Circulating extracellular vesicles from individuals at high-risk of lung cancer induce pro-tumorigenic conversion of stromal cells through transfer of miR-126 and miR-320

Abstract

Background Extracellular vesicles (EVs) containing specific subsets of functional biomolecules are released by all cell types and analysis of circulating EVs can provide diagnostic and prognostic information. To date, little is known regarding the role of EVs both as biomarkers and potential key players in human lung cancer. Methods Plasma EVs were isolated from 40 cancer-free heavy-smokers classified according to a validated 24-microRNA signature classifier (MSC) at high (MSCpos-EVs) or low (MSCneg-EVs) risk to develop lung cancer. EVs origin and functional properties were investigated using in vitro 3D cultures and in vivo models. The prognostic value of miRNAs inside EVs was assessed in training and in validation cohorts of 54 and 48 lung cancer patients, respectively. Results Different membrane composition, biological cargo and pro-tumorigenic activity were observed in MSCpos vs MSCneg-EVs. Mechanistically, in vitro and in vivo results showed that miR-126 and miR-320 from MSCpos-EVs increased pro-angiogenic phenotype of endothelial cells and M2 polarization of macrophage, respectively. MSCpos-EVs prompted 3D proliferation of non-tumorigenic epithelial cells through c-Myc transfer. Moreover, hypoxia was shown to stimulate the secretion of EVs containing c-Myc from fibroblasts, miR-126-EVs from endothelial cells and miR-320-EVs from granulocytes. Lung cancer patients with higher levels of mir-320 into EVs displayed a significantly shorter overall survival in training [HR2.96] and validation sets [HR2.68]. Conclusion Overall our data provide a new perspective on the pro-tumorigenic role of circulating EVs in high risk smokers and highlight the significance of miR-320-EVs as a new prognostic biomarker in lung cancer patients.

Immunomodulatory role of EV-derived non-coding RNA in lung cancer

“…Although the link between miR-27a-3p and the ICOS gene was not directly demonstrated, in vitro experiments showed that EVs from adipocytes silenced for miR-27a-3p displayed higher levels of ICOS + T cells and higher levels of IFN-γ production. Similarly, Peng et al observed a correlation between the up-regulation of EV-miR-125b-5p and T-cell dysfunction at baseline in nonresponsive NSCLC patients undergoing ICI therapy [65] . Moreover, they identified three miRNAs from the miR-320 family (miR-320d, miR-320c, and miR-320b) associated with a poor prognosis and response to ICIs, identifying these miRNAs as potential biomarkers for therapy response [65] .…”

Noncoding RNAs of Extracellular Vesicles in Tumor Angiogenesis: From Biological Functions to Clinical Significance

“…In another study, exosomal miR-23a, miR-1246, and miR-21 could distinguish CRC (all stages) from the control area [128]. Furthermore, higher levels of plasma miR-320-EV and miR-126-EV in patients with highrisk LC can promote angiogenesis and can be significantly associated with poor overall survival [66]. A similar study showed that serum exosomes of patients with GC were rich in lncRNA ZFAS1.…”

“…Pakravan et al showed that the transfer of exosomal miR-100 from human bone marrow MSCs to BC cells downregulated the expression of VEGF in BC-derived cells by regulating the mTOR/HIF-1α signalling axis, thereby inhibiting angiogenesis (Figure 2B) [78]. miR-126 in MSC-derived EVs modulates the pro-angiogenic capacity of ECs by upregulating VEGF [66]. Therefore, fibroblasts, macrophages, and MSCs have been shown to exert synergistic effects to promote tumor angiogenesis in the TME.…”

Increased levels of microRNA‐320 in blood serum and plasma is associated with imminent and advanced lung cancer

“…The expression of candidate miR‐320s was relatively low in both AD and SQ lung tumor tissue and there were no clear differences between tumor and normal lung tissue samples. Elevated miR‐320 expression in plasma extracellular vesicles has been reported to have pro‐tumorigenic activity [ 46 ]. Still, it is not known how blood expression patterns correlate with those in lung tumor tissue, where downregulation of anti‐tumorigenic activity by miR‐320 family members has been observed [ 47 , 48 , 49 ].…”