Tspan8-Tumor Extracellular Vesicle-Induced Endothelial Cell and Fibroblast Remodeling Relies on the Target Cell-Selective Response.

Wei Mu,Margot Zöller,Jan Provaznik,Thilo Hackert

doi:10.3390/cells9020319

Wei Mu, Margot Zöller + Show 2 more

Open Access

https://doi.org/10.3390/cells9020319

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Journal: Cells	Publication Date: Jan 29, 2020
Citations: 17	License type: CC BY 4.0

Affiliation: Shanghai Jiao Tong University, Heidelberg University

Abstract

Tumor cell-derived extracellular vesicles (TEX) expressing tetraspanin Tspan8-alpha4/beta1 support angiogenesis. Tspan8-alpha6/beta4 facilitates lung premetastatic niche establishment. TEX-promoted target reprogramming is still being disputed, we explored rat endothelial cell (EC) and lung fibroblast (Fb) mRNA and miRNA profile changes after coculture with TEX. TEX were derived from non-metastatic BSp73AS (AS) or metastatic BSp73ASML (ASML) rat tumor lines transfected with Tspan8 (AS-Tspan8) or Tspan8-shRNA (ASML-Tspan8kd). mRNA was analyzed by deep sequencing and miRNA by array analysis of EC and Fb before and after coculture with TEX. EC and Fb responded more vigorously to AS-Tspan8- than AS-TEX. Though EC and Fb responses differed, both cell lines predominantly responded to membrane receptor activation with upregulation and activation of signaling molecules and transcription factors. Minor TEX-initiated changes in the miRNA profile relied, at least partly, on long noncoding RNA (lncRNA) that also affected chromosome organization and mRNA processing. These analyses uncovered three important points. TEX activate target cell autonomous programs. Responses are initiated by TEX targeting units and are target cell-specific. The strong TEX-promoted lncRNA impact reflects lncRNA shuttling and location-dependent distinct activities. These informations urge for an in depth exploration on the mode of TEX-initiated target cell-specific remodeling including, as a major factor, lncRNA.

Highlights

Extracellular microvesicles (EV) are of utmost importance in intercellular crosstalk in health and disease [1,2,3]
We selected for Fb that contribute to premetastatic niche generation [9,92] and endothelial cells (EC) responding to tumor cell-derived small extracellular vesicles (TEX) after 1h to 5d [50], which we considered a hint for a target cell autonomous response
Few miRNA were recovered at a higher level in TEX than cells, which fits to a selective transfer of miRNA into intraluminal vesicles (ILV)/multivesicular bodies (MVB), where GW182 bodies, possibly in association with miRNA-loaded AGO2 and the RNA-induced silencing complex (RISC), are sorted into MVB for secretion and/or lysosomal degradation [19,94]

Summary

Introduction

Extracellular microvesicles (EV) are of utmost importance in intercellular crosstalk in health and disease [1,2,3]. The mode of action is still disputed, whether the transferred EV content acts autonomously or activates host cell-inherent programs [4,5]. We approached this question analyzing the impact of tumor cell-derived small extracellular vesicles (TEX) on endothelial cells (EC) and fibroblasts (Fb). Exosome lipids show higher lipid order and stability against detergents and are enriched in sphingomyelin, cholesterol, GM3/GRM61 , and PS1 , high level PS expression allowing differentiation from microvesicles [13]

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