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Mesenchymal stem cells,extracellular vesicles,and transcranial magnetic stimulation for ferroptosis after spinal cord injury

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Neural Regeneration Research 2023年第9期18卷 1861-1868页

作者： Qi-Feng Song Qian Cui Ya-Shi Wang Li-Xin Zhang Department of Rehabilitation Shengjing Hospital of China Medical UniversityShenyangLiaoning ProvinceChina

Spinal cord injury is characte rized by diffe rent aetiologies,complex pathogenesis,and diverse pathological changes.Current treatments are not ideal,and prognosis is generally poor.After spinal cord injury,neurons die due to various forms of cell death.Among them,fe rroptosis causes dysfunction after spinal cord injury,and no existing traditional treatments have been indicated to block its occurrence.Meanwhile,emerging therapies using mesenchymal stem cells,extracellular vesicles,and transcranial magnetic stimulation therapy are promising for reve rsing spinal co rd neuronal ferroptosis after spinal cord injury.However,no definitive studies have demonstrated the effectiveness of these approaches.This review summarizes the existing research on the mechanisms of ferroptosis;fe rroptosis after spinal cord injury;treatment of spinal cord injury with mesenchymal stem cells,extracellular vesicles,and transc ranial magnetic stimulation;and treatment of ferroptosis using mesenchymal stem cells,extracellular vesicles,and transc ranial magnetic stimulation.Inhibiting ferroptosis can promote the reversal of neurological dysfunction after spinal cord injury.In addition,mesenchymal stem cells,extracellular vesicles,and transc ranial magnetic stimulation can reve rse adverse outcomes of spinal cord injury and regulate ferroptosis-related fa ctors.Thus,it can be inferred that mesenchymal stem cells,extracellular vesicles,and transcranial magnetic stimulation have the potential to inhibit fe rroptosis after spinal cord injury.This review serves as a reference for future research to confirm these conclusions.

关键词： exosomes extracellular vesicles ferroptosis iron overload lipid peroxidation mesenchymal stem cells miRNAs spinal cord injury stem cells transcranial magnetic stimulation

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Neutralization of SARS-CoV-2 pseudovirus using ACE2-engineered extracellular vesicles

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Acta Pharmaceutica Sinica B 2022年第3期12卷 1523-1533,F0004页

作者： Canhao Wu Qin Xu Huiyuan Wang Bin Tu Jiaxin Zeng Pengfei Zhao Mingjie Shi Hong Qiu Yongzhuo Huang Artemisinin Research Center First Clinical SchoolGuangzhou University of Chinese MedicineGuangzhou 510450China State Key Laboratory of Drug Research Shanghai Institute of Materia MedicaChinese Academy of SciencesShanghai 201203China Zhongshan Institute for Drug Discovery SIMMCASZhongshan 528437China Center of Clinical Pharmacology The Second Affiliated Hospital of Zhejiang UniversitySchool of MedicineHangzhou 310009China NMPA Key Laboratory for Quality Research and Evaluation of Pharmaceutical Excipients Shanghai 201203China Taizhou University School of Advanced StudyInstitute of Natural Medicine and Health ProductTaizhou 318000China

The spread of coronavirus disease 2019(COVID-19)throughout the world has resulted in stressful healthcare burdens and global health crises.Developing an effective measure to protect people from infection is an urgent need.The blockage of interaction between angiotensin-converting enzyme 2(ACE2)and S protein is considered an essential target for anti-severe acute respiratory syndrome coronavirus 2(SARS-Co V-2)drugs.A full-length ACE2 protein could be a potential drug to block early entry of SARS-Co V-2 into host cells.In this study,a therapeutic strategy was developed by using extracellular vesicles(EVs)with decoy receptor ACE2 for neutralization of SARS-Co V-2.The EVs embedded with engineered ACE2(EVs-ACE2)were prepared;the EVs-ACE2 were derived from an engineered cell line with stable ACE2 expression.The potential effect of the EVs-ACE2 on anti-SARS-Co V-2 was demonstrated by both in vitro and in vivo neutralization experiments using the pseudovirus with the S protein(S-pseudovirus).EVs-ACE2 can inhibit the infection of S-pseudovirus in various cells,and importantly,the mice treated with intranasal administration of EVs-ACE2 can suppress the entry of S-pseudovirus into the mucosal epithelium.Therefore,the intranasal EVs-ACE2 could be a preventive medicine to protect from SARS-Co V-2 infection.This EVs-based strategy offers a potential route to COVID-19 drug development.

