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Frontiers of Medicine 2023年第6期17卷 1186-1203页

作者： Jie Shao Wenjuan Wang Baorui Tao Zihao Cai Haixia Li Jinhong Chen Department of General Surgery Huashan HospitalFudan UniversityShanghai 200040China Department of Pathology Huashan HospitalFudan UniversityShanghai 200040China

Through bioinformatics predictions,we identified that GTF2I and FAT1 were downregulated in thyroid carcinoma(TC).Further,Pearson’s correlation coefficient revealed a positive correlation between GTF2I expression and FAT1 expression.Therefore,we selected them for this present study,where the effects of bone marrow mesenchymal stem cell-derived EVs(BMSDs-EVs)enriched with GTF2I were evaluated on the epithelial–to–mesenchymal transition(EMT)and stemness maintenance in TC.The under-expression of GTF2I and FAT1 was validated in TC cell lines.Ectopically expressed GTF2I and FAT1 were found to augment malignant phenotypes of TC cells,EMT,and stemness maintenance.Mechanistic studies revealed that GTF2I bound to the promoter region of FAT1 and consequently upregulated its expression.MSC-EVs could shuttle GTF2I into TPC-1 cells,where GTF2I inhibited TC malignant phenotypes,EMT,and stemness maintenance by increasing the expression of FAT1 and facilitating the FAT1-mediated CDK4/FOXM1 downregulation.In vivo experiments confirmed that silencing of GTF2I accelerated tumor growth in nude mice.Taken together,our work suggests that GTF2I transferred by MSC-EVs confer antioncogenic effects through the FAT1/CDK4/FOXM1 axis and may be used as a promising biomarker for TC treatment.

关键词： thyroid carcinoma mesenchymal stem cell extracellular vesicle GTF2I FAT1 CDK4

Advances in extracellular vesicle functionalization strategies for tissue regeneration

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Bioactive Materials 2023年第7期25卷 500-526页

作者： Dandan Zheng Huitong Ruan Wei Chen Yuhui Zhang Wenguo Cui Hao Chen Hongxing Shen Department of Spine Surgery Renji HospitalShanghai JiaoTong University School of Medicine160 Pujian RoadShanghai200127PR China Department of Orthopaedics Shanghai Key Laboratory for Prevention and Treatment of Bone and Joint DiseasesShanghai Institute of Traumatology and OrthopaedicsRuijin HospitalShanghai Jiao Tong University School of Medicine197 Ruijin 2nd RoadShanghai200025China

extracellular vesicles(EVs)are nano-scale vesicles derived by cell secretion with unique advantages such as promoting cell proliferation,anti-inflammation,promoting blood vessels and regulating cell differentiation,which benefit their wide applications in regenerative medicine.However,the in vivo therapeutic effect of EVs still greatly restricted by several obstacles,including the off-targetability,rapid blood clearance,and undesired release.To address these issues,biomedical engineering techniques are vastly explored.This review summarizes different strategies to enhance EV functions from the perspective of drug loading,modification,and combination of biomaterials,and emphatically introduces the latest developments of functionalized EV-loaded biomaterials in different diseases,including cardio-vascular system diseases,osteochondral disorders,wound healing,nerve injuries.Challenges and future directions of EVs are also discussed.

关键词： extracellular vesicle Regenerative medicine Biomaterials Modification Drug delivery

Exercise promotes angiogenesis by enhancing endothelial cell fatty acid utilization via liver-derived extracellular vesicle miR-122-5p

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Journal of Sport and Health Science 2022年第4期11卷 495-508页

作者： Jing Lou Jie Wu Mengya Feng Xue Dang Guiling Wu Hongyan Yang Yan Wang Jia Li Yong Zhao Changhong Shi Jiankang Liu Lin Zhao Xing Zhang Feng Gao Center for Mitochondrial Biology and Medicine The Key Laboratory of Biomedical Information Engineering of Ministry of EducationSchool of Life Science and TechnologyXi’an Jiaotong UniversityXi’an 710049China School of Aerospace Medicine Fourth Military Medical UniversityXi’an 710032China Medical School of Chinese PLA Beijing 100853China Laboratory Animal Center Fourth Military Medical UniversityXi’an 710032China

