Both cholinergic dysfunction and protein citrullination are the hallmarks of rheumatoid arthritis(RA),but the relationship between the two phenomena remains unclear.We explored whether and how cholinergic dysfunction ...Both cholinergic dysfunction and protein citrullination are the hallmarks of rheumatoid arthritis(RA),but the relationship between the two phenomena remains unclear.We explored whether and how cholinergic dysfunction accelerates protein citrullination and consequently drives the development of RA.Cholinergic function and protein citrullination levels in patients with RA and collageninduced arthritis(CIA)mice were collected.In both neuron-macrophage coculture system and CIA mice,the effect of cholinergic dysfunction on protein citrullination and expression of peptidylarginine deiminases(PADs)was assessed by immunofluorescence.The key transcription factors for PAD4 expression were predicted and validated.Cholinergic dysfunction in the patients with RA and CIA mice negatively correlated with the degree of protein citrullination in synovial tissues.The cholinergic or alpha7 nicotinic acetylcholine receptor(a7nAChR)deactivation and activation resulted in the promotion and reduction of protein citrullination in vitro and in vivo,respectively.Especially,the activation deficiency of a7nAChR induced the earlier onset and aggravation of CIA.Furthermore,deactivation of a7nAChR increased the expression of PAD4 and specificity protein-3(SP3)in vitro and in vivo.Our results suggest that cholinergic dysfunction-induced deficient a7nAChR activation,which induces the expression of SP3 and its downstream molecule PAD4,accelerating protein citrullination and the development of RA.展开更多
Parkinson’s disease(PD)is one of the most common neurodegenerative disorders of aging,characterized by the degeneration of dopamine neurons(DA neurons)in the substantial nigra,leading to the advent of both motor symp...Parkinson’s disease(PD)is one of the most common neurodegenerative disorders of aging,characterized by the degeneration of dopamine neurons(DA neurons)in the substantial nigra,leading to the advent of both motor symptoms and non-motor symptoms.Current treatments include electrical stimulation of the affected brain areas and dopamine replacement therapy.Even though both categories are effective in treating PD patients,the disease progression cannot be stopped.The research advance into cell therapies provides exciting potential for the treatment of PD.Current cell sources include neural stem cells(NSCs)from fetal brain tissues,human embryonic stem cells(hESCs),induced pluripotent stem cells(iPSCs)and directly induced dopamine neurons(iDA neurons).Here,we evaluate the research progress in different cell sources with a focus on using iPSCs as a valuable source and propose key challenges for developing cells suitable for large-scale clinical applications in the treatment of PD.展开更多
The combination of disease-specific human induced pluripotent stem cells(iPSC)and directed cell differentiation offers an ideal platform for modeling and studying many inherited human diseases.Wilson’s disease(WD)is ...The combination of disease-specific human induced pluripotent stem cells(iPSC)and directed cell differentiation offers an ideal platform for modeling and studying many inherited human diseases.Wilson’s disease(WD)is a monogenic disorder of toxic copper accumulation caused by pathologic mutations of the ATP7B gene.WD affects multiple organs with primary manifestations in the liver and central nervous system(CNS).In order to better investigate the cellular pathogenesis of WD and to develop novel therapies against various WD syndromes,we sought to establish a comprehensive platform to differentiate WD patient iPSC into both hepatic and neural lineages.Here we report the generation of patient iPSC bearing a Caucasian population hotspot mutation of ATP7B.Combining with directed cell differentiation strategies,we successfully differentiated WD iPSC into hepatocyte-like cells,neural stem cells and neurons.Gene expression analysis and cDNA sequencing confirmed the expression of the mutant ATP7B gene in all differentiated cells.Hence we established a platform for studying both hepatic and neural abnormalities of WD,which may provide a new tool for tissue-specific disease modeling and drug screening in the future.展开更多