关键词： SARS-CoV-2 COVID-19 Spike protein Pseudovirus extracellular vesicles ACE2 Intranasal administration Neutralization

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Orange-derived extracellular vesicles nanodrugs for efficient treatment of ovarian cancer assisted by transcytosis effect

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Acta Pharmaceutica Sinica B 2023年第12期13卷 5121-5134页

作者： Feng Long Yao Pan Jinheng Li Suinan Sha Xiubo Shi Haoyan Guo Chuanqing Huang Qian Xiao Chao Fan Xingmei Zhang Jun-Bing Fan Ying Wang Cancer Research Institute Experimental Education/Administration CenterKey Laboratory of Functional Proteomics of Guangdong ProvinceSchool of Basic Medical SciencesSouthern Medical UniversityGuangzhou 510515China Department of Neurobiology School of Basic Medical SciencesSouthern Medical UniversityGuangzhou 510515China

extracellular vesicles(EVs)have recently received much attention about the application of drug carriers due to their desirable properties such as nano-size,biocompatibility,and high stability.Herein,we demonstrate orange-derived extracellular vesicles(OEV)nanodrugs(DN@OEV)by modifying cRGD-targeted doxorubicin(DOX)nanoparticles(DN)onto the surface of OEV,enabling significantly enhancing tumor accumulation and penetration,thereby efficiently inhibiting the growth of ovarian cancer.The obtained DN@OEV enabled to inducement of greater transcytosis capability in ovarian cancer cells,which presented the average above 10-fold transcytosis effect compared with individual DN.It was found that DN@OEV could trigger receptor-mediated endocytosis to promote early endosome/recycling endosomes pathway for exocytosis and simultaneously reduce degradation in the early endosomes-late endosomes-lysosome pathway,thereby inducing the enhanced transcytosis.In particular,the zombie mouse model bearing orthotopic ovarian cancer further validated DN@OEV presented high accumulation and penetration in tumor tissue by the transcytosis process.Our study indicated the strategy in enhancing transcytosis has significant implications for improving the therapeutic efficacy of thedrugdelivery system.

关键词： extracellular vesicles Transcytosis Cancer treatment Drug delivery system Targeted modification Receptor-mediated endocytosis Intracellular trafficking Recycling endosome

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Secretive derived from hypoxia preconditioned mesenchymal stem cells promote cartilage regeneration and mitigate joint inflammation via extracellular vesicles

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Bioactive Materials 2023年第9期27卷 98-112页

作者： Yanmeng Yang Yingnan Wu Dahou Yang Shu Hui Neo Nurul Dinah Kadir Doreen Goh Jian Xiong Tan Vinitha Denslin Eng Hin Lee Zheng Yang Department of Orthopaedic Surgery Yong Loo Lin School of MedicineNational University of SingaporeSingapore NUS Tissue Engineering Program Life Sciences InstituteNational University of SingaporeSingapore Critical Analytics for Manufacturing Personalised-Medicine Singapore-MIT Alliance for Research and TechnologySingapore

Secretome derived from mesenchymal stem cells (MSCs) have profound effects on tissue regeneration, which could become the basis of future MSCs therapies. Hypoxia, as the physiologic environment of MSCs, has great potential to enhance MSCs paracrine therapeutic effect. In our study, the paracrine effects of secretome derived from MSCs preconditioned in normoxia and hypoxia was compared through both in vitro functional assays and an in vivo rat osteochondral defect model. Specifically, the paracrine effect of total EVs were compared to that of soluble factors to characterize the predominant active components in the hypoxic secretome. We demonstrated that hypoxia conditioned medium, as well as the corresponding EVs, at a relatively low dosage, were efficient in promoting the repair of critical-sized osteochondral defects and mitigated the joint inflammation in a rat osteochondral defect model, relative to their normoxia counterpart. In vitro functional test shows enhancement through chondrocyte proliferation, migration, and matrix deposition, while inhibit IL-1β-induced chondrocytes senescence, inflammation, matrix degradation, and pro-inflammatory macrophage activity. Multiple functional proteins, as well as a change in EVs’ size profile, with enrichment of specific EV-miRNAs were detected with hypoxia preconditioning, implicating complex molecular pathways involved in hypoxia pre-conditioned MSCs secretome generated cartilage regeneration.

