title: Endothelin A receptor drives invadopodia function and cell motility through the β-arrestin/PDZ-RhoGEF pathway in ovarian carcinoma.
abstract: The endothelin-1 (ET-1)/ET A receptor (ETAR) signalling pathway is a well-established driver of epithelial ovarian cancer (EOC) progression. One key process promoted by ET-1 is tumor cell invasion, which requires the scaffolding functions of β-arrestin-1 (β-arr1) downstream of the receptor; however, the potential role of ET-1 in inducing invadopodia, which are crucial for cellular invasion and tumor metastasis, is completely unknown. We describe here that ET-1/ETAR, through β-arr1, activates RhoA and RhoC GTPase and downstream ROCK (Rho-associated coiled coil-forming kinase) kinase activity, promoting actin-based dynamic remodelling and enhanced cell invasion. This is accomplished by the direct interaction of β-arr1 with PDZ-RhoGEF (postsynaptic density protein 95/disc-large/zonula occludens-RhoGEF). Interestingly, ETAR-mediated invasive properties are related to the regulation of invadopodia, as evaluated by colocalization of actin with cortactin, as well as with TKS5 and MT1-MMP (membrane type 1-matrix metalloproteinase) with areas of matrix degradation, and activation of cofilin pathway, which is crucial for regulating invadopodia activity. Depletion of PDZ-RhoGEF, or β-arr1, or RhoC, as well as the treatment with the dual ET-1 receptor antagonist macitentan, significantly impairs invadopodia function, MMP activity and invasion, demonstrating that β-arr1/PDZ-RhoGEF interaction mediates ETAR-driven ROCK-LIMK-cofilin pathway through the control of RhoC activity. In vivo, macitentan is able to inhibit metastatic dissemination and cofilin phosphorylation. Collectively, our data unveil a noncanonical activation of the RhoC/ROCK pathway through the β-arr1/PDZ-RhoGEF complex as a regulator of ETAR-induced motility and metastasis, establishing ET-1 axis as a novel regulator of invadopodia protrusions through the RhoC/ROCK/LIMK/cofilin pathway during the initial steps of EOC invasion.
authors.affiliation: Regina Elena National Cancer Institute Rome, Rome, Italy.
authors.forename: E
authors.initials: E
authors.lastname: Semprucci
authors.affiliation: Regina Elena National Cancer Institute Rome, Rome, Italy.
authors.forename: P
authors.initials: P
authors.lastname: Tocci
authors.affiliation: Regina Elena National Cancer Institute Rome, Rome, Italy.
authors.forename: R
authors.initials: R
authors.lastname: Cianfrocca
authors.affiliation: Regina Elena National Cancer Institute Rome, Rome, Italy.
authors.forename: R
authors.initials: R
authors.lastname: Sestito
authors.affiliation: Regina Elena National Cancer Institute Rome, Rome, Italy.
authors.forename: V
authors.initials: V
authors.lastname: Caprara
authors.affiliation: Regina Elena National Cancer Institute Rome, Rome, Italy.
authors.forename: M
authors.initials: M
authors.lastname: Veglione
authors.affiliation: Regina Elena National Cancer Institute Rome, Rome, Italy.
authors.forename: V Di
authors.initials: VD
authors.lastname: Castro
authors.affiliation: Section of Experimental Immunotherapy, Department of Haematology, Oncology and Molecular Medicine, Istituto Superiore di Sanita', Rome, Italy.
authors.forename: F
authors.initials: F
authors.lastname: Spadaro
authors.affiliation: Gynecologic Oncology Unit, Catholic University of Rome, Rome, Italy.
authors.forename: G
authors.initials: G
authors.lastname: Ferrandina
authors.affiliation: Regina Elena National Cancer Institute Rome, Rome, Italy.
authors.forename: A
authors.initials: A
authors.lastname: Bagnato
authors.affiliation: Regina Elena National Cancer Institute Rome, Rome, Italy.
authors.forename: L
authors.initials: L
authors.lastname: Rosanò
mesh_terms: D051339:Actin Depolymerizing Factors / Q000378:metabolism; D000199:Actins / Q000378:metabolism; D033942:Adaptor Proteins, Vesicular Transport / Q000378:metabolism; D000818:Animals; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000235:genetics / Q000502:physiology*; D051356:Cortactin / Q000378:metabolism; D005260:Female; D006801:Humans; D015151:Immunoblotting; D054461:Lim Kinases / Q000378:metabolism; D053511:Matrix Metalloproteinase 14 / Q000378:metabolism; D008819:Mice, Nude; D010051:Ovarian Neoplasms / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D000069261:Podosomes / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D034622:RNA Interference; D044022:Receptor, Endothelin A / Q000235:genetics / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D064067:Rho Guanine Nucleotide Exchange Factors / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction / Q000235:genetics; D014183:Transplantation, Heterologous; D000071557:beta-Arrestins / Q000235:genetics / Q000378:metabolism*; D020741:rho GTP-Binding Proteins / Q000378:metabolism; D054460:rho-Associated Kinases / Q000378:metabolism; D000075024:rhoC GTP-Binding Protein
chemical_list: C577234:ARHGEF11 protein, human; D051339:Actin Depolymerizing Factors; D000199:Actins; D033942:Adaptor Proteins, Vesicular Transport; D051356:Cortactin; D044022:Receptor, Endothelin A; D064067:Rho Guanine Nucleotide Exchange Factors; C478582:SH3PXD2A protein, human; D000071557:beta-Arrestins; C515892:LIMK1 protein, human; D054461:Lim Kinases; D054460:rho-Associated Kinases; D053511:Matrix Metalloproteinase 14; C066690:RHOC protein, human; D020741:rho GTP-Binding Proteins; D000075024:rhoC GTP-Binding Protein