Institute of Genomics and Multiscale Biology, Icahn School of Medicine at Mount Sinai, New York, NY, 10029, USA.
Department of Genetics and Genomic Sciences, Icahn School of Medicine at Mount Sinai, New York, NY, 10029, USA.
BMC Genomics. 2017 Dec 22;18(1):987. doi: 10.1186/s12864-017-4359-1.
Exosomes and other extracellular vesicles (EVs) have emerged as an important mechanism of cell-to-cell communication. However, previous studies either did not fully resolve what genetic materials were shuttled by exosomes or only focused on a specific set of miRNAs and mRNAs. A more systematic method is required to identify the genetic materials that are potentially transferred during cell-to-cell communication through EVs in an unbiased manner.
In this work, we present a novel next generation of sequencing (NGS) based approach to identify EV mediated mRNA exchanges between co-cultured adipocyte and macrophage cells. We performed molecular and genomic profiling and jointly considered data from RNA sequencing (RNA-seq) and genotyping to track the "sequence varying mRNAs" transferred between cells. We identified 8 mRNAs being transferred from macrophages to adipocytes and 21 mRNAs being transferred in the opposite direction. These mRNAs represented biological functions including extracellular matrix, cell adhesion, glycoprotein, and signal peptides.
Our study sheds new light on EV mediated RNA communications between adipocyte and macrophage cells, which may play a significant role in developing insulin resistance in diabetic patients. This work establishes a new method that is applicable to examining genetic material exchanges in many cellular systems and has the potential to be extended to in vivo studies as well.
外泌体和其他细胞外囊泡(EVs)已成为细胞间通讯的重要机制。然而,以前的研究要么没有完全解决外泌体转运的遗传物质是什么,要么只关注特定的一组 miRNAs 和 mRNAs。需要一种更系统的方法,以无偏倚的方式识别通过 EV 在细胞间通讯过程中潜在转移的遗传物质。
在这项工作中,我们提出了一种新的基于下一代测序(NGS)的方法,用于鉴定共培养的脂肪细胞和巨噬细胞之间 EV 介导的 mRNA 交换。我们进行了分子和基因组分析,并共同考虑了来自 RNA 测序(RNA-seq)和基因分型的数据,以跟踪细胞间转移的“序列变异 mRNAs”。我们鉴定了 8 个从巨噬细胞转移到脂肪细胞的 mRNAs 和 21 个在相反方向转移的 mRNAs。这些 mRNAs 代表了生物学功能,包括细胞外基质、细胞黏附、糖蛋白和信号肽。
我们的研究揭示了脂肪细胞和巨噬细胞之间 EV 介导的 RNA 通讯,这可能在糖尿病患者胰岛素抵抗的发展中起重要作用。这项工作建立了一种新的方法,适用于研究许多细胞系统中的遗传物质交换,并有可能扩展到体内研究。