Background:Angiogenesis constitutes a major mechanism responsible for exercise-induced beneficial effects.Our previous study identified a cluster of differentially expressed extracellular vesicle microRNAs(miRNAs)after exercise and found that some of them act as exerkines.However,whether these extracellular vesicle miRNAs mediate the exercise-induced angiogenesis remains unknown.Methods:A 9-day treadmill training was used as an exercise model in C57BL/6 mice.Liver-specific adeno-associated virus 8 was used to knock down microRNA-122-5p(miR-122-5p).Human umbilical vein endothelial cells were used in vitro.Results:Among these differentially expressed extracellular vesicle miRNAs,miR-122-5p was identified as a potent pro-angiogenic factor that activated vascular endothelial growth factor signaling and promoted angiogenesis both in vivo and in vitro.Exercise increased circulating levels of miR-122-5p,which was produced mainly by the liver and shuttled by extracellular vesicles in mice.Inhibition of circulating miR-122-5p or liver-specific knockdown of miR-122-5p significantly abolished the exercise-induced pro-angiogenic effect in skeletal muscles,and exerciseimproved muscle performance in mice.Mechanistically,miR-122-5p promoted angiogenesis through shifting substrate preference to fatty acids in endothelial cells,and miR-122-5p upregulated endothelial cell fatty-acid utilization by targeting 1-acyl-sn-glycerol-3-phosphate acyltransferase(AGPAT1).In addition,miR-122-5p increased capillary density in perilesional skin tissues and accelerated wound healing in mice.Conclusion:These findings demonstrated that exercise promotes angiogenesis through upregulation of liver-derived extracellular vesicle miR-122-5p,which enhances fatty acid utilization by targeting AGPAT1 in endothelial cells,highlighting the therapeutic potential of miR-122-5p in tissue repair.

关键词： Angiogenesis Endothelial cells Exerkine extracellular vesicle Metabolic shift

Intranasal administration of stem cell-derived exosomes for central nervous system diseases

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Neural Regeneration Research 2024年第6期19卷 1249-1255页

作者： Shuho Gotoh Masahito Kawabori Miki Fujimura Department of Neurosurgery Hokkaido University Graduate School of Medicine

Exosomes, lipid bilayer-enclosed small cellular vesicles, are actively secreted by various cells and play crucial roles in intercellular communication. These nanosized vesicles transport internalized proteins, mRNA, miRNA, and other bioactive molecules. Recent findings have provided compelling evidence that exosomes derived from stem cells hold great promise as a therapeutic modality for central nervous system disorders. These exosomes exhibit multifaceted properties including antiapoptotic, anti-inflammatory, neurogenic, and vasculogenic effects. Furthermore, exosomes offer several advantages over stem cell therapy, such as high preservation capacity, low immunogenicity, the ability to traverse the blood-brain barrier, and the potential for drug encapsulation. Consequently, researchers have turned their attention to exosomes as a novel therapeutic avenue. Nonetheless, akin to the limitations of stem cell treatment, the limited accumulation of exosomes in the injured brain poses a challenge to their clinical application. To overcome this hurdle, intranasal administration has emerged as a non-invasive and efficacious route for delivering drugs to the central nervous system. By exploiting the olfactory and trigeminal nerve axons, this approach enables the direct transport of therapeutics to the brain while bypassing the blood-brain barrier. Notably, exosomes, owing to their small size, can readily access the nerve pathways using this method. As a result, intranasal administration has gained increasing recognition as an optimal therapeutic strategy for exosomebased treatments. In this comprehensive review, we aim to provide an overview of both basic and clinical research studies investigating the intranasal administration of exosomes for the treatment of central nervous system diseases. Furthermore, we elucidate the underlying therapeutic mechanisms and offer insights into the prospect of this approach.

关键词： central nervous system disease exosome extracellular vesicle intranasal administration stem cell

Neural stem cells promote neuroplasticity: a promising therapeutic strategy for the treatment of Alzheimer’s disease

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Neural Regeneration Research 2024年第3期19卷 619-628页

作者： Jun Chang Yujiao Li Xiaoqian Shan Xi Chen Xuhe Yan Jianwei Liu Lan Zhao First Teaching Hospital of Tianjin University of Traditional Chinese Medicine TianjinChina National Clinical Research Center for Chinese Medicine Acupuncture and Moxibustion TianjinChina Tianjin University of Traditional Chinese Medicine TianjinChina

Recent studies have demonstrated that neuroplasticity,such as synaptic plasticity and neurogenesis,exists throughout the normal lifespan but declines with age and is significantly impaired in individuals with Alzheimer’s disease.Hence,promoting neuroplasticity may represent an effective strategy with which Alzheimer’s disease can be alleviated.Due to their significant ability to self-renew,differentiate,and migrate,neural stem cells play an essential role in reversing synaptic and neuronal damage,reducing the pathology of Alzheimer’s disease,including amyloid-β,tau protein,and neuroinflammation,and secreting neurotrophic factors and growth factors that are related to plasticity.These events can promote synaptic plasticity and neurogenesis to repair the microenvironment of the mammalian brain.Consequently,neural stem cells are considered to represent a potential regenerative therapy with which to improve Alzheimer’s disease and other neurodegenerative diseases.In this review,we discuss how neural stem cells regulate neuroplasticity and optimize their effects to enhance their potential for treating Alzheimer’s disease in the clinic.

