跨膜蛋白16A(transmembrane protein 16A,TMEM16A)是一种钙激活的氯离子通道(calcium-activated chloride channel,CaCC),它在肿瘤中广泛过表达,对肿瘤的发生发展有着重要的影响。在各种肿瘤中,TMEM16A的过表达不仅可以促进癌细胞的增...跨膜蛋白16A(transmembrane protein 16A,TMEM16A)是一种钙激活的氯离子通道(calcium-activated chloride channel,CaCC),它在肿瘤中广泛过表达,对肿瘤的发生发展有着重要的影响。在各种肿瘤中,TMEM16A的过表达不仅可以促进癌细胞的增殖、转移和侵袭,抑制肿瘤细胞的凋亡,影响肿瘤的生长,还与肿瘤的治疗耐药、复发和预后不良有关。因此,探索TMEM16A的体内表达情况与肿瘤各个方面的关系具有重要的意义,该综述旨在总结近十年来TMEM16A在肿瘤中的研究进展,为以后TMEM16A在肿瘤中的进一步研究提供新的思路。展开更多
Anoctamin 1(ANO1) or TMEM16 A gene encodes a member of Ca^(2+) activated Cl^(-) channels(CaCCs) that are critical for physiological functions,such as epithelial secretion,smooth muscle contraction and sensory signal t...Anoctamin 1(ANO1) or TMEM16 A gene encodes a member of Ca^(2+) activated Cl^(-) channels(CaCCs) that are critical for physiological functions,such as epithelial secretion,smooth muscle contraction and sensory signal transduction.The attraction and interest in ANO1/TMEM16 A arise from a decade long investigations that abnormal expression or dysfunction of ANO1 is involved in many pathological phenotypes and diseases,including asthma,neuropathic pain,hypertension and cancer.However,the lack of specific modulators of ANO1 has impeded the efforts to validate ANO1 as a therapeutic target.This review focuses on the recent progress made in understanding of the pathophysiological functions of CaCC ANO1 and the current modulators used as pharmacological tools,hopefully illustrating a broad spectrum of ANO1 channelopathy and a path forward for this target validation.展开更多
Vascular smooth muscle cell(VSMC) migration plays a critical role in the pathogenesis of many cardiovascular diseases.We recently showed that TMEM16 A is involved in hypertensioninduced cerebrovascular remodeling.Howe...Vascular smooth muscle cell(VSMC) migration plays a critical role in the pathogenesis of many cardiovascular diseases.We recently showed that TMEM16 A is involved in hypertensioninduced cerebrovascular remodeling.However,it is unclear whether this effect is related to the regulation of VSMC migration.Here,we investigated whether and how TMEM16 A contributes to migration in basilar artery smooth muscle cells(BASMCs).We observed that AngⅡ increased the migration of cultured BASMCs,which was markedly inhibited by overexpression of TMEM16 A.TMEM16 A overexpression inhibited AngⅡ-induced RhoA/ROCK2 activation,and myosin light chain phosphatase(MLCP)and myosin light chain(MLC20) phosphorylation.But AnsⅡ-induced myosin light chain kinase(MLCK)activation was not affected by TMEM16 A.Furthermore,a suppressed activation of integrinβ3/FAK pathway,determined by reduced integrinβ3 expression,FAK phosphorylation and F-actin rearrangement,was observed in TMEM16 A-overexpressing BASMCs upon AngⅡ stimulation.Contrary to the results of TMEM16 A overexpression,silencing of TMEM16 A showed the opposite effects.These in vitro results were further demonstrated in vivo in basilar arteries from VSMC-specific TMEM10 A transgenic mice during AngⅡ-induced hypertension.Moreover,we observed that the inhibitory effect of TMEM16 A on BASMC migration was mediated by decreasing the activation of WNK1,a Cl^(-)-sensitive serine/threonine kinase.In conclusion,this study demonstrated that TMEM16 A suppressed AnsⅡ-induced BASMC migration,thus contributing to the protection against cerebrovascular remodeling during AngⅡ-infused hypertension.TMEM16 A may exert this effect by suppressing the RhoA/ROCK2/MLCP/MLC20 and integrinβ3/FAK signaling pathways via inhibiting WNK1.Our results suggest that TMEM16 A may serve as a novel therapeutic target for VSMC migration-related diseases,such as vascular remodeling.展开更多