Oxidative damage to cells leads to accumulated harmful wastes,which in turn aggravate the imbalance of reactive oxygen species(ROS)and related diseases.Therefore,provoking the cellular defense system against severe ox...Oxidative damage to cells leads to accumulated harmful wastes,which in turn aggravate the imbalance of reactive oxygen species(ROS)and related diseases.Therefore,provoking the cellular defense system against severe oxidation and maintaining ROS homeostasis are desired.Herein,we designed and synthesized a powerful mitochondria-targeting aggregation-induced emission photosensitizer(named DTCSPY)by maximal restriction of heat dissipation.It is demonstrated that taming ROS generation within mitochondria through photosensitization-triggered autophagy via DTCSPY achieved a better neuroprotective effect against oxidative damages than Nacety-L-cysteine and vitamin C.This work not only provides a new way to design high-performance photosensitizers by regulating the photophysical property,but also verifies the concept that taming ROS can be used for cell protection against destructive oxidation,thereby displaying broad prospects for alleviating oxidation-related diseases and promoting cell-based therapy.展开更多
Neurodegeneration with brain iron accumulation is a broad term that describes a heterogeneous group of progressive and invalidating neurologic disorders in which iron deposits in certain brain areas,mainly the basal g...Neurodegeneration with brain iron accumulation is a broad term that describes a heterogeneous group of progressive and invalidating neurologic disorders in which iron deposits in certain brain areas,mainly the basal ganglia.The predominant clinical symptoms include spasticity,progressive dystonia,Parkinson's disease-like symptoms,neuropsychiatric alterations,and retinal degeneration.Among the neurodegeneration with brain iron accumulation disorders,the most frequent subtype is pantothenate kinase-associated neurodegeneration(PKAN) caused by defects in the gene encoding the enzyme pantothenate kinase 2(PANK2)which catalyzed the first reaction of the coenzyme A biosynthesis pathway.Currently there is no effective treatment to prevent the inexorable course of these disorders.The aim of this review is to open up a discussion on the utility of using cellular models derived from patients as a valuable tool for the development of precision medicine in PKAN.Recently,we have described that dermal fibroblasts obtained from PKAN patients can manifest the main pathological changes of the disease such as intracellular iron accumulation accompanied by large amounts of lipofuscin granules,mitochondrial dysfunction and a pronounced increase of markers of oxidative stress.In addition,PKAN fibroblasts showed a morphological senescence-like phenotype.Interestingly,pantothenate supplementation,the substrate of the PANK2 enzyme,corrected all pathophysiological alterations in responder PKAN fibroblasts with low/residual PANK2 enzyme expression.However,pantothenate treatment had no favourable effect on PKAN fibroblasts harbouring mutations associated with the expression of a truncated/incomplete protein.The correction of pathological alterations by pantothenate in individual mutations was also verified in induced neurons obtained by direct reprograming of PKAN fibroblasts.Our observations indicate that pantothenate supplementation can increase/stabilize the expression levels of PANK2 in specific mutations.Fibroblasts and induced neuron展开更多
Background: The ε4 allele of apolipoprotein E (APOE ε4) is the strongest known genetic risk factor for late-onset Alzheimer’s disease (AD), associated with amyloid pathogenesis. However, it is not clear how APOE ε...Background: The ε4 allele of apolipoprotein E (APOE ε4) is the strongest known genetic risk factor for late-onset Alzheimer’s disease (AD), associated with amyloid pathogenesis. However, it is not clear how APOE ε4 accelerates amyloid-beta (Aβ) deposition during the seeding stage of amyloid development in AD patient neurons. Methods: AD patient induced neurons (iNs) with an APOE ε4 inducible system were prepared from skin fibroblasts of AD patients. Transcriptome analysis was performed using RNA isolated from the AD patient iNs expressing APOE ε4 at amyloid-seeding and amyloid-aggregation stages. Knockdown of IGFBP3 was applied in the iNs to investigate the role of IGFBP3 in the APOE ε4-mediated amyloidosis. Results: