miR-582-5p Is a Tumor Suppressor microRNA Targeting the Hippo-YAP/TAZ Signaling Pathway in Non-Small Cell Lung Cancer

SIMPLE SUMMARY: Lung cancers lead cancer-related mortalities, with Non-Small Cell Lung Cancer (NSCLC) representing a substantial proportion of these cases. Perturbation of Hippo-YAP/TAZ signaling in NSCLC could be mainly attributed to post-transcriptional regulators since genetic alterations to the signaling pathway are known to be rare. In this study, we identified miR-582-5p as a novel, post-transcriptional regulator of Hippo-YAP/TAZ signaling. Our work revealed an inhibitory function of miR-582-5p on YAP/TAZ signaling in NSCLC cells, whereby the tumorigenic potential is diminished upon the overexpression of miR-582-5p. We also uncovered the regulation of miR-582-5p expression by YAP/TAZ, suggesting a potential feedback loop of YAP/TAZ signaling mediated by miR-582-5p. Mechanistically, we discovered NCKAP1 and PIP5K1C, regulators of actin polymerization, as novel and direct targets of miR-582-5p. Restoring miR-582-5p expression in NSCLC cells resulted in deficient F-actin, which potentially mediates the miR-582-5p-driven tumor suppression in a YAP/TAZ-dependent manner. Our findings underscore the anti-tumor function of miR-582-5p in NSCLC, positing its therapeutic potential in YAP/TAZ-driven lung cancers. ABSTRACT: The Hippo-YAP/TAZ signaling pathway is an evolutionarily conserved signaling pathway involved in a broad spectrum of biological processes, including tumorigenesis. Whilst aberrant Hippo-YAP/TAZ signaling is frequently reported in various cancers, the genetic alterations of this pathway are relatively rare, suggesting regulation at the post-transcriptional level. MicroRNAs play key role in tumorigenesis by regulating gene expression post-transcriptionally. Amongst the cancer-relevant microRNAs, miR-582-5p suppresses cell growth and tumorigenesis by inhibiting the expression of oncogenes, including AKT3, MAP3K2 and NOTCH1. Given the oncogenic role of YAP/TAZ in solid tumors, we scrutinized the possible interplay between miR-582-5p and Hippo-YAP/TAZ signaling. Correlation analysis in NSCLC cells revealed a positive relationship between the expression of mature miR-582-5p and the proportion of phosphorylated YAP/TAZ. Intriguingly, YAP/TAZ knockdown reduced the expression of mature miR-582-5p but increased that of primary miR-582. Overexpression of miR-582-5p resulted in increased phosphorylation of YAP/TAZ with a concomitant reduction in cell proliferation and enhanced apoptosis. Mechanistically, we find that miR-582-5p targets actin regulators NCKAP1 and PIP5K1C, which may be responsible for the observed alteration in F-actin, known to modulate YAP/TAZ. We postulate that regulation of the actin cytoskeleton by miR-582-5p may attenuate YAP/TAZ activity. Altogether, this study reveals a novel mechanism of YAP/TAZ regulation by miR-582-5p in a cytoskeleton-dependent manner and suggests a negative feedback loop, highlighting the therapeutic potential of restoring miR-582-5p expression in treating NSCLC.