Uncontrolled cell migration is a common feature of tumor metastasis and formation. Understanding the molecular targets critically involved in cell migration process can lead to the development of potentially novel the...Uncontrolled cell migration is a common feature of tumor metastasis and formation. Understanding the molecular targets critically involved in cell migration process can lead to the development of potentially novel therapeutic strategies for controlling invasion of tumor cells. In this study, we showed that calcium-activated chloride channel ANO 1/TMEM16A played an important role in cell migration and inhibition of ANO1 channel function suppressed the migration of tumor epithelial cells. Silencing ANO 1 by small hairpin RNA （shRNA） resulted in suppression of cell migration and invasiveness in cancer cell lines. In addition, pharmacological inhibition of ANO1 by the channel specific inhibitor T16Ain-A01 significantly slowed down the migration and invasion of tumor epithelial cells in a dose-dependent manner. Taken together, our findings have demonstrated that calcium-activated chloride channel ANO1 contributes to cell migration, and specific ANO1/TMEM16A inhibitors can be the promising candidate to develop new therapies for cancer metastasis.展开更多
TMEM16F is involved in many physiological processes such as blood coagulation,cell membrane fusion and bone mineralization.Activation of TMEM16F has been studied in various central nervous system diseases.High TMEM16F...TMEM16F is involved in many physiological processes such as blood coagulation,cell membrane fusion and bone mineralization.Activation of TMEM16F has been studied in various central nervous system diseases.High TMEM16F level has been also found to participate in microglial phagocytosis and transformation.Microglia-mediated neuroinflammation is a key factor in promoting the progression of Alzheimer’s disease.However,few studies have examined the effects of TMEM16F on neuroinflammation in Alzheimer’s disease.In this study,we established TMEM16F-knockdown AD model in vitro and in vivo to investigate the underlying regulatory mechanism about TMEM16F-mediated neuroinflammation in AD.We performed a Morris water maze test to evaluate the spatial memory ability of animals and detected markers for the microglia M1/M2 phenotype and NLRP3 inflammasome.Our results showed that TMEM16F was elevated in 9-month-old APP/PS1 mice.After TMEM16F knockdown in mice,spatial memory ability was improved,microglia polarization to the M2 phenotype was promoted,NLRP3 inflammasome activation was inhibited,cell apoptosis and Aβplaque deposition in brain tissue were reduced,and brain injury was alleviated.We used amyloid-beta(Aβ_(25-35))to stimulate human microglia to construct microglia models of Alzheimer’s disease.The levels of TMEM16F,inducible nitric oxide synthase(iNOS),proinflammatory cytokines and NLRP3 inflammasome-associated biomarkers were higher in Aβ_(25-35) treated group compared with that in the control group.TMEM16F knockdown enhanced the expression of the M2 phenotype biomarkers Arg1 and Socs3,reduced the release of proinflammatory factors interleukin-1,interleukin-6 and tumor necrosis factor-α,and inhibited NLRP3 inflammasome activation through reducing downstream proinflammatory factors interleukin-1βand interleukin-18.This inhibitory effect of TMEM16F knockdown on M1 microglia was partially reversed by the NLRP3 agonist Nigericin.Our findings suggest that TMEM16F participates in neuroinflammation in Alzheimer’s disease through participating in polarization of microglia and activation of the NLRP3 inflammasome.These results indicate that TMEM16F inhibition may be a potential therapeutic approach for Alzheimer’s disease treatment.展开更多
Anoctamin 1(ANO1)is a kind of calcium-activated chloride channel involved in nerve depolarization.ANO1 inhibitors display significant analgesic activity by the local peripheral and intrathecal administration.In this s...Anoctamin 1(ANO1)is a kind of calcium-activated chloride channel involved in nerve depolarization.ANO1 inhibitors display significant analgesic activity by the local peripheral and intrathecal administration.In this study,several thiophenecarboxylic acid and benzoic acid derivatives were identified as novel ANO1 inhibitors through the shape-based virtual screening,among which the 4-arylthiophene-3-carboxylic acid analogues with the best ANO1 inhibitory activity were designed,synthesized and compound 42(IC;=0.79μmol/L)was finally obtained.Compound 42 selectively inhibited ANO1 without affecting ANO2 and intracellular Ca;concentration.Subsequently,the analgesic effect was investigated by intragastric administration in pain models.Compound 42 significantly attenuated allodynia which was induced by formalin and chronic constriction injury.Through homology modeling and molecular dynamics,the binding site was predicted to be located near the calcium-binding region betweenα6 andα8.Our study validates ANO1 inhibitors having a significant analgesic effect by intragastric administration and also provides selective molecular tools for ANO1-related research.展开更多