关键词： Hypoxia preconditioned MSCs Secretome extracellular vesicles Cartilage regeneration Joint inflammation

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Adipose mesenchymal stem cell-derived extracellular vesicles reduce glutamate-induced excitotoxicity in the retina

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Neural Regeneration Research 2023年第10期18卷 2315-2320页

作者： Tian-Qi Duan Zhao-Lin Gao Ai-Xiang Luo Dan Chen Jian-Bin Tong Ju-Fang Huang Department of Anatomy and Neurobiology School of Basic Medical SciencesCentral South UniversityChangshaHunan ProvinceChina Hunan Province Key Laboratory of Brain Homeostasis Third Xiangya HospitalCentral South UniversityChangshaHunan ProvinceChina

Adipose mesenchymal stem cells(ADSCs)have protective effects against glutamate-induced excitotoxicity,but ADSCs are limited in use for treatment of optic nerve injury.Studies have shown that the extracellular vesicles(EVs)secreted by ADSCs(ADSC-EVs)not only have the function of ADSCs,but also have unique advantages including non-immunogenicity,low probability of abnormal growth,and easy access to target cells.In the present study,we showed that intravitreal injection of ADSC-EVs substantially reduced glutamate-induced damage to retinal morphology and electroretinography.In addition,R28 cell pretreatment with ADSC-EVs before injury inhibited glutamate-induced overload of intracellular calcium,downregulation ofα-amino-3-hydroxy-5-methyl-4-isoxazoleproprionic acid receptor(AMPAR)subunit GluA2,and phosphorylation of GluA2 and protein kinase C alpha in vitro.A protein kinase C alpha agonist,12-O-tetradecanoylphorbol 13-acetate,inhibited the neuroprotective effects of ADSC-EVs on glutamate-induced R28 cells.These findings suggest that ADSCEVs ameliorate glutamate-induced excitotoxicity in the retina through inhibiting protein kinase C alpha activation.

关键词： adipose mesenchymal stem cells calcium overload electroretinography excitotoxicity extracellular vesicles GluA2 glutamate protein kinase C alpha R28 cells retina retinal ganglion cell

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New idea to promote the clinical applications of stem cells or their extracellular vesicles in central nervous system disorders:Combining with intranasal delivery

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Acta Pharmaceutica Sinica B 2022年第8期12卷 3215-3232页

作者： Yaosheng Li Honghui Wu Xinchi Jiang Yunfei Dong Juanjuan Zheng Jianqing Gao Institute of Pharmaceutics College of Pharmaceutical SciencesZhejiang UniversityHangzhou 310058China Dr.Li Dak Sum&Yip Yio Chin Center for Stem Cell and Regenerative Medicine Zhejiang UniversityHangzhou 310058China Hangzhou Institute of Innovative Medicine Zhejiang UniversityHangzhou 310058China

The clinical translation of stem cells and their extracellular vesicles(EVs)-based therapy for central nervous system(CNS) diseases is booming. Nevertheless, the insufficient CNS delivery and retention together with the invasiveness of current administration routes prevent stem cells or EVs from fully exerting their clinical therapeutic potential. Intranasal(IN) delivery is a possible strategy to solve problems as IN route could circumvent the brain-blood barrier non-invasively and fit repeated dosage regimens. Herein, we gave an overview of studies and clinical trials involved with IN route and discussed the possibility of employing IN delivery to solve problems in stem cells or EVs-based therapy. We reviewed relevant researches that combining stem cells or EVs-based therapy with IN administration and analyzed benefits brought by IN route. Finally, we proposed possible suggestions to facilitate the development of IN delivery of stem cells or EVs.

关键词： Stem cells extracellular vesicles Central nervous system disorders Intranasal Clinical translation Stroke Neurodegenerative disease Glioma

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Dual-engineered cartilage-targeting extracellular vesicles derived from mesenchymal stem cells enhance osteoarthritis treatment via miR-223/NLRP3/pyroptosis axis:Toward a precision therapy

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Bioactive Materials 2023年第12期30卷 169-183页

