Ovarian Cancer-Driven Mesothelial-to-Mesenchymal Transition is Triggered by the Endothelin-1/β-arr1 Axis

Danila Del Rio,Francesca Spadaro,Manuel López-Cabrera,Flavia Ottavi,Ilenia Masi,Laura Rosanò,Anna Bagnato,Pilar Sandoval,Ricardo Sainz De La Cuesta,Valentina Caprara,Raffaele Strippoli

doi:10.3389/fcell.2021.764375

Abstract

Transcoelomic spread of serous ovarian cancer (SOC) results from the cooperative interactions between cancer and host components. Tumor-derived factors might allow the conversion of mesothelial cells (MCs) into tumor-associated MCs, providing a favorable environment for SOC cell dissemination. However, factors and molecular mechanisms involved in this process are largely unexplored. Here we investigated the tumor-related endothelin-1 (ET-1) as an inducer of changes in MCs supporting SOC progression. Here, we report a significant production of ET-1 from MCs associated with the expression of its cognate receptors, ETA and ETB, along with the protein β-arrestin1. ET-1 triggers MC proliferation via β-arrestin1-dependent MAPK and NF-kB pathways and increases the release of cancer-related factors. The ETA/ETB receptor activation supports the genetic reprogramming of mesothelial-to-mesenchymal transition (MMT), with upregulation of mesenchymal markers, as fibronectin, α-SMA, N-cadherin and vimentin, NF-kB-dependent Snail transcriptional activity and downregulation of E-cadherin and ZO-1, allowing to enhanced MC migration and invasion, and SOC transmesothelial migration. These effects are impaired by either blockade of ETAR and ETBR or by β-arrestin1 silencing. Notably, in peritoneal metastases both ETAR and ETBR are co-expressed with MMT markers compared to normal control peritoneum. Collectively, our report shows that the ET-1 axis may contribute to the early stage of SOC progression by modulating MC pro-metastatic behaviour via MMT.

Highlights

The spreading of serous ovarian cancer (SOC) cells through the peritoneal cavity and the metastasis to the omentum and peritoneum is a common characteristic in patients with advanced tumor (Lengyel, 2010; Yeung et al, 2015)
To evaluate whether a paracrine ET-1 secreted by SOC cells can act as a regulator of mesothelial cells (MCs), we tested the effect of conditioned media (CM) from SKOV3 or HEY cells untreated or treated with the ETAR antagonist Ambrisentan (AMB)
Since activation of NF-kB is related to an aggressive phenotype, plays a crucial role in EMT induction in different cell types including MCs, and its target proteins include Snail (Barberà et al, 2004), we evaluated whether ET-1-dependent NF-κB signaling might be involved in mesenchymal transition (MMT) occurrence

Summary

Introduction

The spreading of serous ovarian cancer (SOC) cells through the peritoneal cavity and the metastasis to the omentum and peritoneum is a common characteristic in patients with advanced tumor (Lengyel, 2010; Yeung et al, 2015). Metastatic spread is largely mediated by the ability of spheroids, and individual cells, to adhere to a monolayer of peritoneal mesothelial cells (MCs) resting upon a basement membrane, to induce their retraction and to penetrate the sub-mesothelial interstitial collagen-rich extracellular matrix (ECM), followed by their survival, and tumor nodule formation with surrounding host cells (Burleson et al, 2004; Shield et al, 2009; Mogi et al, 2021) In this context, the invasive behaviour of SOC spheroids depends upon their contractile capacity and involves both integrins and cadherins (Sodek et al, 2009; Iwanicki et al, 2011; Klymenko et al, 2017a; Klymenko et al, 2017b; SacksSuarez et al, 2019; Goyeneche et al, 2020). The presence of N-cadherin cell-cell junctions in ascitic spheroids as well as matrix metalloprotease (MMP)-mediated proteolysis are key determinants in mesothelial clearance and matrix invasion, predicting more metastatic lesions (Klymenko et al, 2017a; Klymenko et al, 2017b)

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