학술논문

Sex differences in the transcriptome of extracellular vesicles secreted by fetal neural stem cells and effects of chronic alcohol exposure.
Document Type
Article
Source
Biology of Sex Differences. 4/15/2023, Vol. 14 Issue 1, p1-31. 31p.
Subject
*NEURAL stem cells
*DEVELOPMENTAL neurobiology
*EXTRACELLULAR vesicles
*FETAL brain
*TRANSCRIPTOMES
*RIBOSOMAL RNA
*SEX (Biology)
Language
ISSN
2042-6410
Abstract
Background: Prenatal alcohol (ethanol) exposure (PAE) results in brain growth restriction, in part, by reprogramming self-renewal and maturation of fetal neural stem cells (NSCs) during neurogenesis. We recently showed that ethanol resulted in enrichment of both proteins and pro-maturation microRNAs in sub-200-nm-sized extracellular vesicles (EVs) secreted by fetal NSCs. Moreover, EVs secreted by ethanol-exposed NSCs exhibited diminished efficacy in controlling NSC metabolism and maturation. Here we tested the hypothesis that ethanol may also influence the packaging of RNAs into EVs from cell-of-origin NSCs. Methods: Sex-specified fetal murine iso-cortical neuroepithelia from three separate pregnancies were maintained ex vivo, as neurosphere cultures to model the early neurogenic niche. EVs were isolated by ultracentrifugation from NSCs exposed to a dose range of ethanol. RNA from paired EV and cell-of-origin NSC samples was processed for ribosomal RNA-depleted RNA sequencing. Differential expression analysis and exploratory weighted gene co-expression network analysis (WGCNA) identified candidate genes and gene networks that were drivers of alterations to the transcriptome of EVs relative to cells. Results: The RNA content of EVs differed significantly from cell-of-origin NSCs. Biological sex contributed to unique transcriptome variance in EV samples, where > 75% of the most variant transcripts were also sex-variant in EVs but not in cell-of-origin NSCs. WGCNA analysis also identified sex-dependent enrichment of pathways, including dopamine receptor binding and ectoderm formation in female EVs and cell-substrate adhesion in male EVs, with the top significant DEGs from differential analysis of overall individual gene expressions, i.e., Arhgap15, enriched in female EVs, and Cenpa, enriched in male EVs, also serving as WCGNA hub genes of sex-biased EV WGCNA clusters. In addition to the baseline RNA content differences, ethanol exposure resulted in a significant dose-dependent change in transcript expression in both EVs and cell-of-origin NSCs that predominantly altered sex-invariant RNAs. Moreover, at the highest dose, ~ 73% of significantly altered RNAs were enriched in EVs, but depleted in NSCs. Conclusions: The EV transcriptome is distinctly different from, and more sex-variant than, the transcriptome of cell-of-origin NSCs. Ethanol, a common teratogen, results in dose-dependent sorting of RNA transcripts from NSCs to EVs which may reprogram the EV-mediated endocrine environment during neurogenesis. Highlights: The RNA content of EVs differ significantly from their cell-of-origin NSCs. Biological sex contributes to significant differences in the transcriptome of EVs. Most sex differences observed in EVs were not present in parent-of-origin NSCs, suggesting sex-dependent sorting of RNA into EVs. Ethanol exposure resulted in specific RNA transcripts being enriched in secreted EVs, while being depleting in cell-of-origin NSCs, suggesting that cell stress may alter the sorting of RNA from cells to EVs. Transcripts enriched in EVs, following ethanol exposure of NSCs, encode proteins selectively overrepresented in biological pathways like mRNA splicing and transport. [ABSTRACT FROM AUTHOR]