) of 0.007, and exhibits exceptional bioavailability (F%) of 90% in mice. 26 will serve as a valuable tool compound to investigate the ERK5 signaling pathway and as a starting point for developing an ERK5 directed therapeutic agent. © 2013 Elsevier Masson SAS. All rights reserved.
Abstract.
2012
Choi HS, Zhang J, Deng X, Hatcher JM, Patricelli MP, Zhao Z, Alessi DR, Gray NS (2012). Brain penetrant LRRK2 inhibitor.
ACS Medicinal Chemistry Letters,
3(8), 658-662.
Abstract:
Brain penetrant LRRK2 inhibitor
Activating mutations in leucine-rich repeat kinase 2 (LRRK2) are present in a subset of Parkinson's disease (PD) patients and may represent an attractive therapeutic target. Here, we report that a 2-anilino-4-methylamino-5- chloropyrimidine, HG-10-102-01 (4), is a potent and selective inhibitor of wild-type LRRK2 and the G2019S mutant. Compound 4 substantially inhibits Ser910 and Ser935 phosphorylation of both wild-type LRRK2 and G2019S mutant at a concentration of 0.1-0.3 μM in cells and is the first compound reported to be capable of inhibiting Ser910 and Ser935 phosphorylation in mouse brain following intraperitoneal delivery of doses as low as 50 mg/kg. © 2012 American Chemical Society.
Abstract.
Zhang J, Deng X, Choi HG, Alessi DR, Gray NS (2012). Characterization of TAE684 as a potent LRRK2 kinase inhibitor.
Bioorganic and Medicinal Chemistry Letters,
22(5), 1864-1869.
Abstract:
Characterization of TAE684 as a potent LRRK2 kinase inhibitor
Leucine-rich repeat kinase 2 (LRRK2) is linked to Parkinson's disease and may represent an attractive therapeutic target. Here we report a 2,4-dianilino-5-chloro-pyrimidine, TAE684, a previously reported inhibitor of anaplastic lymphoma kinase (ALK), is also a potent inhibitor of LRRK2 kinase activity (IC50of 7.8 nM against wild-type LRRK2, 6.1 nM against the G2019S mutant). TAE684 substantially inhibits Ser910 and Ser935 phosphorylation of both wild-type LRRK2 and G2019S mutant at a concentration of 0.1-0.3 μM in cells and in mouse spleen and kidney, but not in brain, following oral doses of 10 mg/kg. © 2012 Elsevier Ltd. All rights reserved.
Abstract.
Zhang. J, Zhang E, Scott K, Burgess JG (2012). Enhanced electricity production by use of reconstituted artificial consortia of estuarine bacteria grown as biofilms.
Environmental Science and Technology,
46(5), 2984-2992.
Abstract:
Enhanced electricity production by use of reconstituted artificial consortia of estuarine bacteria grown as biofilms
Microbial fuel cells (MFCs) can convert organic compounds directly into electricity by catalytic oxidation, and although MFCs have attracted considerable interest, there is little information on the electricity-generating potential of artificial bacterial biofilms. We have used acetate-fed MFCs inoculated with sediment, with two-chamber bottles and carbon cloth electrodes to deliver a maximum power output of ∼175 mW•m -2 and a stable power output of ∼105 mW•m -2. Power production was by direct transfer of electrons to the anode from bacterial consortia growing on the anode, as confirmed by cyclic voltammetry (CV) and scanning electron microscopy (SEM). Twenty different species (74 strains) of bacteria were isolated from the consortium under anaerobic conditions and cultured in the laboratory, of which 34% were found to be exoelectrogens in single-species studies. Exoelectrogenesis by members of the genera Vibrio, Enterobacter, and Citrobacter and by Bacillus stratosphericus was confirmed, by use of culture-based methods, for the first time. An MFC with a natural bacterial consortium showed higher power densities than those obtained with single strains. In addition, the maximum power output could be further increased to ∼200 mW•m -2 when an artificial consortium consisting of the best 25 exoelectrogenic isolates was used, demonstrating the potential for increased performance and underlying the importance of artificial biofilms for increasing power output. © 2012 American Chemical Society.
Abstract.
Reith AD, Bamborough P, Jandu K, Andreotti D, Mensah L, Dossang P, Choi HG, Deng X, Zhang J, Alessi DR, et al (2012). GSK2578215A; a potent and highly selective 2-arylmethyloxy-5-substitutent- N-arylbenzamide LRRK2 kinase inhibitor.
Bioorganic and Medicinal Chemistry Letters,
22(17), 5625-5629.
