The prevalence of cardiovascular diseases(CVDs)has increased markedly as the world population has aged.Long non-coding RNAs(lncRNAs)have been reported as novel regulators in diverse pathophysiological conditions.Here,...The prevalence of cardiovascular diseases(CVDs)has increased markedly as the world population has aged.Long non-coding RNAs(lncRNAs)have been reported as novel regulators in diverse pathophysiological conditions.Here,we performed RNA sequencing(RNA-seq)and observed that the lncRNA Zeb1os1(zinc finger E-box binding homeobox 1,opposite strand 1),which is known as ZEB1-AS1(zinc finger E-box binding homeobox 1 antisense 1)in humans,was upregulated in the aged mice hearts,senescent cardiomyocytes,and human blood from elderly individuals.The human blood ZEB1-AS1 level was positively relevant to human age but negatively relevant to peak E to peak A(E/A).Silencing Zeb1os1 ameliorated diastolic dysfunction and cardiac senescence in aged mice.On the other hand,Zeb1os1 overexpression triggered cardiac dysfunction resembling that observed in aged mice.Mechanistically,we provide compelling evidence that Zeb1os1 interacts with the transient receptor potential mucolipin 1(TRPML1)for ubiquitination(UB)-mediated degradation.This process inhibits lysosomal Ca^(2+)efflux,impairing lysosome function.In addition,the functional domain of Zeb1os1,which contains the key nucleotides responsible for the pro-senescence property of full-length Zeb1os1 in cardiomyocytes.Together,these data suggest that Zeb1os1 is a potential target for ameliorating lysosomal dysfunction and aging-related cardiac impairment.展开更多
雷帕霉素(Rapamycin,Rap)是一种免疫抑制剂,在临床上主要应用于器官移植过程中的抗排异反应,同时,Rap在抗癌、神经保护和抗衰老等领域也展现出巨大潜力。通过抑制哺乳动物Rap靶蛋白(mammalian target of Rapamycin,mTOR)活性,Rap能够活...雷帕霉素(Rapamycin,Rap)是一种免疫抑制剂,在临床上主要应用于器官移植过程中的抗排异反应,同时,Rap在抗癌、神经保护和抗衰老等领域也展现出巨大潜力。通过抑制哺乳动物Rap靶蛋白(mammalian target of Rapamycin,mTOR)活性,Rap能够活化调节溶酶体功能的转录因子EB(transcription factor EB,TFEB)从而上调溶酶体功能,同时,Rap也能够去除mTOR对ULK1(unc-51 like autophagy activating kinase 1)的抑制效果从而促进自噬。最新研究显示,Rap能够直接激活溶酶体钙离子通道TRPML1,此效应并不依赖于mTOR;TRPML1通道的开放可释放溶酶体钙离子,钙调神经磷酸酶(Calcineurin)作为此溶酶体来源的钙信号的感受器活化TFEB,进而增强溶酶体功能并促进自噬。此发现拓宽了对Rap药理学机制的认知,为衰老等领域的机制研究提供了理论基础。本文将以经典的Rap-mTOR-ULK1/TFEB信号通路为导引,详细介绍溶酶体钙离子通道TRPML1被确认为Rap新靶标的论证过程,并简要讨论此新信号通路(Rap-TRPML1-Calcineurin-TFEB)的药理学意义。展开更多
Objective: ATP6V0d2 is a subunit of the vacuolar-type H+-ATPase (V-ATPase) that pumps H+ ions into lysosomes. TRPML1 (MCOLN1/Mcoln1) transports cations out of lysosomes.Mcoln1^(-/-) mice recapitulate the lysosomal sto...Objective: ATP6V0d2 is a subunit of the vacuolar-type H+-ATPase (V-ATPase) that pumps H+ ions into lysosomes. TRPML1 (MCOLN1/Mcoln1) transports cations out of lysosomes.Mcoln1^(-/-) mice recapitulate the lysosomal storage disorder mucolipidosis type IV (MLIV) phenotype. We previously demonstrated thatMcoln1^(-/-) female mice quickly became infertile at 5 months old (5M) with degenerating corpora lutea (CL) and progesterone (P4) deficiency. We tested our hypothesis thatAtp6v0d2 deficiency could partially compensate forMcoln1 deficiency to restore CL functions inAtp6v0d2^(-/-)Mcoln1^(-/-) mice.Methods: Control andAtp6v0d2^(-/-)Mcoln1^(-/-) female mice underwent fertility test from 2M to 7M. A subset of them was dissected at 5M on day 3.5 post-coitum (D3.5). The D3.5 ovaries from 5M control,Mcoln1^(-/-), andAtp6v0d2^(-/-)Mcoln1^(-/-) mice