作者： Weixuan Liu Anqi Liu Xujun Li Ziyang Sun Zhenghua Sun Yaru Liu Gang Wang Dan Huang Hao Xiong Shiyang Yu Xintao Zhang Cunyi Fan Department of Sports Medicine Peking University Shenzhen HospitalShenzhen Peking University-The Hong Kong University of Science and Technology Medical CenterShenzhen518036China Shanghai Engineering Research Center for Orthopedic Material Innovation and Tissue Regeneration Shanghai201306China Department of Orthopedics Shanghai Sixth People’s Hospital Affiliated to Shanghai Jiao Tong University School of MedicineShanghai200233China Department of Orthodontics Shanghai Stomatological Hospital&School of StomatologyFudan UniversityShanghai200001China Shanghai Key Laboratory of Craniomaxillofacial Development and Diseases Fudan UniversityShanghai200001China Minhang Hospital Fudan UniversityShanghai201199China

Osteoarthritis(OA)is the most common disabling joint disease with no effective disease modifying drugs.extracellular vesicles released by several types of mesenchymal stem cells could promote cartilage repair and ameliorate OA pathology in animal models,representing a novel therapeutic strategy.In this study,we demonstrated that extracellular vesicles derived from human umbilical cord mesenchymal stem cells(hUC-EVs)could maintain chondrocyte homeostasis and alleviate OA,and further revealed a novel molecular mechanism of this therapeutic effect.miR-223,which could directly bind with the 3′UTR of NLRP3 mRNA,was found to be a key miRNA for hUC-EVs to exert beneficial effects on inflammation inhibiting and cartilage protecting.For enhancing the effect on mitigating osteoarthritis,exogenous miR-223 was loaded into hUC-EVs by electroporation,and a collagen II-targeting peptide(WYRGRL)was modified onto the surface of hUC-EVs by genetic engineering to achieve a more targeted and efficient RNA delivery to the cartilage.The dual-engineered EVs showed a maximal effect on inhibiting the NLRP3 inflammasome activation and chondrocyte pyroptosis,and offered excellent results for the treatment of OA.This study provides a novel theoretical basis and a promising therapeutic strategy for the application of engineered extracellular vesicles in OA treatment.

关键词： Osteoarthritis extracellular vesicles NLRP3 inflammasome Pyroptosis Cartilage-targeting

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The role of extracellular vesicles in animal reproduction and diseases

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Journal of Animal Science and Biotechnology 2022年第6期13卷 1634-1656页

作者： Sangiliyandi Gurunathan Min-Hee Kang Hyuk Song Nam Hyung Kim Jin-Hoi Kim Department of Stem Cell and Regenerative Biotechnology Konkuk UniversitySeoul 05029Korea Guangdong Provincial Key Laboratory of Large Animal models for Biomedicine Wuyi UniversityJiangmen 529020China

extracellular vesicles(EVs)are nanosized membrane-enclosed compartments that serve as messengers in cell-to-cell communication,both in normal physiology and in pathological conditions.EVs can transfer functional proteins and genetic information to alter the phenotype and function of recipient cells,which undergo different changes that positively affect their structural and functional integrity.Biological fluids are enriched with several subpopulations of EVs,including exosomes,microvesicles(MVs),and apoptotic bodies carrying several cargoes,such as lipids,proteins,and nucleic acids.EVs associated with the reproductive system are actively involved in the regulation of different physiological events,including gamete maturation,fertilization,and embryo and fetal development.EVs can influence follicle development,oocyte maturation,embryo production,and endometrial-conceptus communication.EVs loaded with cargoes are used to diagnose various diseases,including pregnancy disorders;however,these are dependent on the type of cell of origin and pathological characteristics.EV-derived microRNAs(miRNAs)and proteins in the placenta regulate inflammatory responses and trophoblast invasion through intercellular delivery in the placental microenvironment.This review presents evidence regarding the types of extracellular vesicles,and general aspects of isolation,purification,and characterization of EVs,particularly from various types of embryos.Further,we discuss EVs as mediators and messengers in reproductive biology,the effects of EVs on placentation and pregnancy disorders,the role of EVs in animal reproduction,in the male reproductive system,and mother and embryo cross-communication.In addition,we emphasize the role of microRNAs in embryo implantation and the role of EVs in reproductive and therapeutic medicine.Finally,we discuss the future perspectives of EVs in reproductive biology.