We optimized amyloid seeding stage in the iNs of AD patients that transiently expressed APOE ε4. Remarka-bly, we demonstrated that Aβ pathology was aggravated by the induction of APOE ε4 gene expression at the amyloid early-seeding stage in the iNs of AD patients. Moreover, transcriptome analysis in the early-seeding stage revealed that IGFBP3 was functionally important in the molecular pathology of APOE ε4-associated AD. Conclusions: Our findings suggest that the presence of APOE ε4 at the early Aβ-seeding stage in patient iNs is critical for aggravation of sporadic AD pathology. These results provide insights into the importance of APOE ε4 expression for the progression and pathogenesis of sporadic AD.展开更多
In vitro studies have demonstrated that many factors of bone morphogenetic proteins （BMPs） induce cholinergic differentiation of neural stem cells. However, BMP retains the potential to induce increased numbers of c...In vitro studies have demonstrated that many factors of bone morphogenetic proteins （BMPs） induce cholinergic differentiation of neural stem cells. However, BMP retains the potential to induce increased numbers of cholinergic neurons in central nervous system regions that are rich in cholinergic cells, which is an important determinant of BMP. Therefore, BMP-4 was added to neural stem cell culture medium or the adult rat hippocampal dentate gyrus. Results demonstrated that BMP-4 induced cholinergic differentiation of neural stem cells in vitro and increased the number of cholinergic neurons in the adult rat hippocampal dentate gyrus.展开更多
Background:Investigations of the pathogenic mechanisms in motor neurons(MNs)derived from amyotrophic lateral sclerosis(ALS)disease-specific induced pluripotent stem(iPS)cell lines could improve understanding of the is...Background:Investigations of the pathogenic mechanisms in motor neurons(MNs)derived from amyotrophic lateral sclerosis(ALS)disease-specific induced pluripotent stem(iPS)cell lines could improve understanding of the issues affecting MNs.Therefore,in this study we explored mutant superoxide dismutase 1(SOD1)protein expression in MNs derived from the iPS cell lines of ALS patients carrying different SOD1 mutations.Methods:We generated induced pluripotent stem cell(iPSC)lines from two familial ALS(FALS)patients withSOD1-V14M andSOD1-C111Y mutations,and then differentiated them into MNs.We investigated levels of the SOD1 protein in iPSCs and MNs,the intracellular Ca2+levels in MNs,and the lactate dehydrogenase(LDH)activity in the process of differentiation into the MNs derived from the controls and ALS patients’iPSCs.Results:The iPSCs from the two FALS patients were capable of differentiation into MNs carrying different SOD1 mutations and differentially expressed MN markers.We detected high SOD1 protein expression and high intracellular calcium levels in both the MN and iPSCs that were derived from the twoSOD1 mutant patients.However,at no time did we observe stronger LDH activity in the patient lines compared with the control lines.Conclusions:MNs derived from patient-specific iPSC lines can recapitulate key aspects of ALS pathogenesis,providing a cell-based disease model to further elucidate disease pathogenesis and explore gene repair coupled with cell-replacement therapy.Incremental mutant expressions of SOD1 in MNs may have disrupted MN function,either causing or contributing to the intracellular calcium disturbances,which could lead to the occurrence and development of the disease.展开更多
观察运动疲劳大鼠黑质致密区(substantia nigra zonacompacta,SNc)多巴胺(dopamine,DA)神经元自发放电特征,探讨运动疲劳产生的中枢机制。方法:采用胞外玻璃微电极技术,在体观察运动疲劳后大鼠SNc区DA神经元自发电活动的变化。结果:运...观察运动疲劳大鼠黑质致密区(substantia nigra zonacompacta,SNc)多巴胺(dopamine,DA)神经元自发放电特征,探讨运动疲劳产生的中枢机制。方法:采用胞外玻璃微电极技术,在体观察运动疲劳后大鼠SNc区DA神经元自发电活动的变化。结果:运动疲劳大鼠SNc区DA能神经元自发单放电频率较对照组显著降低,神经元出现了不规则放电,且爆发式放电比例明显增多,放电间隔直方图成正偏态或随机分布(AI<1),ISI和CV值均显著大于对照组。结论:运动疲劳大鼠SNc区DA能神经元电活动出现明显改变,主要特征为兴奋性和活动规律性降低。SNc和纹状体的腹外侧和背外侧区构成的黑质—纹状体DA能神经通路参与了基底神经节对运动的调节,也是运动疲劳调控的重要中枢脑区之一。展开更多
基金supported by the“Double First-Class”University Project(CPU2022QZ31,China)。