Abstract:
GSK2578215A; a potent and highly selective 2-arylmethyloxy-5-substitutent- N-arylbenzamide LRRK2 kinase inhibitor
Leucine-rich repeat kinase 2 (LRRK2) is a promising therapeutic target for some forms of Parkinson's disease. Here we report the discovery and characterization of 2-arylmethyloxy-5-subtitutent-N-arylbenzamides with potent LRRK2 activities exemplified by GSK2578215A which exhibits biochemical IC 50s of around 10 nM against both wild-type LRRK2 and the G2019S mutant. GSK2578215A exhibits exceptionally high selectivity for LRRK2 across the kinome, substantially inhibits Ser910 and Ser935 phosphorylation of both wild-type LRRK2 and G2019S mutant at a concentration of 0.3-1.0 μM in cells and in mouse spleen and kidney, but not in brain, following intraperitoneal injection of 100 mg/kg. © 2012 Elsevier Ltd. All rights reserved.
Abstract.
2011
Zhang J, Zeng R (2011). Molecular cloning and expression of an extracellular α-amylase gene from an Antarctic deep sea psychrotolerant Pseudomonas stutzeri strain 7193.
World Journal of Microbiology and Biotechnology,
27(4), 841-850.
Abstract:
Molecular cloning and expression of an extracellular α-amylase gene from an Antarctic deep sea psychrotolerant Pseudomonas stutzeri strain 7193
Psychrotolerant Pseudomonas stutzeri strain 7193 capable of producing an extracellular α-amylase was isolated from deep sea sediments of Prydz Bay, Antarctic. The 59678-Da protein (AmyP) was encoded by 1665-bp gene (amyP). The deduced amino acid sequence was identified with four regions, which are conserved in amylolytic enzymes and form a catalytic domain, and was predicted to be maltotetraose forming extracellular amylase by using the I-TASSER online server. Purification of AmyP amylases from both the recombinant of Escherichia coli Top 10 F′ and strain 7193 was conducted. Biochemical characterization revealed that the optimal amylase activity was observed at pH 9.0 and temperature 40°C. The enzymes were unstable at temperatures above 30°C, and only retain half of their highest activity after incubation at 60°C for 5 min. Thin-layer chromatography analysis of the products of the amylolytic reaction showed the presence of maltotetraose, maltotriose, maltose and glucose in the starch hydrolysate. © 2010 Springer Science+Business Media B.V.
Abstract.
2008
Zhang J, Zeng R (2008). Molecular cloning and expression of a cold-adapted lipase gene from an antarctic deep sea psychrotrophic bacterium Pseudomonas sp. 7323.
Marine Biotechnology,
10(5), 612-621.
Abstract:
Molecular cloning and expression of a cold-adapted lipase gene from an antarctic deep sea psychrotrophic bacterium Pseudomonas sp. 7323
A psychrotrophic bacterium producing a cold-adapted lipase was isolated from the deep-sea sediment of Prydz Bay, Antarctic and identified as a Pseudomonas strain. Determination of the nucleotide sequence of the gene encoding a lipase from Pseudomonas sp. 7323 (lipA) revealed that LipA is composed of 617 amino acid residues with a calculated molecular weight of 64,466 Da. LipA has a GXSXG motif, which is conserved in lipases/esterases and generally contains the active-site serine. The lipase purified from the Escherichia coli transformant (rLipA) by metal-chelating chromatography exhibited the same electrophoretic mobility as did the wild-type lipase (wLipA) purified from strain 7323, and both enzymes were quite similar in physicochemical properties. The optimal temperature and pH value for the lipases activity were 30°C and 9.0, respectively. They were unstable at temperatures above 25°C and only retained half of their highest activity after incubation at 60°C for 5 min. These results indicated that the enzymes were typical alkaline cold-adapted enzymes. Both enzymes were particularly activated by Ca2+. Additionally, the enzymes hydrolyzed p-nitrophenyl caprate and tributyrin at the highest velocity among the other p-nitrophenyl esters and triglycerides. © 2008 Springer Science+Business Media, LLC.
Abstract.
Zhang J, Zeng R (2008). Purification and characterization of a cold-adapted α-amylase produced by Nocardiopsis sp. 7326 isolated from Prydz Bay, Antarctic.
Marine Biotechnology,
10(1), 75-82.
