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Magnetic resonance imaging focused on the ferritin heavy chain 1 reporter gene detects neuronal differentiation in stem cells

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Neural Regeneration Research 2023年第7期18卷 1563-1569页

作者： Xiao-Ya He Yi-Rui Zhou Tong Mu Yi-Fan Liao Li Jiang Yong Qin Jin-Hua Cai Department of Radiology Children’s Hospital of Chongqing Medical UniversityNational Clinical Research Center for Child Health and DisordersMinistry of Education Key Laboratory of Child Development and DisordersChongqing Key Laboratory of PediatricsChongqingChina Department of Endocrinology and Metabolism West China HospitalSichuan UniversityChengduSichuan ProvinceChina Department of Nuclear Medicine The Second Hospital of the Army Medical UniversityChongqingChina Department of Neurology Children’s Hospital of Chongqing Medical UniversityChongqingChina

The neuronal differentiation of mesenchymal stem cells offers a new strategy for the treatment of neurological disorders.Thus,there is a need to identify a noninvasive and sensitive in vivo imaging approach for real-time monitoring of transplanted stem cells.Our previous study confirmed that magnetic resonance imaging,with a focus on the ferritin heavy chain 1 reporter gene,could track the proliferation and differentiation of bone marrow mesenchymal stem cells that had been transduced with lentivirus carrying the ferritin heavy chain 1 reporter gene.However,we could not determine whether or when bone marrow mesenchymal stem cells had undergone neuronal differentiation based on changes in the magnetic resonance imaging signal.To solve this problem,we identified a neuron-specific enolase that can be differentially expressed before and after neuronal differentiation in stem cells.In this study,we successfully constructed a lentivirus carrying the neuron-specific enolase promoter and expressing the ferritin heavy chain 1 reporter gene;we used this lentivirus to transduce bone marrow mesenchymal stem cells.Cellular and animal studies showed that the neuron-specific enolase promoter effectively drove the expression of ferritin heavy chain 1 after neuronal differentiation of bone marrow mesenchymal stem cells;this led to intracellular accumulation of iron and corresponding changes in the magnetic resonance imaging signal.In summary,we established an innovative magnetic resonance imaging approach focused on the induction of reporter gene expression by a neuron-specific promoter.This imaging method can be used to noninvasively and sensitively detect neuronal differentiation in stem cells,which may be useful in stem cell-based therapies.

关键词： ferric ammonium citrate ferritin heavy chain 1 gene magnetic resonance imaging mesenchymal stem cells molecular imaging neuronal differentiation neuron-like cells neurons neuron-specific enolase promoter

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Presenilin mutations and their impact on neuronal differentiation in Alzheimer’s disease

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Neural Regeneration Research 2022年第1期17卷 31-37页

作者： Mercedes A.Hernández-Sapiéns Edwin E.Reza-Zaldívar Ana L.Márquez-Aguirre Ulises Gómez-Pinedo Jorge Matias-Guiu Ricardo R.Cevallos Juan C.Mateos-Díaz Víctor J.Sánchez-González Alejandro A.Canales-Aguirre Unidad de Evaluación Preclínica Unidad de Biotecnología Médica y FarmacéuticaCentro de Investigación y Asistencia en Tecnología y Diseño del Estado de JaliscoGuadalajaraMéxico Instituto de Neurociencias IdISSCHospital Clínico San CarlosMadridEspaña Biochemistry and Molecular Genetics Department University of AlabamaBirminghamAlabama Unidad de Biotecnología Industrial Centro de Investigación y Asistencia en Tecnología y Diseño del Estado de JaliscoGuadalajaraMéxico Centro Universitario de los Altos Universidad de GuadalajaraGuadalajaraMéxico

The presenilin genes(PSEN1 and PSEN2)are mainly responsible for causing early-onset familial Alzheimer’s disease,harboring~300 causative mutations,and representing~90%of all mutations associated with a very aggressive disease form.Presenilin 1 is the catalytic core of theγ-secretase complex that conducts the intramembranous proteolytic excision of multiple transmembrane proteins like the amyloid precursor protein,Notch-1,N-and E-cadherin,LRP,Syndecan,Delta,Jagged,CD44,ErbB4,and Nectin1a.Presenilin 1 plays an essential role in neural progenitor maintenance,neurogenesis,neurite outgrowth,synaptic function,neuronal function,myelination,and plasticity.Therefore,an imbalance caused by mutations in presenilin 1/γ-secretase might cause aberrant signaling,synaptic dysfunction,memory impairment,and increased Aβ42/Aβ40 ratio,contributing to neurodegeneration during the initial stages of Alzheimer’s disease pathogenesis.This review focuses on the neuronal differentiation dysregulation mediated by PSEN1 mutations in Alzheimer’s disease.Furthermore,we emphasize the importance of Alzheimer’s disease-induced pluripotent stem cells models in analyzing PSEN1 mutations implication over the early stages of the Alzheimer’s disease pathogenesis throughout neuronal differentiation impairment.

