Retinal progenitor cells release extracellular vesicles containing developmental transcription factors, microRNA and membrane proteins

视网膜祖细胞释放含有发育转录因子、microRNA 和膜蛋白的细胞外囊泡

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作者:Jing Zhou, Alberto Benito-Martin, Jason Mighty, Lynne Chang, Shima Ghoroghi, Hao Wu, Madeline Wong, Sara Guariglia, Petr Baranov, Michael Young, Rajendra Gharbaran, Mark Emerson, Milica Tesic Mark, Henrik Molina, M Valeria Canto-Soler, Hector Peinado Selgas, Stephen Redenti

Abstract

A range of cell types, including embryonic stem cells, neurons and astrocytes have been shown to release extracellular vesicles (EVs) containing molecular cargo. Across cell types, EVs facilitate transfer of mRNA, microRNA and proteins between cells. Here we describe the release kinetics and content of EVs from mouse retinal progenitor cells (mRPCs). Interestingly, mRPC derived EVs contain mRNA, miRNA and proteins associated with multipotency and retinal development. Transcripts enclosed in mRPC EVs, include the transcription factors Pax6, Hes1, and Sox2, a mitotic chromosome stabilizer Ki67, and the neural intermediate filaments Nestin and GFAP. Proteomic analysis of EV content revealed retinogenic growth factors and morphogen proteins. mRPC EVs were shown to transfer GFP mRNA between cell populations. Finally, analysis of EV mediated functional cargo delivery, using the Cre-loxP recombination system, revealed transfer and uptake of Cre+ EVs, which were then internalized by target mRPCs activating responder loxP GFP expression. In summary, the data supports a paradigm of EV genetic material encapsulation and transfer within RPC populations. RPC EV transfer may influence recipient RPC transcriptional and post-transcriptional regulation, representing a novel mechanism of differentiation and fate determination during retinal development.

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