Abstract:
Purification and characterization of a cold-adapted α-amylase produced by Nocardiopsis sp. 7326 isolated from Prydz Bay, Antarctic
An actinomycete strain 7326 producing cold-adapted α-amylase was isolated from the deep sea sediment of Prydz Bay, Antarctic. It was identified as Nocardiopsis based on morphology, 16S rRNA gene sequence analysis, and physiological and biochemical characteristics. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and zymogram activity staining of purified amylase showed a single band equal to a molecular mass of about 55 kDa. The optimal activity temperature of Nocardiopsis sp. 7326 amylase was 35°C, and the activity decreased dramatically at temperatures above 45°C. The enzyme was stable between pH 5 and 10, and exhibited a maximal activity at pH 8.0. Ca2+, Mn2+, Mg2+, Cu2+, and Co2+stimulated the activity of the enzyme significantly, and Rb2+, Hg2+, and EDTA inhibited the activity. The hydrolysates of soluble starch by the enzyme were mainly glucose, maltose, and maltotriose. This is the first report on the isolation and characterization of cold-adapted amylase from Nocardiopsis sp. © 2007 Springer Science+Business Media, LLC.
Abstract.
2007
Zhang J, Lin S, Zeng R (2007). Cloning, expression, and characterization of a cold-adapted lipase gene from an antarctic deep-sea psychorotrophic bacterium, Psychobacter sp. 7195.
Journal of Microbiology and Biotechnology,
17(4), 604-610.
Abstract:
Cloning, expression, and characterization of a cold-adapted lipase gene from an antarctic deep-sea psychorotrophic bacterium, Psychobacter sp. 7195
A psychrotrophic strain 7195 showing extracellular lipolytic activity towards tributyrin was isolated from deep-sea sediment of Prydz Bay and identified as a Psychrobacter species. By screening a genomic DNA library of Psychrobacter sp. 7195, an open reading frame of 954 by coding for a lipase gene, lipA1, was identified, cloned, and sequenced. The deduced LipA1 consisted of 317 amino acids with a molecular mass of 35,210 kDa. It had one consensus motif, G-N-S-M-G (GXSXG), containing the putative active-site serine, which was conserved in other cold-adapted lipolytic enzymes. The recombinant LipA1 was purified by column chromatography with DEAE Sepharose CL-4B, and Sephadex G-75, and preparative polyacrylamide gel electrophoresis, in sequence. The purified enzyme showed highest activity at 30°C, and was unstable at temperatures higher than 30°C, indicating that it was a typical cold-adapted enzyme. The optimal pH for activity was 9.0, and the enzyme was stable between pH 7.0-10.0 after 24 h incubation at 4°C. The addition of Ca2+and Mg2+enhanced the enzyme activity of LipA1, whereas the Cd2+, Zn2+, Co2+, Fe3+, Hg2+, Fe2+, Rb2+, and EDTA strongly inhibited the activity. The LipA1 was activated by various detergents, such as Triton X-100, Tween 80, Tween 40, Span 60, Span 40, CHAPS, and SDS, and showed better resistance towards them. Substrate specificity analysis showed that there was a preference for trimyristin and p-nitrophenyl myristate (C14acyl groups). © the Korean Society for Microbiology and Biotechnology.
Abstract.
Zhang J, Zeng R (2007). Isolation of antarctic psychrotrophilic Pseudomonas sp. 7197 and cloning of ppa gene for inorganic pyrophosphatase (PPase).
Gaojishu Tongxin/Chinese High Technology Letters,
17(10), 1067-1071.
Abstract:
Isolation of antarctic psychrotrophilic Pseudomonas sp. 7197 and cloning of ppa gene for inorganic pyrophosphatase (PPase)
A psychrotrophile strain 7197, which produces inorganic pyrophosphatase (PPase), was isolated from the deep sea sediment of Prydz Bay, Antarctic. The morphology identification and 16S rDNA sequence analysis showed that the strain belongs to genus Pseudomonas. The ppa gene for inorganic pyrophosphatase (PPase) was cloned by PCR according to the primers. Nucleotide sequence analysis revealed an open reading frame (ORF) of 531 bp encoding the PPase. The amino acid sequence deduced from the nucleotide sequence of the ppa corresponded to a protein of 176 amino acid residues with a molecular weight of 19531 kDa. The PPase showed the highest amino acid sequence identity to the PPase of Psy-chrobacter sp. 273-4 with 97%, and had 79% and 75% of identity to the PPase of Neisseria meningitidis Z2491 and Mannheimia succiniciproducens MBEL55E respectively.
Abstract.
Zhang J, Zeng R (2007). Psychrotrophic amylolytic bacteria from deep sea sediment of Prydz Bay, Antarctic: Diversity and characterization of amylases.
World Journal of Microbiology and Biotechnology,
23(11), 1551-1557.