关键词： familial Alzheimer’s disease familial Alzheimer’s disease-induced pluripotent stem cells models induced pluripotent stem cells neurogenesis neuronal differentiation Notch presenilin 1 PSEN1 mutations γ-secretase complex

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Reducing host aldose reductase activity promotes neuronal differentiation of transplanted neural stem cells at spinal cord injury sites and facilitates locomotion recovery

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Neural Regeneration Research 2022年第8期17卷 1814-1820页

作者： Kun Zhang Wen-Can Lu Ming Zhang Qian Zhang Pan-Pan Xian Fang-Fang Liu Zhi-Yang Chen Chung Sookja Kim Sheng-Xi Wu Hui-Ren Tao Ya-Zhou Wang Department of Neurobiology and Institute of Neurosciences School of Basic MedicineFourth Military Medical UniversityXi’anShaanxi ProvinceChina Key Laboratory of Spine and Spinal Cord Injury Repair and Regeneration Tongji UniversityShanghaiChina Department of Spine Surgery Shenzhen University General HospitalShenzhenGuangdong ProvinceChina Department of Neurology Hainan Hospital of Chinese PLA General HospitalSanyaHainan ProvinceChina Department of Anesthesiology Tangdu HospitalFourth Military Medical UniversityXi’anShaanxi ProvinceChina Faculty of Medicine Macao University of Science and TechnologyMacao Special Administrative RegionChina

Neural stem cell(NSC)transplantation is a promising strategy for replacing lost neurons following spinal cord injury.However,the survival and differentiation of transplanted NSCs is limited,possibly owing to the neurotoxic inflammatory microenvironment.Because of the important role of glucose metabolism in M1/M2 polarization of microglia/macrophages,we hypothesized that altering the phenotype of microglia/macrophages by regulating the activity of aldose reductase(AR),a key enzyme in the polyol pathway of glucose metabolism,would provide a more beneficial microenvironment for NSC survival and differentiation.Here,we reveal that inhibition of host AR promoted the polarization of microglia/macrophages toward the M2 phenotype in lesioned spinal cord injuries.M2 macrophages promoted the differentiation of NSCs into neurons in vitro.Transplantation of NSCs into injured spinal cords either deficient in AR or treated with the AR inhibitor sorbinil promoted the survival and neuronal differentiation of NSCs at the injured spinal cord site and contributed to locomotor functional recovery.Our findings suggest that inhibition of host AR activity is beneficial in enhancing the survival and neuronal differentiation of transplanted NSCs and shows potential as a treatment of spinal cord injury.

关键词： aldose reductase functional recovery inflammation macrophage microglia neural stem cell transplantation neuronal differentiation polarization spinal cord injury

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Total Ginsenoside Extract from Panax ginseng Enhances Neural Stem Cell Proliferation and neuronal differentiation by Inactivating GSK-3β

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Chinese Journal of Integrative Medicine 2022年第3期28卷 229-235页

作者： LIN Kai-li ZHANG Ji CHUNG Hau-lam WU Xin-yi LIU Bin ZHAO Bo-xin SZE Stephen Cho-wing ZHOU Ping-zheng YUNG Ken Kin-lam ZHANG Shi-qing School of Public Health Guangzhou Medical UniversityGuangzhou(511436)China Department of Biology Hong Kong Baptist University(HKBU)Hong Kong Special Administrative Region(999077)China HKBU Shenzhen Research Institute and Continuing Education ShenzhenGuangdong Provinee(518057)China Department of Plastic Surgery Zhujiang HospitalSouthern Medical UniversityGuangzhou(510515)China Guangzhou Institute of Cardiovascular Disease the Second Affiliated Hospital of Guangzhou Medical UniversityGuangzhou(510260)China Department of Pharmacy Nan fang HospitalSouther n Medical Un iversityGuan gzhou(510515)China Guangdong Provincial Key Laboratory of New Drug Screening School of Pharmaceutical SciencesSouthern Medical UniversityGuangzhou(510515)China