文摘跨膜蛋白16A(transmembrane protein 16A,TMEM16A)是一种钙激活的氯离子通道(calcium-activated chloride channel,CaCC),它在肿瘤中广泛过表达,对肿瘤的发生发展有着重要的影响。在各种肿瘤中,TMEM16A的过表达不仅可以促进癌细胞的增殖、转移和侵袭,抑制肿瘤细胞的凋亡,影响肿瘤的生长,还与肿瘤的治疗耐药、复发和预后不良有关。因此,探索TMEM16A的体内表达情况与肿瘤各个方面的关系具有重要的意义,该综述旨在总结近十年来TMEM16A在肿瘤中的研究进展,为以后TMEM16A在肿瘤中的进一步研究提供新的思路。
基金supported by grants from the National Natural Science foundation of China (81573410)the Ministry of Science and Technology of China (2018ZX09711001-004-006)+2 种基金Shandong Provincial Medicine and Health Technology Development Plan (2019WS145, China)Medical Research Guidance Plan of Qingdao Municipal Health Committee (2019-WJZD091, China)Qingdao Excellent Young Medical Talent Training Project。
文摘Anoctamin 1(ANO1) or TMEM16 A gene encodes a member of Ca^(2+) activated Cl^(-) channels(CaCCs) that are critical for physiological functions,such as epithelial secretion,smooth muscle contraction and sensory signal transduction.The attraction and interest in ANO1/TMEM16 A arise from a decade long investigations that abnormal expression or dysfunction of ANO1 is involved in many pathological phenotypes and diseases,including asthma,neuropathic pain,hypertension and cancer.However,the lack of specific modulators of ANO1 has impeded the efforts to validate ANO1 as a therapeutic target.This review focuses on the recent progress made in understanding of the pathophysiological functions of CaCC ANO1 and the current modulators used as pharmacological tools,hopefully illustrating a broad spectrum of ANO1 channelopathy and a path forward for this target validation.
基金supported by the National Natural Science Foundation of China (grant Nos. 81872858, 81773721, 81573422, and 81903598)。
文摘Vascular smooth muscle cell(VSMC) migration plays a critical role in the pathogenesis of many cardiovascular diseases.We recently showed that TMEM16 A is involved in hypertensioninduced cerebrovascular remodeling.However,it is unclear whether this effect is related to the regulation of VSMC migration.Here,we investigated whether and how TMEM16 A contributes to migration in basilar artery smooth muscle cells(BASMCs).We observed that AngⅡ increased the migration of cultured BASMCs,which was markedly inhibited by overexpression of TMEM16 A.TMEM16 A overexpression inhibited AngⅡ-induced RhoA/ROCK2 activation,and myosin light chain phosphatase(MLCP)and myosin light chain(MLC20) phosphorylation.But AnsⅡ-induced myosin light chain kinase(MLCK)activation was not affected by TMEM16 A.Furthermore,a suppressed activation of integrinβ3/FAK pathway,determined by reduced integrinβ3 expression,FAK phosphorylation and F-actin rearrangement,was observed in TMEM16 A-overexpressing BASMCs upon AngⅡ stimulation.Contrary to the results of TMEM16 A overexpression,silencing of TMEM16 A showed the opposite effects.These in vitro results were further demonstrated in vivo in basilar arteries from VSMC-specific TMEM10 A transgenic mice during AngⅡ-induced hypertension.Moreover,we observed that the inhibitory effect of TMEM16 A on BASMC migration was mediated by decreasing the activation of WNK1,a Cl^(-)-sensitive serine/threonine kinase.In conclusion,this study demonstrated that TMEM16 A suppressed AnsⅡ-induced BASMC migration,thus contributing to the protection against cerebrovascular remodeling during AngⅡ-infused hypertension.TMEM16 A may exert this effect by suppressing the RhoA/ROCK2/MLCP/MLC20 and integrinβ3/FAK signaling pathways via inhibiting WNK1.Our results suggest that TMEM16 A may serve as a novel therapeutic target for VSMC migration-related diseases,such as vascular remodeling.