Abstract:
Psychrotrophic amylolytic bacteria from deep sea sediment of Prydz Bay, Antarctic: Diversity and characterization of amylases
Seventeen psychrotrophic bacteria with cold-adaptive amylolytic, lipolytic or proteolytic activity were isolated from deep sea sediment of Prydz Bay, Antarctic. They were affiliated with γ-Proteobacteria (12 strains) and gram-positive bacteria (5 strains) as determined by 16S rDNA sequencing. The amylase-producing strains belonged to genus Pseudomonas, Rhodococcus, and Nocardiopsis. Two Pseudomonas strains, 7193 and 7197, which showed highest amylolytic activity were chosen for further study. The optimal temperatures for their growth and amylase-producing were between 15 and 20°C. Both of the purified amylases showed highest activity at 40°C and pH 9.0, and retained 50% activity at 5°C. The SDS-PAGE and zymogram activity staining showed that the molecular mass of strain 7193 and 7197 amylases were about 60 and 50 kDa respectively. The Pseudomonas sp. 7193 amylase hydrolyzed soluble starch into glucose, maltose, maltotriose, and maltotetraose, indicating that it had both activities of α-amylase and glucoamylase. The product hydrolyzed by Pseudomonas sp. 7197 amylase was meltotetraose. © 2007 Springer Science+Business Media B.V.
Abstract.
2006
Zhang J, Zeng R (2006). Cloning, expression and characterization of the cold active lipase (Lip3) from metagenomic DNA of an antarctic deep sea sediment.
Progress in Biochemistry and Biophysics,
33(12), 1207-1214.
Abstract:
Cloning, expression and characterization of the cold active lipase (Lip3) from metagenomic DNA of an antarctic deep sea sediment
The metagenomic DNA was extracted from the deep sea sediment with the depth of 900m of Prydz Bay, Antarctic. A lipase gene (lip3) with the size of 948bp was cloned from the metagenomic DNA by PCR with the primers designed. The deduced Lip3 protein was composed of 315 amino acids (AA) with a molecular mass of 34.577 ku. The motifs GFGNS(GXGXS)and G-N-S-M-G(GXSXG)in the AA sequences of Lip3 were found to be conserved in other lipase. They were most conserved sequence among the serine hydrolase and were necessary for the activity. A 35 ku of Lip3 was purified by Ni-NTA chelating sepharose column from the extract of recombinant E.coli Top 10F′ cell harboring a pLLP-OmpA plasmid inserted with lip3. The purified Lip3 was most active at 25°C and kept 22% of activity at 0°C. Only 10% of activity was retained after it was incubated at 35°C for 60 min. The optimal pH value for the Lip3 activity was 8.0. The Kmvalue of the enzyme towards p-nitrophenyl palmitate increased with the increasing of assayed temperature. These results indicated that Lip3 was a typical alkaline cold active enzyme.
Abstract.
Zhang J, Lian M, Zeng R (2006). Screening, fermentation condition and enzyme characterization of multicomponent enzymes producing Pseudomonas sp. NJ197.
Chinese Journal of Applied and Environmental Biology,
12(5), 683-687.
Abstract:
Screening, fermentation condition and enzyme characterization of multicomponent enzymes producing Pseudomonas sp. NJ197
A strain NJ197, producing cold-adapted complex enzyme, was isolated from deep sea sediment of the Prydz Bay, Antarctic. The morphological identification and 16S rDNA sequence analysis showed that it belonged to genus Pseudomonas. The optimal growth temperature of the Pseudomonas sp. NJ197 was 5-15°C, indicating that it was a psychro-tolerant bacterium. The strain could use various single carbonaceous and nitrogenous substances to produce complex enzyme. Its optimal and highest temperatures for enzyme production were 20°C and 30°C, respectively. The enzyme purification was performed by ammonium sulfate fractionation and anion exchange chromatography with DEAE cellulose-52. The activities of lipase and amylase were the strongest among the complex enzymes produced by Pseudomonas sp. NJ197, and the both were sensitive to high temperature. The optimal temperature and pH value for the lipase activity were 30°C and 9.0, while those for the amylase activity were 35°C and 9.5, respectively. The results indicated that they were typical alkaline cold-adapted enzymes, whose activities were stimulated by Ca2+, Mn2+, Cu2+, Co2+and Fe3+, and inhibited by Zn2+, Hg2+, Rb2+, Cd2+and EDTA. The cold-adapted lipase showed better resistance to inactivation of 1% detergents, such as SDS and CHAPS.
Abstract.