were evaluated for CL morphology, lipid droplet staining, and markers of mitochondria and P4 steroidogenesis in the luteal cells.Results: The fertility test ofAtp6v0d2^(-/-)Mcoln1^(-/-) female mice (2M–7M) revealed normal mating activity but reduced fertility compared with the control;yet ~25% of them remained fertile at 5M to 7M but with dystocia. We analyzed a subset of 11Atp6v0d2^(-/-)Mcoln1^(-/-) mice (5M) in the fertility test on D3.5: three (27.3%) had normal P4 levels and all examined CL parameters, indicating full restoration of CL function compared withMcoln1^(-/-), whereas eight had P4 deficiency, with two (18.2%) infertile and six (54.5%) once fertile. In contrast toMcoln1^(-/-) CLs, which had extensive amorphous cellular debris, indicating cell degeneration,Atp6v0d2^(-/-)Mcoln1^(-/-) CLs had reduced amorphous cellular debris regardless of P4 levels. However, similar toMcoln1^(-/-) CLs, P4-deficientAtp6v0d2^(-/-)Mcoln1^(-/-) CLs showed impaired differentiation, enlarged lipid droplets, disorganized expression of endothelial basal lamina marker collagen IV, and reduced expression of mitochondrial marker heat shock protein 60 (HSP60) and steroidogenesis rate-limiting protein StAR, indicating that additionalAtp6v0d2 deficiency compensates forMcoln1 deficiency-induced cell degeneration, but is insufficient to restore luteal cell differentiation and P4 steroidogenesis in P4-deficientAtp6v0d2^(-/-)Mcoln1^(-/-) CLs.Conclusion: This study shows thatAtp6v0d2^(-/-)Mcoln1^(-/-) CLs had varied improvements compared withMcoln1^(-/-) CLs, and it providesin vivo genetic evidence of the coordination between different lysosomal channels in CL function.展开更多
Objective:FemaleMcoln1^(-/-)mice exhibit progressive progesterone(P4)deficiency,luteal cell degeneration,and premature embryo implantation failure at 5 months old.We attempted to rescue embryo implantation in non-virg...Objective:FemaleMcoln1^(-/-)mice exhibit progressive progesterone(P4)deficiency,luteal cell degeneration,and premature embryo implantation failure at 5 months old.We attempted to rescue embryo implantation in non-virginMcoln1^(-/-)mice(5-6 months old)with exogenous P4 treatment on days 1.5 post-coitum(D1.5),D2.5,and D3.5,and observed partially restored luteal cell morphology on D4.5,but unexpectedly found 17β-estradiol(E2)contamination in the P4 working solution.In this study,we aim to investigate exogenous P4 and/or E2 for the partial recovery of luteal cell morphology in infertileMcoln1^(-/-)mice.Methods:Control and non-virginMcoln1^(-/-)mice(5-6 months old)were treated with newly ordered vehicle,P4,E2,or P4+E2 on D1.5 and D2.5 and dissected on D3.5 for P4 and E2 measurements,ovary histology,immunofluorescence,lipid droplet staining,and transmission electron microscopy.Results:E2 treatment significantly increased serum P4 levels in D3.5Mcoln1^(-/-)mice.E2 and P4+E2 treatments,but not P4 treatment alone,largely improved the morphology of D3.5Mcoln1^(-/-)corpora lutea,indicated by a more contiguous web-like collagen IV expression pattern,increased heat shock protein 60 expression,and reduced accumulation of large lipid droplets.Transmission electron microscopy revealed extremely enlarged autophagosomes and lipid droplets,lysosomes with lamellar structures,and mitochondria with reduced cristae in vehicle-treated D3.5Mcoln1^(-/-)luteal cells,while in E2-treated D3.5Mcoln1^(-/-)luteal cells,extremely enlarged autophagosomes and lipid droplets were reduced,indicating improved luteal cell ultrastructure.Conclusion:These findings reveal protective effects of high levels of exogenous E2 on P4 production and lysosomal function inMcoln1^(-/-)luteal cells.展开更多
