Neuron-derived extracellular vesicles promote neurogenesis of neural stem cells

doi:10.12307/2024.186

Abstract

Abstract: BACKGROUND: It has been shown that neural stem cells can differentiate into neurons, astrocytes, and oligodendrocytes. Mesenchymal stem cells-derived extracellular vesicles have also been shown to cross the blood-brain barrier to reach sites of central nervous injury and promote neural repair. However, it is not clear whether neuron-derived extracellular vesicles promote the differentiation of neural stem cells in a direction that is beneficial for neurogenesis.
OBJECTIVE: To investigate whether neuron-derived extracellular vesicles facilitate neural stem cell differentiation towards neurogenesis.
METHODS: Neurons and neural stem cells were extracted from neonatal SD rat cerebral cortex by trypsin digestion. Cell supernatants of neurons were collected. Neuron-derived extracellular vesicles were extracted. Neural stem cells cultured for 10 days were co-cultured with neuron-derived extracellular vesicles or PBS for 7 days. Immunoblotting, immunofluorescence, and RT-qPCR were used to detect proteins specifically expressed by neurons, neural stem cells, oligodendrocytes, and astrocytes.
RESULTS AND CONCLUSION: The neural stem cells co-cultured with neuron-derived extracellular vesicles showed high expression of neuron-specific proteins and oligodendrocyte-specific proteins including β3-tubulin, neurofilament 200 and myelin basic protein, and low expression of astrocyte-specific protein glial fibrillary acidic protein. These results suggest that neuron-derived extracellular vesicles can promote the differentiation of neural stem cells into neurons and oligodendrocytes and prevent the differentiation of neural stem cells into astrocytes.

Key words: neuron, neural stem cell, astrocyte, oligodendrocyte, extracellular vesicle, differentiation, stem cell transplantation, nerve injury

CLC Number:

Li Zhen, Sun Xiao, Xie Yongpeng, Rong Wang, Sun Haitao. Neuron-derived extracellular vesicles promote neurogenesis of neural stem cells[J]. Chinese Journal of Tissue Engineering Research, 2024, 28(25): 3994-3999.