基金The Ministry of Science and Technology of China(Grant No.2014ZX09507003-006-004)
文摘Uncontrolled cell migration is a common feature of tumor metastasis and formation. Understanding the molecular targets critically involved in cell migration process can lead to the development of potentially novel therapeutic strategies for controlling invasion of tumor cells. In this study, we showed that calcium-activated chloride channel ANO 1/TMEM16A played an important role in cell migration and inhibition of ANO1 channel function suppressed the migration of tumor epithelial cells. Silencing ANO 1 by small hairpin RNA （shRNA） resulted in suppression of cell migration and invasiveness in cancer cell lines. In addition, pharmacological inhibition of ANO1 by the channel specific inhibitor T16Ain-A01 significantly slowed down the migration and invasion of tumor epithelial cells in a dose-dependent manner. Taken together, our findings have demonstrated that calcium-activated chloride channel ANO1 contributes to cell migration, and specific ANO1/TMEM16A inhibitors can be the promising candidate to develop new therapies for cancer metastasis.
基金supported by the National Natural Science Foundation of China,No.82072941(to QHX)Liaoning Province Key R&D Program Guidance Project,No.2020JH2/10300044Science and Technology Plan Project of Shenyang,No.20-205-4-050(both to XHS)。
文摘TMEM16F is involved in many physiological processes such as blood coagulation,cell membrane fusion and bone mineralization.Activation of TMEM16F has been studied in various central nervous system diseases.High TMEM16F level has been also found to participate in microglial phagocytosis and transformation.Microglia-mediated neuroinflammation is a key factor in promoting the progression of Alzheimer’s disease.However,few studies have examined the effects of TMEM16F on neuroinflammation in Alzheimer’s disease.In this study,we established TMEM16F-knockdown AD model in vitro and in vivo to investigate the underlying regulatory mechanism about TMEM16F-mediated neuroinflammation in AD.We performed a Morris water maze test to evaluate the spatial memory ability of animals and detected markers for the microglia M1/M2 phenotype and NLRP3 inflammasome.Our results showed that TMEM16F was elevated in 9-month-old APP/PS1 mice.After TMEM16F knockdown in mice,spatial memory ability was improved,microglia polarization to the M2 phenotype was promoted,NLRP3 inflammasome activation was inhibited,cell apoptosis and Aβplaque deposition in brain tissue were reduced,and brain injury was alleviated.We used amyloid-beta(Aβ_(25-35))to stimulate human microglia to construct microglia models of Alzheimer’s disease.The levels of TMEM16F,inducible nitric oxide synthase(iNOS),proinflammatory cytokines and NLRP3 inflammasome-associated biomarkers were higher in Aβ_(25-35) treated group compared with that in the control group.TMEM16F knockdown enhanced the expression of the M2 phenotype biomarkers Arg1 and Socs3,reduced the release of proinflammatory factors interleukin-1,interleukin-6 and tumor necrosis factor-α,and inhibited NLRP3 inflammasome activation through reducing downstream proinflammatory factors interleukin-1βand interleukin-18.This inhibitory effect of TMEM16F knockdown on M1 microglia was partially reversed by the NLRP3 agonist Nigericin.Our findings suggest that TMEM16F participates in neuroinflammation in Alzheimer’s disease through participating in polarization of microglia and activation of the NLRP3 inflammasome.These results indicate that TMEM16F inhibition may be a potential therapeutic approach for Alzheimer’s disease treatment.
基金supported by the National Key Research and Development Project(Grant No.2019YFC1708900)the National Natural Science Foundation of China(Grant No.21772005)+1 种基金National Major Scientific and Technological Special Project for Significant New Drugs Development(2019ZX09204-001,China)Beijing Municipal Natural Science Foundation(7202088,7172118,China)
文摘Anoctamin 1(ANO1)is a kind of calcium-activated chloride channel involved in nerve depolarization.ANO1 inhibitors display significant analgesic activity by the local peripheral and intrathecal administration.In this study,several thiophenecarboxylic acid and benzoic acid derivatives were identified as novel ANO1 inhibitors through the shape-based virtual screening,among which the 4-arylthiophene-3-carboxylic acid analogues with the best ANO1 inhibitory activity were designed,synthesized and compound 42(IC;=0.79μmol/L)was finally obtained.Compound 42 selectively inhibited ANO1 without affecting ANO2 and intracellular Ca;concentration.Subsequently,the analgesic effect was investigated by intragastric administration in pain models.Compound 42 significantly attenuated allodynia which was induced by formalin and chronic constriction injury.Through homology modeling and molecular dynamics,the binding site was predicted to be located near the calcium-binding region betweenα6 andα8.Our study validates ANO1 inhibitors having a significant analgesic effect by intragastric administration and also provides selective molecular tools for ANO1-related research.