文摘Both cholinergic dysfunction and protein citrullination are the hallmarks of rheumatoid arthritis(RA),but the relationship between the two phenomena remains unclear.We explored whether and how cholinergic dysfunction accelerates protein citrullination and consequently drives the development of RA.Cholinergic function and protein citrullination levels in patients with RA and collageninduced arthritis(CIA)mice were collected.In both neuron-macrophage coculture system and CIA mice,the effect of cholinergic dysfunction on protein citrullination and expression of peptidylarginine deiminases(PADs)was assessed by immunofluorescence.The key transcription factors for PAD4 expression were predicted and validated.Cholinergic dysfunction in the patients with RA and CIA mice negatively correlated with the degree of protein citrullination in synovial tissues.The cholinergic or alpha7 nicotinic acetylcholine receptor(a7nAChR)deactivation and activation resulted in the promotion and reduction of protein citrullination in vitro and in vivo,respectively.Especially,the activation deficiency of a7nAChR induced the earlier onset and aggravation of CIA.Furthermore,deactivation of a7nAChR increased the expression of PAD4 and specificity protein-3(SP3)in vitro and in vivo.Our results suggest that cholinergic dysfunction-induced deficient a7nAChR activation,which induces the expression of SP3 and its downstream molecule PAD4,accelerating protein citrullination and the development of RA.
基金by National Natural Science Foundation of China(NSFC 81271251)The Science and Technology Developmental Fund of Shandong Province,China(2012GGA15049).
文摘Parkinson’s disease(PD)is one of the most common neurodegenerative disorders of aging,characterized by the degeneration of dopamine neurons(DA neurons)in the substantial nigra,leading to the advent of both motor symptoms and non-motor symptoms.Current treatments include electrical stimulation of the affected brain areas and dopamine replacement therapy.Even though both categories are effective in treating PD patients,the disease progression cannot be stopped.The research advance into cell therapies provides exciting potential for the treatment of PD.Current cell sources include neural stem cells(NSCs)from fetal brain tissues,human embryonic stem cells(hESCs),induced pluripotent stem cells(iPSCs)and directly induced dopamine neurons(iDA neurons).Here,we evaluate the research progress in different cell sources with a focus on using iPSCs as a valuable source and propose key challenges for developing cells suitable for large-scale clinical applications in the treatment of PD.
基金supported by Sanofi-Aventis,The Helmsley Charitable Trust and The Ellison Medical Foundationsupported by"Thousand Young Talents"program of China+3 种基金National Laboratory of Biomacromolecules,Strategic Priority Research Program of the Chinese Academy of Sciencesa CIRM training grant fellowship(No.TG2-01158)a Glenn foundation grantpartially supported by an AFAR/Ellison Medical Foundation postdoctoral fellowship.
文摘The combination of disease-specific human induced pluripotent stem cells(iPSC)and directed cell differentiation offers an ideal platform for modeling and studying many inherited human diseases.Wilson’s disease(WD)is a monogenic disorder of toxic copper accumulation caused by pathologic mutations of the ATP7B gene.WD affects multiple organs with primary manifestations in the liver and central nervous system(CNS).In order to better investigate the cellular pathogenesis of WD and to develop novel therapies against various WD syndromes,we sought to establish a comprehensive platform to differentiate WD patient iPSC into both hepatic and neural lineages.Here we report the generation of patient iPSC bearing a Caucasian population hotspot mutation of ATP7B.Combining with directed cell differentiation strategies,we successfully differentiated WD iPSC into hepatocyte-like cells,neural stem cells and neurons.Gene expression analysis and cDNA sequencing confirmed the expression of the mutant ATP7B gene in all differentiated cells.Hence we established a platform for studying both hepatic and neural abnormalities of WD,which may provide a new tool for tissue-specific disease modeling and drug screening in the future.
基金financially supported by the National Science Foundation of China(nos.21788102,51873092,and 51961160730)the Research Grants Council of Hong Kong(nos.16305518,16305618,N-HKUST609/19,AHKUST605/16,and C6009-17G)+2 种基金the Innovation and Technology Commission(nos.ITC-CNERC14SC01 and ITCPD/17-9)Ming Wai Lau Centre for Reparative Medicine Associate Member Program(no.MWLC19SC02)the National Key Research and Development Program of China(no.2017YFE0132200).
