TY - JOUR
T1 - Extracellular vesicles from human iPSC-derived neural stem cells
T2 - miRNA and protein signatures, and anti-inflammatory and neurogenic properties
AU - Upadhya, Raghavendra
AU - Madhu, Leelavathi N.
AU - Attaluri, Sahithi
AU - Gitaí, Daniel Leite Góes
AU - Pinson, Marisa R.
AU - Kodali, Maheedhar
AU - Shetty, Geetha
AU - Zanirati, Gabriele
AU - Kumar, Smrithi
AU - Shuai, Bing
AU - Weintraub, Susan T.
AU - Shetty, Ashok K.
N1 - Funding Information:
This work was supported by grants from the National Institute of Neurological Disorders and Stroke (1R01NS106907-01 to A.K.S) and the State of Texas (Emerging Technology Fund to A.K.S.). DLGG was supportedby the Research Productivity Scholarship Program in Brazilian National Council for Scientific and Technological Development (CNPq). Gabriele Zanirati was supported by a doctoral fellowship from Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), Governmentof Brazil. Mass spectrometry analyses were conducted in the Institutional Mass Spectrometry Laboratory of the University of Texas Health Science Center at San Antonio. The expert technical assistance of Sammy Pardo and Dana Molleur is greatly appreciated. Fund for purchase of the Lumos mass spectrometer were provided by the University of Texas System Proteomics Core Network (to S.T.W.).
Funding Information:
This work was supported by the National Institute of Neurological Disorders and Stroke [1R01NS106907-01]; Texas Emerging Technology Fund [State of Texas]. This work was supported by grants from the National Institute of Neurological Disorders and Stroke (1R01NS106907-01 to A.K.S) and the State of Texas (Emerging Technology Fund to A.K.S.). DLGG was supportedby the Research Productivity Scholarship Program in Brazilian National Council for Scientific and Technological Development (CNPq). Gabriele Zanirati was supported by a doctoral fellowship from Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior (CAPES), Governmentof Brazil. Mass spectrometry analyses were conducted in the Institutional Mass Spectrometry Laboratory of the University of Texas Health Science Center at San Antonio. The expert technical assistance of Sammy Pardo and Dana Molleur is greatly appreciated. Fund for purchase of the Lumos mass spectrometer were provided by the University of Texas System Proteomics Core Network (to S.T.W.).
Publisher Copyright:
© 2020 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group on behalf of The International Society for Extracellular Vesicles.
PY - 2020/1/1
Y1 - 2020/1/1
N2 - Grafting of neural stem cells (NSCs) derived from human induced pluripotent stem cells (hiPSCs) has shown promise for brain repair after injury or disease, but safety issues have hindered their clinical application. Employing nano-sized extracellular vesicles (EVs) derived from hiPSC-NSCs appears to be a safer alternative because they likely have similar neuroreparative properties as NSCs and are amenable for non-invasive administration as an autologous or allogeneic off-the-shelf product. However, reliable methods for isolation, characterization and testing the biological properties of EVs are critically needed for translation. We investigated signatures of miRNAs and proteins and the biological activity of EVs, isolated from hiPSC-NSCs through a combination of anion-exchange chromatography (AEC) and size-exclusion chromatography (SEC). AEC and SEC facilitated the isolation of EVs with intact ultrastructure and expressing CD9, CD63, CD81, ALIX and TSG 101. Small RNA sequencing, proteomic analysis, pathway analysis and validation of select miRNAs and proteins revealed that EVs were enriched with miRNAs and proteins involved in neuroprotective, anti-apoptotic, antioxidant, anti-inflammatory, blood-brain barrier repairing, neurogenic and Aβ reducing activities. Besides, EVs comprised miRNAs and/or proteins capable of promoting synaptogenesis, synaptic plasticity and better cognitive function. Investigations using an in vitro macrophage assay and a mouse model of status epilepticus confirmed the anti-inflammatory activity of EVs. Furthermore, the intranasal administration of EVs resulted in the incorporation of EVs by neurons, microglia and astrocytes in virtually all adult rat and mouse brain regions, and enhancement of hippocampal neurogenesis. Thus, biologically active EVs containing miRNAs and proteins relevant to brain repair could be isolated from hiPSC-NSC cultures, making them a suitable biologic for treating neurodegenerative disorders.
AB - Grafting of neural stem cells (NSCs) derived from human induced pluripotent stem cells (hiPSCs) has shown promise for brain repair after injury or disease, but safety issues have hindered their clinical application. Employing nano-sized extracellular vesicles (EVs) derived from hiPSC-NSCs appears to be a safer alternative because they likely have similar neuroreparative properties as NSCs and are amenable for non-invasive administration as an autologous or allogeneic off-the-shelf product. However, reliable methods for isolation, characterization and testing the biological properties of EVs are critically needed for translation. We investigated signatures of miRNAs and proteins and the biological activity of EVs, isolated from hiPSC-NSCs through a combination of anion-exchange chromatography (AEC) and size-exclusion chromatography (SEC). AEC and SEC facilitated the isolation of EVs with intact ultrastructure and expressing CD9, CD63, CD81, ALIX and TSG 101. Small RNA sequencing, proteomic analysis, pathway analysis and validation of select miRNAs and proteins revealed that EVs were enriched with miRNAs and proteins involved in neuroprotective, anti-apoptotic, antioxidant, anti-inflammatory, blood-brain barrier repairing, neurogenic and Aβ reducing activities. Besides, EVs comprised miRNAs and/or proteins capable of promoting synaptogenesis, synaptic plasticity and better cognitive function. Investigations using an in vitro macrophage assay and a mouse model of status epilepticus confirmed the anti-inflammatory activity of EVs. Furthermore, the intranasal administration of EVs resulted in the incorporation of EVs by neurons, microglia and astrocytes in virtually all adult rat and mouse brain regions, and enhancement of hippocampal neurogenesis. Thus, biologically active EVs containing miRNAs and proteins relevant to brain repair could be isolated from hiPSC-NSC cultures, making them a suitable biologic for treating neurodegenerative disorders.
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U2 - 10.1080/20013078.2020.1809064
DO - 10.1080/20013078.2020.1809064
M3 - Article
AN - SCOPUS:85089994115
SN - 2001-3078
VL - 9
JO - Journal of Extracellular Vesicles
JF - Journal of Extracellular Vesicles
IS - 1
M1 - 1809064
ER -