关键词： Embryos extracellular vesicles Implantation MicroRNA Pregnancy disorders

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Neuroprotective effect of mesenchymal stem cellderived extracellular vesicles on optic nerve injury in chronic ocular hypertension

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Neural Regeneration Research 2023年第10期18卷 2301-2306页

作者： Fei Yu Yao Wang Chang-Quan Huang Si-Jie Lin Ru-Xin Gao Ren-Yi Wu Department of Glaucoma Eye Institute and Affiliated Xiamen Eye Center of Xiamen UniversitySchool of MedicineXiamen UniversityXiamenFujian ProvinceChina Fujian Provincial Key Laboratory of Corneal&Ocular Surface Diseases XiamenFujian ProvinceChina Shaanxi Provincial Key Lab of Ophthalmology Shaanxi Clinical Study Center for Ocular DiseaseShaanxi Institute of OphthalmologyFirst Affiliated HospitalMedical SchoolNorthwest UniversityXi’anShaanxi ProvinceChina Eye Institute of Xiamen University Fujian Provincial Key Laboratory of Ophthalmology and Visual ScienceXiamenFujian ProvinceChina Department of Glaucoma Shanghai Peace Eye HospitalShanghaiChina

Mesenchymal stem cells have neuroprotective effects that limit damage to the retina and photoreceptors,and which may be mediated by extracellular vesicles(or exosomes)released by mesenchymal stem cells.To investigate the neuroprotective effect of extracellular vesicles derived from umbilical cord mesenchymal stem cells on glaucoma,we established rat models of chronic ocular hypertension by injecting conjunctival fibroblasts into the anterior chamber to mimic optic nerve injury caused by glaucoma.One week after injury,extracellular vesicles derived from umbilical cord-derived mesenchymal stem cells were injected into the vitreous cavity.We found that extracellular vesicles derived from mesenchymal stem cells substantially reduced retinal damage,increased the number of retinal ganglion cells,and inhibited the activation of caspase-3.These findings suggest that mesenchymal stem cell-derived extracellular vesicles can help alleviate optic nerve injury caused by chronic ocular hypertension,and this effect is achieved by inhibiting cell apoptosis.

关键词： animal model apoptosis chronic glaucoma chronic ocular hypertension extracellular vesicles mesenchymal stem cells neuroprotection rat retinal ganglion cells umbilical cord

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Engineered extracellular vesicles for delivery of siRNA promoting targeted repair of traumatic spinal cord injury

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Bioactive Materials 2023年第5期23卷 328-342页

作者： Yuluo Rong Zhuanghui Wang Pengyu Tang Jiaxing Wang Chengyue Ji Jie Chang Yufeng Zhu Wu Ye Jianling Bai Wei Liu Guoyong Yin Lipeng Yu Xuhui Zhou Weihua Cai Department of Orthopedics The First Affiliated Hospital of Nanjing Medical UniversityNanjing210029JiangsuChina Department of Biostatistics School of Public HealthNanjing Medical UniversityNanjing210029JiangsuChina Department of Orthopedics Second Affiliated Hospital of Naval Medical UniversityShanghai200003China Department of Orthopedics Shanghai General Hospital(Shanghai First People’s Hospital)Shanghai Jiao Tong UniversityShanghai200003China

Spinal cord injury(SCI)is a severe disease of the nervous system that causes irreparable damage and loss of function,for which no effective treatments are available to date.Engineered extracellular vesicles(EVs)carrying therapeutic molecules hold promise as an alternative SCI therapy depending on the specific functionalized EVs and the appropriate engineering strategy.In this study,we demonstrated the design of a drug delivery system of peptide CAQK-modified,siRNA-loaded EVs(C-EVs-siRNA)for SCI-targeted therapy.The peptide CAQK was anchored through a chemical modification to the membranes of EVs isolated from induced neural stem cells(iNSCs).CCL2-siRNA was then loaded into the EVs through electroporation.The modified EVs still maintained the basic properties of EVs and showed favorable targeting and therapeutic effects in vitro and in vivo.C-EVs-siRNA specifically delivered siRNA to the SCI region and was taken up by target cells.C-EVs-siRNA used the inherent anti-inflammatory and neuroreparative functions of iNSCs-derived EVs in synergy with the loaded siRNA,thus enhancing the therapeutic effect against SCI.The combination of targeted modified EVs and siRNA effectively regulated the microenvironmental disturbance after SCI,promoted the transformation of microglia/macrophages from M1 to M2 and limited the negative effects of the inflammatory response and neuronal injury on functional recovery in mice after SCI.Thus,engineered EVs are a potentially feasible and efficacious treatment for SCI,and may also be used to develop targeted treatments for other diseases.

关键词： extracellular vesicles Spinal cord injury siRNA Microglia/macrophages polarization Induced neural stem cells

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