文摘Oxidative damage to cells leads to accumulated harmful wastes,which in turn aggravate the imbalance of reactive oxygen species(ROS)and related diseases.Therefore,provoking the cellular defense system against severe oxidation and maintaining ROS homeostasis are desired.Herein,we designed and synthesized a powerful mitochondria-targeting aggregation-induced emission photosensitizer(named DTCSPY)by maximal restriction of heat dissipation.It is demonstrated that taming ROS generation within mitochondria through photosensitization-triggered autophagy via DTCSPY achieved a better neuroprotective effect against oxidative damages than Nacety-L-cysteine and vitamin C.This work not only provides a new way to design high-performance photosensitizers by regulating the photophysical property,but also verifies the concept that taming ROS can be used for cell protection against destructive oxidation,thereby displaying broad prospects for alleviating oxidation-related diseases and promoting cell-based therapy.
基金supported by FIS PI16/00786 grant,Instituto de Salud Carlos Ⅲ,Spain and Fondo Europeo de Desarrollo Regional(FEDER-Unión Europea),Proyectos de Investigación de Excelencia de la Junta de Andalucía CTS-5725AEPMI(Asociación de Enfermos de Patología Mitocondrial) and ENACH(Asociación de Enfermos de Neurodegeneración con Acumulación Cerebral de Hierro)(to JASA)
文摘Neurodegeneration with brain iron accumulation is a broad term that describes a heterogeneous group of progressive and invalidating neurologic disorders in which iron deposits in certain brain areas,mainly the basal ganglia.The predominant clinical symptoms include spasticity,progressive dystonia,Parkinson's disease-like symptoms,neuropsychiatric alterations,and retinal degeneration.Among the neurodegeneration with brain iron accumulation disorders,the most frequent subtype is pantothenate kinase-associated neurodegeneration(PKAN) caused by defects in the gene encoding the enzyme pantothenate kinase 2(PANK2)which catalyzed the first reaction of the coenzyme A biosynthesis pathway.Currently there is no effective treatment to prevent the inexorable course of these disorders.The aim of this review is to open up a discussion on the utility of using cellular models derived from patients as a valuable tool for the development of precision medicine in PKAN.Recently,we have described that dermal fibroblasts obtained from PKAN patients can manifest the main pathological changes of the disease such as intracellular iron accumulation accompanied by large amounts of lipofuscin granules,mitochondrial dysfunction and a pronounced increase of markers of oxidative stress.In addition,PKAN fibroblasts showed a morphological senescence-like phenotype.Interestingly,pantothenate supplementation,the substrate of the PANK2 enzyme,corrected all pathophysiological alterations in responder PKAN fibroblasts with low/residual PANK2 enzyme expression.However,pantothenate treatment had no favourable effect on PKAN fibroblasts harbouring mutations associated with the expression of a truncated/incomplete protein.The correction of pathological alterations by pantothenate in individual mutations was also verified in induced neurons obtained by direct reprograming of PKAN fibroblasts.Our observations indicate that pantothenate supplementation can increase/stabilize the expression levels of PANK2 in specific mutations.Fibroblasts and induced neuron
基金Ministry of Science and ICT,and Ministry of Health and Welfare(2021M3E5E5096464,Republic of Korea)Basic Science Research Program of the National Research Foundation of Korea(NRF)funded by the Ministry of Education(NRF-2022R1A6A1A03053343).
