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

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.

The Context-Dependent Impact of Integrin-Associated CD151 and Other Tetraspanins on Cancer Development and Progression: A Class of Versatile Mediators of Cellular Function and Signaling, Tumorigenesis and Metastasis

“…Exosomes are highly enriched in tetraspanins, particularly CD63 and CD151, in addition to diverse intracellular components, including diverse RNA species, metabolites, and proteases [ 10 , 121 , 123 ]. Given the nature and signaling capabilities of TEMs, it is of no surprise that many tetraspanins are regarded as key contributors to a variety of exosome-associated functional roles, ranging from tumor cell migration, angiogenesis and signal transduction to expression of critical cancer genes and tumor metabolism through control of micro-RNA or non-coding RNA or metabolite pools [ 67 , 124 , 125 ]. In case of CD151, it is suggested to act in concert with TSPAN8 to drive exosome production in both tumor and endothelial cells, thereby facilitating tumor metastasis [ 30 , 118 ].…”

The Cellular and Biological Impact of Extracellular Vesicles in Pancreatic Cancer

“…It was hypothesized that T-span8 interacts with integrins for MMP transcription or with G-coupled protein receptors (GPCRs), regulating various signaling pathways. In subsequent studies, overexpression of Tspan8 was found to selectively recruit CD49d into EVs, which is essential for their uptake and EC activation in a VEGF-independent manner, specifically promoting proliferation and migration as well as survival and differentiation of EC progenitors [ 83 , 84 ]. Almost a decade later, more simplified in vitro incubation of pancreatic tumor cell-derived EVs with human umbilical vein endothelial cells (HUVEC) led to observations of dynamin-dependent endocytosis of tumor-EVs and enhanced tube formation associated with the activation of Akt and ERK1/2 in ECs [ 85 ].…”

Migrasome and Tetraspanins in Vascular Homeostasis: Concept, Present, and Future

“…Gesierich et al (2006) firstly reported that TSPAN8 is the strongest angiogenesis inducer, as that overexpression of TSPAN8 in tumor cells markedly increases angiogenesis in vivo and in vitro, and co-culture of TSPAN8 knockdown tumor cells or the exosomes-depleted supernatant with HUVECs markedly inhibit HUVECs tube formation in vitro (Gesierich et al, 2006;Akiel et al, 2016). Mechanistically, the tumor cells released exosomes containing TSPAN8 are taken up by target cells via ligands for TSPAN8-associated molecules, and induce angiogenic gene transcription and modulate the RNA profile in ECs or adjacent fibroblasts, and exosomes expressing TSPAN8-CD49d complex preferentially bind ECs, thus initiating an angiogenic loop by inducing TSPAN8 itself expression on sprouting ECs (Gesierich et al, 2006;Nazarenko et al, 2010;Mu et al, 2020). The contribution of TSPAN8 and TSPAN24 on angiogenesis has also been confirmed by TSPAN8-KO mice, TSPAN24-KO mice and TSPAN8/24 double-KO mice (Takeda et al, 2007;Zhao et al, 2018a,b) (Supplementary Table 1).…”

“…The contribution of TSPAN8 and TSPAN24 on angiogenesis has also been confirmed by TSPAN8-KO mice, TSPAN24-KO mice and TSPAN8/24 double-KO mice (Takeda et al, 2007;Zhao et al, 2018a,b) (Supplementary Table 1). Mechanistically, promotion of angiogenesis by tumor-derived exosomes and rescue of impaired angiogenesis in KO mice by wild type-serum exosomes depend on the association of TSPAN8 and TSPAN24 with GPCR and RTK in ECs and tumor cells (Zhao et al, 2018b;Mu et al, 2020). Most importantly, the TSPAN24integrin complex as a functional unit, and the YRSL motif of TSPAN24 plays key role in TSPAN24-mediated angiogenesis (Sincock et al, 1999;Zhang et al, 2002;Zuo et al, 2010;Liu et al, 2011;Peng et al, 2013;Huang et al, 2016).…”

“…The exosomes play essential roles in ECs dysfunction and regeneration, VSMCs migration, ischemic heart disease, cardiac hypertrophy and fibrosis by transferring their bioactive cargos, such as microRNAs and proteins (Ibrahim and Marban, 2016;Barile et al, 2017;Chang et al, 2018;Tong et al, 2018;Baruah and Wary, 2019;Ranjan et al, 2019). Moreover, cancer cells released exosomes containing TSPAN8 are critical for angiogenesis, as that they can be taken up and initiate angiogenic genes transcription and modulate the RNA profile in ECs or adjacent fibroblasts (Gesierich et al, 2006;Nazarenko et al, 2010;Mu et al, 2020). However, it is not known whether the tumor cellsexpressed migraomses can be taken up by ECs in vivo, and influence tumor-induced angiogenesis and subsequently tumor growth, and what will happen if these tumor-derived migrasomes reaches organs distant from the tumor?…”