Objective:To study the effects of total ginsenosides(TG)extract from Panax ginseng on neural stem cell(NSC)proliferation and differentiation and their underlying mechanisms.Methods:The migration of NSCs after treatment with various concentrations of TG extract(50,100,or 200μg/mL)were monitored.The proliferation of NSCs was examined by a combination of cell counting kit-8 and neurosphere assays.NSC differentiation mediated by TG extract was evaluated by Western blotting and immunofluorescence staining to monitor the expression of nestin and microtubule associated protein 2(MAP2).The GSK-3β/β-catenin pathway in TG-treated NSCs was examined by Western blot assay.The NSCs with constitutively active GSK-3βmutant were made by adenovirus-mediated gene transfection,then the proliferation and differentiation of NSCs mediated by TG were further verified.Results:TG treatment significantly enhanced NSC migration(Pdifferentiation(Pneuronal differentiation by inactivating GSK-3β.

关键词： ginsenoside neural stem cell proliferation neuronal differentiation GSK-3β

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Cell cycle exit and neuronal differentiation 1-engineered embryonic neural stem cells enhance neuronal differentiation and neurobehavioral recovery after experimental traumatic brain injury

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Neural Regeneration Research 2022年第1期17卷 130-136页

作者： Ren Wang Dian-Xu Yang Ying-Liang Liu Jun Ding Yan Guo Wan-Hai Ding Heng-Li Tian Fang Yuan Department of Neurosurgery Shanghai Jiao Tong University Affiliated Sixth People’s HospitalSchool of MedicineShanghai Jiao Tong UniversityShanghaiChina Department of Neurosurgery Shanghai Tenth People’s HospitalTongji UniversityShanghaiChina

Our previous study showed that cell cycle exit and neuronal differentiation 1(CEND1)may participate in neural stem cell cycle exit and oriented differentiation.However,whether CEND1-transfected neural stem cells can improve the prognosis of traumatic brain injury remained unclear.In this study,we performed quantitative proteomic analysis and found that after traumatic brain injury,CEND1 expression was downregulated in mouse brain tissue.Three days after traumatic brain injury,we transplanted CEND1-transfected neural stem cells into the area surrounding the injury site.We found that at 5 weeks after traumatic brain injury,transplantation of CEND1-transfected neural stem cells markedly alleviated brain atrophy and greatly improved neurological function.In vivo and in vitro results indicate that CEND1 overexpression inhibited the proliferation of neural stem cells,but significantly promoted their neuronal differentiation.Additionally,CEND1 overexpression reduced protein levels of Notch1 and cyclin D1,but increased levels of p21 in CEND1-transfected neural stem cells.Treatment with CEND1-transfected neural stem cells was superior to similar treatment without CEND1 transfection.These findings suggest that transplantation of CEND1-transfected neural stem cells is a promising cell therapy for traumatic brain injury.This study was approved by the Animal Ethics Committee of the School of Biomedical Engineering of Shanghai Jiao Tong University,China(approval No.2016034)on November 25,2016.

关键词： cell cycle exit and neuronal differentiation 1 cyclin D1 embryonic neural stem cells neuronal differentiation genetic engineering overexpression mice Notch1 p21 traumatic brain injury

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Metabolic and proteostatic differences in quiescent and active neural stem cells

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Neural Regeneration Research 2024年第1期19卷 43-48页

作者： Jiacheng Yu Gang Chen Hua Zhu Yi Zhong Zhenxing Yang Zhihong Jian Xiaoxing Xiong Department of Neurosurgery Renmin Hospital of Wuhan UniversityWuhanHubei ProvinceChina

Adult neural stem cells are neurogenesis progenitor cells that play an important role in neurogenesis.Therefore,neural regeneration may be a promising target for treatment of many neurological illnesses.The regenerative capacity of adult neural stem cells can be chara cterized by two states:quiescent and active.Quiescent adult neural stem cells are more stable and guarantee the quantity and quality of the adult neural stem cell pool.Active adult neural stem cells are chara cterized by rapid proliferation and differentiation into neurons which allow for integration into neural circuits.This review focuses on diffe rences between quiescent and active adult neural stem cells in nutrition metabolism and protein homeostasis.Furthermore,we discuss the physiological significance and underlying advantages of these diffe rences.Due to the limited number of adult neural stem cells studies,we refe rred to studies of embryonic adult neural stem cells or non-mammalian adult neural stem cells to evaluate specific mechanisms.