基金funded by the National Natural Science Foundation of China(82273919,82270396,and U21A20339)the China Postdoctoral Science Foundation(2023T160176)。
文摘The prevalence of cardiovascular diseases(CVDs)has increased markedly as the world population has aged.Long non-coding RNAs(lncRNAs)have been reported as novel regulators in diverse pathophysiological conditions.Here,we performed RNA sequencing(RNA-seq)and observed that the lncRNA Zeb1os1(zinc finger E-box binding homeobox 1,opposite strand 1),which is known as ZEB1-AS1(zinc finger E-box binding homeobox 1 antisense 1)in humans,was upregulated in the aged mice hearts,senescent cardiomyocytes,and human blood from elderly individuals.The human blood ZEB1-AS1 level was positively relevant to human age but negatively relevant to peak E to peak A(E/A).Silencing Zeb1os1 ameliorated diastolic dysfunction and cardiac senescence in aged mice.On the other hand,Zeb1os1 overexpression triggered cardiac dysfunction resembling that observed in aged mice.Mechanistically,we provide compelling evidence that Zeb1os1 interacts with the transient receptor potential mucolipin 1(TRPML1)for ubiquitination(UB)-mediated degradation.This process inhibits lysosomal Ca^(2+)efflux,impairing lysosome function.In addition,the functional domain of Zeb1os1,which contains the key nucleotides responsible for the pro-senescence property of full-length Zeb1os1 in cardiomyocytes.Together,these data suggest that Zeb1os1 is a potential target for ameliorating lysosomal dysfunction and aging-related cardiac impairment.
基金funded by NIH R01HD065939(co-funded by ORWH and NICHD)NIH R03HD097384NIH R03 HD100652 to X.Y.
文摘Objective: ATP6V0d2 is a subunit of the vacuolar-type H+-ATPase (V-ATPase) that pumps H+ ions into lysosomes. TRPML1 (MCOLN1/Mcoln1) transports cations out of lysosomes.Mcoln1^(-/-) mice recapitulate the lysosomal storage disorder mucolipidosis type IV (MLIV) phenotype. We previously demonstrated thatMcoln1^(-/-) female mice quickly became infertile at 5 months old (5M) with degenerating corpora lutea (CL) and progesterone (P4) deficiency. We tested our hypothesis thatAtp6v0d2 deficiency could partially compensate forMcoln1 deficiency to restore CL functions inAtp6v0d2^(-/-)Mcoln1^(-/-) mice.Methods: Control andAtp6v0d2^(-/-)Mcoln1^(-/-) female mice underwent fertility test from 2M to 7M. A subset of them was dissected at 5M on day 3.5 post-coitum (D3.5). The D3.5 ovaries from 5M control,Mcoln1^(-/-), andAtp6v0d2^(-/-)Mcoln1^(-/-) mice were evaluated for CL morphology, lipid droplet staining, and markers of mitochondria and P4 steroidogenesis in the luteal cells.Results: The fertility test ofAtp6v0d2^(-/-)Mcoln1^(-/-) female mice (2M–7M) revealed normal mating activity but reduced fertility compared with the control;yet ~25% of them remained fertile at 5M to 7M but with dystocia. We analyzed a subset of 11Atp6v0d2^(-/-)Mcoln1^(-/-) mice (5M) in the fertility test on D3.5: three (27.3%) had normal P4 levels and all examined CL parameters, indicating full restoration of CL function compared withMcoln1^(-/-), whereas eight