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人多能干细胞向心肌细胞分化过程中细胞外囊泡的蛋白质组学分析。

Proteomic Analysis of Exosomes during Cardiogenic Differentiation of Human Pluripotent Stem Cells.

机构信息

Chemical and Biological Engineering, Tufts University, Colby St., Medford, MA 02155, USA.

Cell, Molecular and Developmental Biology, Tufts University, Harrison Ave., Boston, MA 02111, USA.

出版信息

Cells. 2021 Oct 1;10(10):2622. doi: 10.3390/cells10102622.

DOI:10.3390/cells10102622
PMID:34685602
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8533815/
Abstract

Efforts to direct the specification of human pluripotent stem cells (hPSCs) to therapeutically important somatic cell types have focused on identifying proper combinations of soluble cues. Yet, whether exosomes, which mediate intercellular communication, play a role in the differentiation remains unexplored. We took a first step toward addressing this question by subjecting hPSCs to stage-wise specification toward cardiomyocytes (CMs) in scalable stirred-suspension cultures and collecting exosomes. Samples underwent liquid chromatography (LC)/mass spectrometry (MS) and subsequent proteomic analysis revealed over 300 unique proteins from four differentiation stages including proteins such as PPP2CA, AFM, MYH9, MYH10, TRA2B, CTNNA1, EHD1, ACTC1, LDHB, and GPC4, which are linked to cardiogenic commitment. There was a significant correlation of the protein composition of exosomes with the hPSC line and stage of commitment. Differentiating hPSCs treated with exosomes from hPSC-derived CMs displayed improved efficiency of CM formation compared to cells without exogenously added vesicles. Collectively, these results demonstrate that exosomes from hPSCs induced along the CM lineage contain proteins linked to the specification process with modulating effects and open avenues for enhancing the biomanufacturing of stem cell products for cardiac diseases.

摘要

将人类多能干细胞 (hPSC) 定向特化为治疗上重要的体细胞类型的努力集中在鉴定合适的可溶性信号组合上。然而，外泌体是否在分化过程中发挥作用仍未被探索。我们通过在可扩展的搅拌悬浮培养中使 hPSC 分阶段特化为心肌细胞 (CM) 并收集外泌体，从而首次着手解决这个问题。样品经过液相色谱 (LC)/质谱 (MS) 分析和随后的蛋白质组学分析，从四个分化阶段中鉴定出超过 300 种独特蛋白质，包括 PPP2CA、AFM、MYH9、MYH10、TRA2B、CTNNA1、EHD1、ACTG1、LDHB 和 GPC4 等与心脏发生相关的蛋白质。外泌体的蛋白质组成与 hPSC 系和定向分化阶段之间存在显著相关性。与未添加外源性囊泡的细胞相比，用源自 hPSC 的 CM 的外泌体处理的分化 hPSC 显示出 CM 形成效率的提高。总的来说，这些结果表明，沿 CM 谱系诱导的 hPSC 来源的外泌体含有与特异性过程相关的蛋白质，具有调节作用，并为增强用于心脏病的干细胞产品的生物制造开辟了途径。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8969/8533815/ab43f2e10d70/cells-10-02622-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8969/8533815/f51b1ea1cc32/cells-10-02622-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8969/8533815/68569d27bf12/cells-10-02622-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8969/8533815/1b40ad57681c/cells-10-02622-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8969/8533815/4b94d3bf2da0/cells-10-02622-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8969/8533815/ef2fdd2ef297/cells-10-02622-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8969/8533815/0d0cdd054977/cells-10-02622-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8969/8533815/ab43f2e10d70/cells-10-02622-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8969/8533815/f51b1ea1cc32/cells-10-02622-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8969/8533815/68569d27bf12/cells-10-02622-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8969/8533815/1b40ad57681c/cells-10-02622-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8969/8533815/4b94d3bf2da0/cells-10-02622-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8969/8533815/ef2fdd2ef297/cells-10-02622-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8969/8533815/0d0cdd054977/cells-10-02622-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8969/8533815/ab43f2e10d70/cells-10-02622-g007.jpg

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