关键词： adult neurogenesis cell metabolic pathway cellular proliferation neural stem cell niches neural stem cells neuronal differentiation nutrient sensing pathway proteostasis

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Resveratrol promotes the survival and neuronal differentiation of hypoxia-conditioned neuronal progenitor cells in rats with cerebral ischemia

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Frontiers of Medicine 2021年第3期15卷 472-485页

作者： Yao Yao Rui Zhou Rui Bai Jing Wang Mengjiao Tu Jingjing Shi Xiao He Jinyun Zhou Liu Feng Yuanxue Gao Fahuan Song Feng Lan Xingguo Liu Mei Tian Hong Zhang Department of Nuclear Medicine and PET-CT Center The Second Hospital of Zhejiang University School of MedicineHangzhou 310009China Beijing Laboratory for Cardiovascular Precision Medicine Beijing Anzhen HospitalCapital Medical UniversityBeijing 100029China Key Laboratory for Biomedical Engineering of Ministry of Education Zhejiang UniversityHangzhou 310027China Guangzhou Regenerative Medicine and Health Guangdong Laboratory Guangzhou Institutes of Biomedicine and HealthChinese Academy of SciencesGuangzhou 510530China Shanxi Medical University Taiyuan 030001China The College of Biomedical Engineering and Instrument Science of Zhejiang University Hangzhou 310027China

Hypoxia conditioning could increase the survival of transplanted neuronal progenitor cells(NPCs)in rats with cerebral ischemia but could also hinder neuronal differentiation partly by suppressing mitochondrial metabolism.In this work,the mitochondrial metabolism of hypoxia-conditioned NPCs(hcNPCs)was upregulated via the additional administration of resveratrol,an herbal compound,to resolve the limitation of hypoxia conditioning on neuronal differentiation.Resveratrol was first applied during the in vitro neuronal differentiation of hcNPCs and concurrently promoted the differentiation,synaptogenesis,and functional development of neurons derived from hcNPCs and restored the mitochondrial metabolism.Furthermore,this herbal compound was used as an adjuvant during hcNPC transplantation in a photothrombotic stroke rat model.Resveratrol promoted neuronal differentiation and increased the long-term survival of transplanted hcNPCs.18-fluorine fluorodeoxyglucose positron emission tomography and rotarod test showed that resveratrol and hcNPC transplantation synergistically improved the neurological and metabolic recovery of stroke rats.In conclusion,resveratrol promoted the neuronal differentiation and therapeutic efficiency of hcNPCs in stroke rats via restoring mitochondrial metabolism.This work suggested a novel approach to promote the clinical translation of NPC transplantation therapy.

关键词： neuronal progenitor cells resveratrol cerebral ischemia neuronal differentiation mitochondrial metabolism positron emission tomography

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Non-catalytic roles for TET1 protein negatively regulating neuronal differentiation through srGAP3 in neuroblastoma cells

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Protein & Cell 2016年第5期7卷 351-361页

作者： Jie Gao Yue Ma Hua-Lin Fu Qian Luo ZhenWang Yu-Huan Xiao Hao Yang Da-Xiang Cui Wei-Lin Jin School of Life Sciences and Biotechnology Shanghai Jiao Tong University Shanghai 200240 China institute of Nano Biomedicine and Engineering Department of Instrument Science and Engineering School of Electronic Information and Electronic Engineering Shanghai Jiao Tong University Shanghai 200240 China National Center for Translational Medicine Shanghai Jiao Tong University Shanghai 200240 China Department of Experimental Surgery Tangdu Hospital Fourth Military Medical University Xi'an 710038 China Clinical Stem Cell Research Center Renji Hospital School of Medicine Shanghai Jiao Tong University Shanghai 200127 China

The methylcytosine dioxygenases TET proteins （TET1, TET2, and TET3） play important regulatory roles in neural function. In this study, we investigated the role of TET proteins in neuronal differentiation using Neuro2a cells as a model. We observed that knockdown of TET1, TET2 or TET3 promoted neuronal differentiation of Neuro2a cells, and their overexpression inhibited VPA （valproic acid）-induced neuronal differentiation, suggesting all three TET proteins negatively regulate neu- ronal differentiation of Neuro2a cells. Interestingly, the inducing activity of TET protein is independent of its enzymatic activity. Our previous studies have demon- strated that srGAP3 can negatively regulate neuronal differentiation of Neuro2a cells. Furthermore, we revealed that TET1 could positively regulate srGAP3 expression independent of its catalytic activity, and srGAP3 is required for TET-mediated neuronal differentiation of Neuro2a cells. The results presented here may facilitate better understanding of the role of TET proteins in neuronal differentiation, and provide a possible therapy target for neuroblastoma.