Figures/Tables 5

References

[1] REYNOLDS BA, WEISS S. Generation of neurons and astrocytes from isolated cells of the adult mammalian central nervous system. Science. 1992;255(5052):1707-1710.
[2] XU B, YIN M, YANG Y, et al. Transplantation of neural stem progenitor cells from different sources for severe spinal cord injury repair in rat. Bioact Mater. 2022;23:300-313.
[3] ASSINCK P, DUNCAN GJ, HILTON BJ, et al. Cell transplantation therapy for spinal cord injury. Nat Neurosci. 2017;20(5):637-647.
[4] 陈晓丽, 李少川, 陈久智, 等. 神经干细胞治疗脊髓损伤的研究近况[J].中国医药生物技术,2017,12(5):424-428.
[5] 李晓寅, 杨晓青, 陈淑莲,等. 胶原/丝素蛋白支架联合神经干细胞治疗创伤性脊髓损伤[J].中国组织工程研究,2023,27(6):890-896.
[6] LU P, WOODRUFF G, WANG Y, et al. Long-distance axonal growth from human induced pluripotent stem cells after spinal cord injury. Neuron. 2014;83(4):789-796.
[7] LU P, WANG Y, GRAHAM L, et al. Long-distance growth and connectivity of neural stem cells after severe spinal cord injury. Cell. 2012;150(6):1264-1273.
[8] ROSENZWEIG ES, BROCK JH, LU P, et al. Restorative effects of human neural stem cell grafts on the primate spinal cord. Nat Med. 2018;24(4):484-490.
[9] SALEWSKI RP, MITCHELL RA, LI L, et al. Transplantation of Induced Pluripotent Stem Cell-Derived Neural Stem Cells Mediate Functional Recovery Following Thoracic Spinal Cord Injury Through Remyelination of Axons. Stem Cells Transl Med. 2015;4(7):743-754.
[10] O’SHEA TM, BURDA JE, SOFRONIEW MV. Cell biology of spinal cord injury and repair. J Clin Invest. 2017;127(9):3259-3270.
[11] FITCH MT, SILVER J. CNS injury, glial scars, and inflammation: Inhibitory extracellular matrices and regeneration failure. Exp Neurol. 2008;209(2):294-301.
[12] MATHIEU M, MARTIN-JAULAR L, LAVIEU G, et al. Specificities of secretion and uptake of exosomes and other extracellular vesicles for cell-to-cell communication. Nat Cell Biol. 2019;21(1):9-17.
[13] ZOMER A, MAYNARD C, VERWEIJ FJ, et al. In Vivo imaging reveals extracellular vesicle-mediated phenocopying of metastatic behavior. Cell. 2015;161(5): 1046-1057.
[14] VALADI H, EKSTRÖM K, BOSSIOS A, et al. Exosome-mediated transfer of mRNAs and microRNAs is a novel mechanism of genetic exchange between cells. Nat Cell Biol. 2007;9(6):654-659.
[15] HUANG CC, NARAYANAN R, ALAPATI S, et al. Exosomes as biomimetic tools for stem cell differentiation: Applications in dental pulp tissue regeneration. Biomaterials. 2016;111:103-115.
[16] SHARMA P, MESCI P, CARROMEU C, et al. Exosomes regulate neurogenesis and circuit assembly. Proc Natl Acad Sci U S A. 2019;116(32):16086-16094.
[17] CHOU CH, FAN HC, HUENG DY. Potential of Neural Stem Cell-Based Therapy for Parkinson’s Disease. Parkinsons Dis. 2015;2015:571475.
[18] PARMAR M. Towards stem cell based therapies for Parkinson’s disease. Development. 2018;145(1):dev156117.
[19] HAYASHI Y, LIN HT, LEE CC, et al. Effects of neural stem cell transplantation in Alzheimer’s disease models. J Biomed Sci. 2020;27(1):29.
[20] IM W, LEE ST, CHU K, et al. Stem Cells Transplantation and Huntington’s Disease. Int J Stem Cells. 2009;2(2):102-108.
[21] CHOI KA, CHOI Y, HONG S. Stem cell transplantation for Huntington’s diseases. Methods. 2018;133:104-112.
[22] ZHANG HA, YUAN CX, LIU KF, et al. Neural stem cell transplantation alleviates functional cognitive deficits in a mouse model of tauopathy. Neural Regen Res. 2022;17(1):152-162.
[23] GHOBRIAL GM, ANDERSON KD, DIDIDZE M, et al. Human Neural Stem Cell Transplantation in Chronic Cervical Spinal Cord Injury: Functional Outcomes at 12 Months in a Phase II Clinical Trial. Neurosurgery. 2017;64(CN_suppl_1):87-91.
[24] CURTIS E, MARTIN JR, GABEL B, et al. A First-in-Human, Phase I Study of Neural Stem Cell Transplantation for Chronic Spinal Cord Injury. Cell Stem Cell. 2018; 22(6):941-950.
[25] LEVI AD, OKONKWO DO, PARK P, et al. Emerging Safety of Intramedullary Transplantation of Human Neural Stem Cells in Chronic Cervical and Thoracic Spinal Cord Injury. Neurosurgery. 2018;82(4):562-575.
[26] LEVI AD, ANDERSON KD, OKONKWO DO, et al. Clinical Outcomes from a Multi-Center Study of Human Neural Stem Cell Transplantation in Chronic Cervical Spinal Cord Injury. J Neurotrauma. 2019;36(6):891-902.
[27] SHIN JC, KIM KN, YOO J, et al. Clinical Trial of Human Fetal Brain-Derived Neural Stem/Progenitor Cell Transplantation in Patients with Traumatic Cervical Spinal Cord Injury. Neural Plast. 2015;2015:630932.
[28] WU CC, LIEN CC, HOU WH, et al. Gain of BDNF Function in Engrafted Neural Stem Cells Promotes the Therapeutic Potential for Alzheimer’s Disease. Sci Rep. 2016;6:27358.
[29] MARSH SE, BLURTON-JONES M. Neural stem cell therapy for neurodegenerative disorders: The role of neurotrophic support. Neurochem Int. 2017;106:94-100.
[30] CUMMINGS BJ, UCHIDA N, TAMAKI SJ, et al. Human neural stem cell differentiation following transplantation into spinal cord injured mice: association with recovery of locomotor function. Neurol Res. 2006;28(5):474-481.
[31] HE N, MAO XJ, DING YM, et al. New insights into the biological roles of immune cells in neural stem cells in post-traumatic injury of the central nervous system. Neural Regen Res. 2023;18(9):1908-1916.
[32] HANEY MJ, KLYACHKO NL, ZHAO Y, et al. Exosomes as drug delivery vehicles for Parkinson’s disease therapy. J Control Release. 2015;207:18-30.
[33] GUO S, PERETS N, BETZER O, et al. Intranasal Delivery of Mesenchymal Stem Cell Derived Exosomes Loaded with Phosphatase and Tensin Homolog siRNA Repairs Complete Spinal Cord Injury. ACS Nano. 2019;13(9):10015-10028.
[34] MAROTE A, TEIXEIRA FG, MENDES-PINHEIRO B, et al. MSCs-Derived Exosomes: Cell-Secreted Nanovesicles with Regenerative Potential. Front Pharmacol. 2016;7:231.
[35] PERETS N, HERTZ S, LONDON M, et al. Intranasal administration of exosomes derived from mesenchymal stem cells ameliorates autistic-like behaviors of BTBR mice. Mol Autism. 2018;9:57.
[36] NI H, YANG S, SIAW-DEBRAH F, et al. Exosomes Derived From Bone Mesenchymal Stem Cells Ameliorate Early Inflammatory Responses Following Traumatic Brain Injury. Front Neurosci. 2019;13:14.
[37] WILLIAMS AM, DENNAHY IS, BHATTI UF, et al. Mesenchymal Stem Cell-Derived Exosomes Provide Neuroprotection and Improve Long-Term Neurologic Outcomes in a Swine Model of Traumatic Brain Injury and Hemorrhagic Shock. J Neurotrauma. 2019;36(1):54-60.
[38] REZA-ZALDIVAR EE, HERNÁNDEZ-SAPIÉNS MA, MINJAREZ B, et al. Potential Effects of MSC-Derived Exosomes in Neuroplasticity in Alzheimer’s Disease. Front Cell Neurosci. 2018;12:317.
[39] FÜHRMANN T, ANANDAKUMARAN PN, SHOICHET MS. Combinatorial Therapies After Spinal Cord Injury: How Can Biomaterials Help? Adv Healthc Mater. 2017; 6(10):1601130.
[40] XU B, ZHANG Y, DU XF, et al. Neurons secrete miR-132-containing exosomes to regulate brain vascular integrity. Cell Res. 2017;27(7):882-897.
[41] YIN Z, HAN Z, HU T, et al. Neuron-derived exosomes with high miR-21-5p expression promoted polarization of M1 microglia in culture. Brain Behav Immun. 2020;83:270-282.