文摘Background: The ε4 allele of apolipoprotein E (APOE ε4) is the strongest known genetic risk factor for late-onset Alzheimer’s disease (AD), associated with amyloid pathogenesis. However, it is not clear how APOE ε4 accelerates amyloid-beta (Aβ) deposition during the seeding stage of amyloid development in AD patient neurons. Methods: AD patient induced neurons (iNs) with an APOE ε4 inducible system were prepared from skin fibroblasts of AD patients. Transcriptome analysis was performed using RNA isolated from the AD patient iNs expressing APOE ε4 at amyloid-seeding and amyloid-aggregation stages. Knockdown of IGFBP3 was applied in the iNs to investigate the role of IGFBP3 in the APOE ε4-mediated amyloidosis. Results: We optimized amyloid seeding stage in the iNs of AD patients that transiently expressed APOE ε4. Remarka-bly, we demonstrated that Aβ pathology was aggravated by the induction of APOE ε4 gene expression at the amyloid early-seeding stage in the iNs of AD patients. Moreover, transcriptome analysis in the early-seeding stage revealed that IGFBP3 was functionally important in the molecular pathology of APOE ε4-associated AD. Conclusions: Our findings suggest that the presence of APOE ε4 at the early Aβ-seeding stage in patient iNs is critical for aggravation of sporadic AD pathology. These results provide insights into the importance of APOE ε4 expression for the progression and pathogenesis of sporadic AD.
文摘In vitro studies have demonstrated that many factors of bone morphogenetic proteins （BMPs） induce cholinergic differentiation of neural stem cells. However, BMP retains the potential to induce increased numbers of cholinergic neurons in central nervous system regions that are rich in cholinergic cells, which is an important determinant of BMP. Therefore, BMP-4 was added to neural stem cell culture medium or the adult rat hippocampal dentate gyrus. Results demonstrated that BMP-4 induced cholinergic differentiation of neural stem cells in vitro and increased the number of cholinergic neurons in the adult rat hippocampal dentate gyrus.
基金supported by grants from National Natural Science Foundation of China(No.31670987)Beijing Science Foundation(No.7192223)。
文摘Background:Investigations of the pathogenic mechanisms in motor neurons(MNs)derived from amyotrophic lateral sclerosis(ALS)disease-specific induced pluripotent stem(iPS)cell lines could improve understanding of the issues affecting MNs.Therefore,in this study we explored mutant superoxide dismutase 1(SOD1)protein expression in MNs derived from the iPS cell lines of ALS patients carrying different SOD1 mutations.Methods:We generated induced pluripotent stem cell(iPSC)lines from two familial ALS(FALS)patients withSOD1-V14M andSOD1-C111Y mutations,and then differentiated them into MNs.We investigated levels of the SOD1 protein in iPSCs and MNs,the intracellular Ca2+levels in MNs,and the lactate dehydrogenase(LDH)activity in the process of differentiation into the MNs derived from the controls and ALS patients’iPSCs.Results:The iPSCs from the two FALS patients were capable of differentiation into MNs carrying different SOD1 mutations and differentially expressed MN markers.We detected high SOD1 protein expression and high intracellular calcium levels in both the MN and iPSCs that were derived from the twoSOD1 mutant patients.However,at no time did we observe stronger LDH activity in the patient lines compared with the control lines.Conclusions:MNs derived from patient-specific iPSC lines can recapitulate key aspects of ALS pathogenesis,providing a cell-based disease model to further elucidate disease pathogenesis and explore gene repair coupled with cell-replacement therapy.Incremental mutant expressions of SOD1 in MNs may have disrupted MN function,either causing or contributing to the intracellular calcium disturbances,which could lead to the occurrence and development of the disease.
文摘观察运动疲劳大鼠黑质致密区(substantia nigra zonacompacta,SNc)多巴胺(dopamine,DA)神经元自发放电特征,探讨运动疲劳产生的中枢机制。方法:采用胞外玻璃微电极技术,在体观察运动疲劳后大鼠SNc区DA神经元自发电活动的变化。结果:运动疲劳大鼠SNc区DA能神经元自发单放电频率较对照组显著降低,神经元出现了不规则放电,且爆发式放电比例明显增多,放电间隔直方图成正偏态或随机分布(AI<1),ISI和CV值均显著大于对照组。结论:运动疲劳大鼠SNc区DA能神经元电活动出现明显改变,主要特征为兴奋性和活动规律性降低。SNc和纹状体的腹外侧和背外侧区构成的黑质—纹状体DA能神经通路参与了基底神经节对运动的调节,也是运动疲劳调控的重要中枢脑区之一。