had P4 deficiency, with two (18.2%) infertile and six (54.5%) once fertile. In contrast toMcoln1^(-/-) CLs, which had extensive amorphous cellular debris, indicating cell degeneration,Atp6v0d2^(-/-)Mcoln1^(-/-) CLs had reduced amorphous cellular debris regardless of P4 levels. However, similar toMcoln1^(-/-) CLs, P4-deficientAtp6v0d2^(-/-)Mcoln1^(-/-) CLs showed impaired differentiation, enlarged lipid droplets, disorganized expression of endothelial basal lamina marker collagen IV, and reduced expression of mitochondrial marker heat shock protein 60 (HSP60) and steroidogenesis rate-limiting protein StAR, indicating that additionalAtp6v0d2 deficiency compensates forMcoln1 deficiency-induced cell degeneration, but is insufficient to restore luteal cell differentiation and P4 steroidogenesis in P4-deficientAtp6v0d2^(-/-)Mcoln1^(-/-) CLs.Conclusion: This study shows thatAtp6v0d2^(-/-)Mcoln1^(-/-) CLs had varied improvements compared withMcoln1^(-/-) CLs, and it providesin vivo genetic evidence of the coordination between different lysosomal channels in CL function.
基金the Offfce of the Vice President for Research,Interdisciplinary Toxicology Program,and Department of Physiology and Pharmacology at the University of Georgia,and the National Institutes of Health(grants NIH R01HD065939[co-funded by ORWH and NICHD],R03HD097384,and R03 HD100652 to X.Y.)for ffnancial supportSerum P4 and E2 levels were determined at The University of Virginia Center for Research in Reproduction Ligand Assay and Analysis Core,which is supported by the Eunice Kennedy Shriver NICHD/NIH(NCTRI)Grant P50-HD28934.
文摘Objective:FemaleMcoln1^(-/-)mice exhibit progressive progesterone(P4)deficiency,luteal cell degeneration,and premature embryo implantation failure at 5 months old.We attempted to rescue embryo implantation in non-virginMcoln1^(-/-)mice(5-6 months old)with exogenous P4 treatment on days 1.5 post-coitum(D1.5),D2.5,and D3.5,and observed partially restored luteal cell morphology on D4.5,but unexpectedly found 17β-estradiol(E2)contamination in the P4 working solution.In this study,we aim to investigate exogenous P4 and/or E2 for the partial recovery of luteal cell morphology in infertileMcoln1^(-/-)mice.Methods:Control and non-virginMcoln1^(-/-)mice(5-6 months old)were treated with newly ordered vehicle,P4,E2,or P4+E2 on D1.5 and D2.5 and dissected on D3.5 for P4 and E2 measurements,ovary histology,immunofluorescence,lipid droplet staining,and transmission electron microscopy.Results:E2 treatment significantly increased serum P4 levels in D3.5Mcoln1^(-/-)mice.E2 and P4+E2 treatments,but not P4 treatment alone,largely improved the morphology of D3.5Mcoln1^(-/-)corpora lutea,indicated by a more contiguous web-like collagen IV expression pattern,increased heat shock protein 60 expression,and reduced accumulation of large lipid droplets.Transmission electron microscopy revealed extremely enlarged autophagosomes and lipid droplets,lysosomes with lamellar structures,and mitochondria with reduced cristae in vehicle-treated D3.5Mcoln1^(-/-)luteal cells,while in E2-treated D3.5Mcoln1^(-/-)luteal cells,extremely enlarged autophagosomes and lipid droplets were reduced,indicating improved luteal cell ultrastructure.Conclusion:These findings reveal protective effects of high levels of exogenous E2 on P4 production and lysosomal function inMcoln1^(-/-)luteal cells.