关键词： Alzheimer’s disease amyloid-β cell therapy extracellular vesicle neural stem cell synaptic plasticity tau

Patient-derived induced pluripotent stem cells with a MERTK mutation exhibit cell junction abnormalities and aberrant cellular differentiation potential

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World Journal of Stem Cells 2024年第5期16卷 512-524页

作者： Hang Zhang Ling-Zi Wu Zhen-Yu Liu Zi-Bing Jin Beijing Institute of Ophthalmology Beijing Tongren Eye CenterBeijing Tongren HospitalCapital Medical UniversityBeijing 100730China

BACKGROUND Human induced pluripotent stem cell(hiPSC)technology is a valuable tool for generating patient-specific stem cells,facilitating disease modeling,and invest-igating disease mechanisms.However,iPSCs carrying specific mutations may limit their clinical applications due to certain inherent characteristics.AIM To investigate the impact of MERTK mutations on hiPSCs and determine whether hiPSC-derived extracellular vesicles(EVs)influence anomalous cell junction and differentiation potential.METHODS We employed a non-integrating reprogramming technique to generate peripheral blood-derived hiPSCs with and hiPSCs without a MERTK mutation.Chromo-somal karyotype analysis,flow cytometry,and immunofluorescent staining were utilized for hiPSC identification.Transcriptomics and proteomics were employed to elucidate the expression patterns associated with cell junction abnormalities and cellular differentiation potential.Additionally,EVs were isolated from the supernatant,and their RNA and protein cargos were examined to investigate the involvement of hiPSC-derived EVs in stem cell junction and differentiation.RESULTS The generated hiPSCs,both with and without a MERTK mutation,exhibited normal karyotype and expressed pluripotency markers;however,hiPSCs with a MERTK mutation demonstrated anomalous adhesion capability and differentiation potential,as confirmed by transcriptomic and proteomic profiling.Furthermore,hiPSC-derived EVs were involved in various biological processes,including cell junction and differentiation.CONCLUSION HiPSCs with a MERTK mutation displayed altered junction characteristics and aberrant differentiation potential.Furthermore,hiPSC-derived EVs played a regulatory role in various biological processes,including cell junction and differentiation.

关键词： Cell junction Cellular differentiation extracellular vesicle Human induced pluripotent stem cells Transcriptomics Proteomics

Pathogenic and therapeutic role of exosomes in neurodegenerative disorders

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Neural Regeneration Research 2024年第1期19卷 75-79页

作者： Christa C.Huber Hongmin Wang Division of Basic Biomedical Sciences Sanford School of MedicineUniversity of South DakotaVermillionSDUSA

Neurodegenerative disorders affect millions of people worldwide,and the prevalence of these disorders is only projected to rise as the number of people over 65 will drastically increase in the coming years.While therapies exist to aid in symptomatic relief,effective treatments that can stop or reve rse the progress of each neurodegenerative disease are lacking.Recently,research on the role of extracellular vesicles as disease markers and therapeutics has been intensively studied.Exosomes,30-150 nm in diameter,are one type of extracellular vesicles facilitating cell-to-cell communication.Exosomes are thought to play a role in disease propagation in a variety of neurodegenerative diseases,such as Alzheimer's disease,Parkinson s disease,and amyotrophic lateral sclerosis.Accordingly,the exosomes derived from the patients are an invaluable source of disease biomarkers.On the other hand,exosomes,especially those derived from stem cells,could serve as a therapeutic for these disorders,as seen by a rapid increase in clinical trials investigating the therapeutic efficacy of exosomes in different neurological diseases.This review summarizes the pathological burden and therapeutic approach of exosomes in neurodegenerative disorders.We also highlight how heat shock increases the yield of exosomes while still maintaining their therapeutic efficacy.Finally,this review concludes with outstanding questions that remain to be addressed in exosomal research.

