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A next generation sequencing based approach to identify extracellular vesicle mediated mRNA transfers between cells

ABSTRACT: Purpose: Exosomes and other extracellular vesicles (EVs) have emerged as an important mechanism of cell-to-cell communication. 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. Methods: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. Results: 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. Conclusions: 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 widely applicable to examining genetic material exchanges in many other cellular systems.

ORGANISM(S): Homo sapiens

PROVIDER: GSE94155 | GEO | 2018/02/13

REPOSITORIES: GEO

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Project description:Cell-free RNAs in biofluids provide opportunities to monitor cancer in a non-invasive manner. Although extracellular microRNAs are extensively characterized, fragmented cell-free long RNAs are not well investigated. Here, we developed Detector-seq (depletion-assisted multiplexing cell-free total RNA sequencing) to enable the deciphering of the cell-free transcriptome. After demonstrating the superior performance of detecting fragmented cell-free long RNAs, we applied Detector-seq to compare cell-free RNAs in human plasma and its extracellular vesicle (EV). Distinct human and microbial RNA signatures were revealed. Structured circular RNA, tRNA, and Y RNA were enriched in plasma, while mRNA and srpRNA were enriched in EV. Meanwhile, cell-free RNAs derived from the virus were more enriched in plasma than in EV. We identified RNAs that showed a selective distribution between plasma and EV and uncovered their distinct functional pathways, that is RNA splicing, antimicrobial humoral response enriched in plasma and transcriptional activity, cell migration, and antigen receptor-mediated immune signals enriched in EV. Although distinctive cancer-relevant RNA signals were identified in plasma and EV, a comparable performance of distinguishing cancer patients from normal individuals could be achieved. Compared to human RNAs, microbe-derived RNA features enabled better classification between colorectal and lung cancer. And for these microbial RNAs, plasma RNAs outperformed EV RNAs for the discrimination of cancer types. Overall, our work provides insights into the unexplored difference of cell-free RNA signals between plasma and EV, thus offering practical guidance for proper selection (with/without EV enrichment) when launching an RNA-based liquid biopsy study. Furthermore, with the ability to capture understudied cell-free long RNA fragments, Detector-seq offers new possibilities for transcriptome-wide characterization of cell-free RNAs to facilitate the understanding of extracellular RNA biology and clinical advances of liquid biopsy.

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A next generation sequencing based approach to identify extracellular vesicle mediated mRNA transfers between cells

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