关键词： methylcytosine dioxygenase TET1,srGAP3 neuronal differentiation neuroblastoma cells

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Integrated transcriptome analysis of human iPS cells derived from a fragile X syndrome patient during neuronal differentiation

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Science China(Life Sciences) 2016年第11期59卷 1093-1105页

作者： Ping Lu Xiaolong Chen Yun Feng Qiao Zeng Cizhong Jiang Xianmin Zhu Guoping Fan Zhigang Xue Tongji Stem Cell Research Center Tongji University School of MedicineShanghai 200092China Tongji University School of Life Sciences and TechnologyShanghai 200092China Department of Human Genetics David Geffen School of MedicineUniversity of California Los AngelesLos Angeles CA 90095USA Translational Center for Stem Cell Research Tongji HospitalDepartment of Regenera-tive MedicineTongji University School of MedicineShanghai 200065China Tongji University Suzhou Institute Suzhou 215101China

Fragile X syndrome(FXS) patients carry the expansion of over 200 CGG repeats at the promoter of fragile X mental retardation 1(FMR1), leading to decreased or absent expression of its encoded fragile X mental retardation protein(FMRP). However, the global transcriptional alteration by FMRP deficiency has not been well characterized at single nucleotide resolution, i.e., RNA-seq. Here,we performed in-vitro neuronal differentiation of human induced pluripotent stem(iPS) cells that were derived from fibroblasts of a FXS patient(FXS-iPSC). We then performed RNA-seq and examined the transcriptional misregulation at each intermediate stage during in-vitro differentiation of FXS-iPSC into neurons. After thoroughly analyzing the transcriptomic data and integrating them with those from other platforms, we found up-regulation of many genes encoding TFs for neuronal differentiation(WNT1, BMP4,POU3F4, TFAP2 C, and PAX3), down-regulation of potassium channels(KCNA1, KCNC3, KCNG2, KCNIP4, KCNJ3, KCNK9,and KCNT1) and altered temporal regulation of SHANK1 and NNAT in FXS-iPSC derived neurons, indicating impaired neuronal differentiation and function in FXS patients. In conclusion, we demonstrated that the FMRP deficiency in FXS patients has significant impact on the gene expression patterns during development, which will help to discover potential targeting candidates for the cure of FXS symptoms.

关键词： fragile X syndrome induced pluripotent stem cells neuronal differentiation transcriptome

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Reprogrammed mouse astrocytes retain a“memory”of tissue origin and possess more tendencies for neuronal differentiation than reprogrammed mouse embryonic fibroblasts

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Protein & Cell 2011年第2期2卷 128-140页

作者： Changhai Tian Yongxiang Wang Lijun Sun Kangmu Ma Jialin C.Zheng Laboratory of Neuroimmunology and Regenerative Therapy University of Nebraska Medical CenterOmahaNE 68198-5930USA Departments of Pharmacology/Experimental Neuroscience University of Nebraska Medical CenterOmahaNE 68198-5930USA Departments of Pathology/Microbiology University of Nebraska Medical CenterOmahaNE 68198-5930USA

Direct reprogramming of a variety of somatic cells with the transcription factors Oct4(also called Pou5f1),Sox2 with either Klf4 and Myc or Lin28 and Nanog generates the induced pluripotent stem cells(iPSCs)with marker similarity to embryonic stem cells.However,the difference between iPSCs derived from different origins is unclear.In this study,we hypothesized that reprogrammed cells retain a“memory”of their origins and possess additional potential of related tissue differentiation.We reprogrammed primary mouse astrocytes via ectopic retroviral expression of OCT3/4,Sox2,Klf4 and Myc and found the iPSCs from mouse astrocytes expressed stem cell markers and formed teratomas in SCID mice containing derivatives of all three germ layers similar to mouse embryonic stem cells besides semblable morphologies.To test our hypothesis,we compared embryonic bodies(EBs)formation and neuronal differentiation between iPSCs from mouse embryonic fibroblasts(MEFsiPSCs)and iPSCs from mouse astrocytes(mAsiPSCs).We found that mAsiPSCs grew slower and possessed more potential for neuronal differentiation compared to MEFsiPSCs.Our results suggest that mAsiPSCs retain a“memory”of the central nervous system,which confers additional potential upon neuronal differentiation.

关键词： mouse astrocytes induced pluripotent stem cells neural progenitor cells neuronal differentiation

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