关键词： aging Alzheimer's disease biomarker drug exosome extracellular vesicle neurodegenerative disease Parkinson's disease therapy

Microfluidics-based technologies for the analysis of extracellular vesicles at the single-cell level and single-vesicle level

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Chinese Chemical Letters 2022年第6期33卷 2893-2900页

作者： Fengjiao Zhu Yahui Ji Jiu Deng Linmei Li Xue Bai Xianming Liu Bingcheng Lin Yao Lu Department of Biotechnology Dalian Institute of Chemical PhysicsChinese Academy of SciencesDalian 116023China University of Chinese Academy of Sciences Beijing 100049China

extracellular vesicles(EVs)are membrane vesicles secreted by cells,playing critical roles in mediating intercellular communications for various physiological and pathological processes.Most of the EV analysis is currently performed at the bulk level,obscuring the origin of the EVs and diverse characteristics of the individual extracellular vesicle.Technologies to analyze the extracellular vesicles at the single-cell and single-vesicle levels are needed to evaluate EV comprehensively and decode the heterogeneity underlying EV secretion.Microfluidic platforms that could control and manipulate fluids at the microscale provide an efficient way to achieve the aims.Various microfluidics-based technologies are emerging to realize single-cell EV secretion analysis and single EV analysis,which would be summarized in this mini-review.

关键词： Microfluidics Single-cell EV analysis Single EV analysis extracellular vesicle Heterogeneity

Attenuation of amyotrophic lateral sclerosis via stem cell and extracellular vesicle therapy:An updated review

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Neuroprotection 2023年第2期1卷 130-138页

作者： Gavin Lockard Jonah Gordon Samantha Schimmel Bassel El Sayed Molly Monsour Svitlana Garbuzova-Davis Cesar V.Borlongan University of South Florida Morsani College of Medicine TampaFloridaUSA Department of Neurosurgery and Brain Repair Center of Excellence for Aging and Brain RepairUniversity of South Florida Morsani College of MedicineTampaFloridaUSA

Amyotrophic lateral sclerosis(ALS)is a rapidly fatal neurological disease characterized by upper and lower motor neuron degeneration.Though typically idiopathic,familial forms of ALS are commonly composed of a superoxide dismutase 1(SOD1)mutation.Basic science frequently utilizes SOD1 models in vitro and in vivo to replicate ALS conditions.Therapies are sparse;those that exist in the market extend life minimally,thus driving the demand for research to identify novel therapeutics.Transplantation of stem cells is a promising approach for many diseases and has shown efficacy in SOD1 models and clinical trials.The underlying mechanism for stem cell therapy presents an exciting venue for research investigations.Most notably,the paracrine actions of stem cell-derived extracellular vesicles(EVs)have been suggested as a potent mitigating factor.This literature review focuses on the most recent preclinical research investigating cellfree methods for treating ALS.Various avenues are being explored,differing on the EV contents(protein,microRNA,etc.)and on the cell target(astrocyte,endothelial cell,motor neuron-like cells,etc.),and both molecular and behavioral outcomes are being examined.Unfortunately,EVs may also play a role in propagating ALS pathology.Nonetheless,the overarching goal remains clear:to identify efficient cell-free techniques to attenuate the deadly consequences of ALS.

关键词： amyotrophic lateral sclerosis extracellular vesicle stem cell

Natural killer cell-derived extracellular vesicle significantly enhanced adoptive T cell therapy against solid tumors via versatilely immunomodulatory coordination

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Science China Chemistry 2021年第11期64卷 1999-2009页

作者： Weidong Nie Wenlin Fan Anqi Jiang Guanghao Wu Houli Liu Li-Li Huang Hai-Yan Xie School of Materials Science and Engineering Beijing Institute of TechnologyBeijing100081China School of Life Science Beijing Institute of TechnologyBeijing100081China Institute of Engineering Medicine Beijing Institute of TechnologyBeijing100081China

Insufficient tumor tropism,MHC classⅠmolecules(MHC-I)defects of tumor cells,and immunosuppressive tumor microenvironment(TME)seriously imperil the efficacy of adoptive T cell therapy on solid tumors.Here,natural killer cell-derived extracellular vesicle(Nev)is used as a versatile toolkit to synergistically improve adoptive T cell therapy for solid tumors.Specifically,Nev is modified with dibenzocyclooctynes(DBCO)linked with p H-sensitive benzoic-imine bonds;meanwhile,cytotoxic T lymphocyte(CTL)is decorated with azide groups.Then CTL is armed with Nev(CTL-Nev)through the click chemistry reaction.After systematic administration,Nev obviously promotes the tumor-targeting accumulation of CTL coming from its native tumor-tropism capability.Then,the cleavage of benzoic-imine bonds in the slightly acidic TME leads to the release of Nev,which not only directly induces tumor apoptosis but also promotes the action of CTL via multiplex pathways,such as up-regulating the MHC-I expression on tumor cells,reprogramming tumor-associated macrophages from pro-tumoral M2 phenotypes to tumoricidal M1 phenotypes.The all-around coordination of Nev with CTL results in potent tumor repression.

关键词： click chemistry adoptive T cell therapy natural killer cell extracellular vesicle tumor immunotherapy