Comparative Analysis of microRNA Expression Profiles of Exosomes Derived from Normal and Hypoxic Preconditioning Human Neural Stem Cells by Next Generation Sequencing

Abstract

Stroke recovery is associated with neural stem cell (NSC) development and neurovascular unit reconstruction. The exosome, as an important intercellular player in neurovascular communication, mediates neuro-restorative events by transferring exosomal protein and RNA cargoes. In this study, we explored the role of exosomal microRNAs (miRNAs) in human NSCs (hNSCs), and analyzed the expression profiles of miRNAs in hNSC-derived and hypoxic preconditioning hNSC-derived exosomes with the help of next generation sequencing (NGS). The results demonstrated that a certain proportion of miRNAs were differentially expressed in both exosomes. In addition, target gene prediction and Gene Ontology (GO) enrichment analysis showed that these genes were associated with differential miRNAs primarily participating in biological processes (regulation of cellular process), cellular component (intracellular membrane-bounded organelle), and molecular function (binding). Kyoto Encyclopedia of Genes and Genomes pathway (KEGG) pathway enrichment data suggested that most of targeted genes involved in PI3K-Akt, Hippo, MAPK, mTOR, and Endocytosis etc. signaling pathways. We identified the interesting and important expressed miRNA and considered that miR-98-3p might be a special hNSC-derived exosomal-miRNA which was significantly downregulated under hypoxic preconditioning. The hNSCs-derived exosomes were capable of modulating gene expression or promoting stroke therapy. We observed that after hypoxic preconditioning, the functions of these exosomes were changed, and exosomal-miRNAs expression profile was different. In summary, our study suggested that hNSC-derived exosomal miRNAs including hypoxic preconditioning exosomal miRNAs provided a new strategy for the diagnosis and treatment of stroke patients.

How hypoxia regulate exosomes in ischemic diseases and cancer microenvironment?

“…Zhang et al indicated a significant downregulation of exosomal miRNA‐98‐3p after hypoxic preconditioning of the NSCs. They suggested hypoxic preconditioning as a promising strategy for diagnosis and therapy of stroke patients . Hypoxia promoted the production and aggregation of exosomal amyloid‐β in Alzheimer's disease (AD) …”

Preconditioning and Cellular Engineering to Increase the Survival of Transplanted Neural Stem Cells for Motor Neuron Disease Therapy

“…After hypoxic preconditioning, the exosomal miRNA expression profile was modified, and some miRNAs were differentially expressed in the two populations, such as miR-98-3p. Exosomal functions changed as well, and exosomes from preconditioned cells appeared capable of modulating gene expression and promoting stroke recovery [54]. Notably, this work reveals how the administration of exosomes derived from preconditioned cells could represent an alternative approach to direct NSC transplantation, thus avoiding the risks related to the use of a cellular product.…”

“…To further elucidate the molecular mechanisms underlying hypoxic preconditioning, a recent study analyzed the microRNA expression profiles of exosomes derived from normal vs hypoxic preconditioned NSCs by next-generation sequencing [54]. After hypoxic preconditioning, the exosomal miRNA expression profile was modified, and some miRNAs were differentially expressed in the two populations, such as miR-98-3p.…”

Extracellular Vesicles: The New Frontier of Stem Cell Regenerative Medicine?

“…Next generation sequencing revealed that EVs originating from hypoxic human neural stem cells have an increased levels of 53 miRNAs and a reduction in 26 miRNAs [205]. Similarly microarray analysis of EV released by cardiac progenitor cells under hypoxic stress condition showed an upregulation of 11 miRNAs compared to EVs secreted from normoxic cells [206].…”