title: Ectodomain shedding of the cell adhesion molecule Nectin-4 in ovarian cancer is mediated by ADAM10 and ADAM17.
abstract: We previously showed that the cell adhesion molecule Nectin-4 is overexpressed in ovarian cancer tumors, and its cleaved extracellular domain can be detected in the serum of ovarian cancer patients. The ADAM (adisintegrinandmetalloproteinase) proteases are involved in ectodomain cleavage of transmembrane proteins, and ADAM17 is known to cleave Nectin-4 in breast cancer. However, the mechanism of Nectin-4 cleavage in ovarian cancer has not yet been determined. Analysis of ovarian cancer gene microarray data showed that higher expression of Nectin-4, ADAM10, and ADAM17 is associated with significantly decreased progression-free survival. We quantified Nectin-4 shedding from the surface of ovarian cancer cells after stimulation with lysophosphatidic acid. We report that ADAM17 and ADAM10 cleave Nectin-4 and release soluble Nectin-4 (sN4). Small molecule inhibitors and siRNA knockdown of both ADAM proteases confirmed these results. In matched samples from 11 high-grade serous ovarian cancer patients, we detected 2-20-fold more sN4 in ascites fluid than serum. Co-incubation of ovarian cancer cells with ascites fluid significantly increased sN4 shedding, which could be blocked using a dual inhibitor of ADAM10 and ADAM17. Furthermore, we detected RNA for Nectin-4, ADAM10, and ADAM17 in primary ovarian carcinoma tumors, secondary omental metastases, and ascites cells isolated from serous ovarian cancer patients. In a signaling pathway screen, lysophosphatidic acid increased phosphorylation of AKT, EGF receptor, ERK1/2, JNK1/2/3, and c-Jun. Understanding the function of Nectin-4 shedding in ovarian cancer progression is critical to facilitate its development as both a serum biomarker and a therapeutic target for ovarian cancer.
authors.affiliation: From the Departments of Laboratory Medicine and Pathology.
authors.forename: Petra C
authors.initials: PC
authors.lastname: Buchanan
authors.affiliation: From the Departments of Laboratory Medicine and Pathology.
authors.forename: Kristin L M
authors.initials: KLM
authors.lastname: Boylan
authors.affiliation: From the Departments of Laboratory Medicine and Pathology.|Veterinary and Biomedical Sciences, and.
authors.forename: Bruce
authors.initials: B
authors.lastname: Walcheck
authors.affiliation: From the Departments of Laboratory Medicine and Pathology.
authors.forename: Rachel
authors.initials: R
authors.lastname: Heinze
authors.affiliation: Obstetrics, Gynecology, and Women's Health, University of Minnesota, Minneapolis, Minnesota 55455.
authors.forename: Melissa A
authors.initials: MA
authors.lastname: Geller
authors.affiliation: Obstetrics, Gynecology, and Women's Health, University of Minnesota, Minneapolis, Minnesota 55455.
authors.forename: Peter A
authors.initials: PA
authors.lastname: Argenta
authors.affiliation: From the Departments of Laboratory Medicine and Pathology, skubi002@umn.edu.
authors.forename: Amy P N
authors.initials: APN
authors.lastname: Skubitz
mesh_terms: D000072197:ADAM10 Protein / Q000235:genetics / Q000378:metabolism*; D000072198:ADAM17 Protein / Q000235:genetics / Q000378:metabolism*; D053829:Amyloid Precursor Protein Secretases / Q000235:genetics / Q000378:metabolism*; D015815:Cell Adhesion Molecules / Q000235:genetics / Q000378:metabolism*; D045744:Cell Line, Tumor; D048049:Extracellular Signal-Regulated MAP Kinases / Q000235:genetics / Q000378:metabolism; D005260:Female; D006801:Humans; D020935:MAP Kinase Signaling System / Q000235:genetics; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D009362:Neoplasm Metastasis; D010051:Ovarian Neoplasms / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D000072417:Protein Domains; D051057:Proto-Oncogene Proteins c-akt / Q000235:genetics / Q000378:metabolism; D016755:Proto-Oncogene Proteins c-jun / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012334:RNA, Neoplasm / Q000235:genetics / Q000378:metabolism; D000075983:Nectins
chemical_list: D000072197:ADAM10 Protein; D000072198:ADAM17 Protein; D053829:Amyloid Precursor Protein Secretases; D015815:Cell Adhesion Molecules; D048049:Extracellular Signal-Regulated MAP Kinases; D008565:Membrane Proteins; D051057:Proto-Oncogene Proteins c-akt; D016755:Proto-Oncogene Proteins c-jun; D012333:RNA, Messenger; D012334:RNA, Neoplasm; C442228:NECTIN4 protein, human; D000075983:Nectins; C491236:ADAM10 protein, human; C000606462:ADAM17 protein, human
keywords: ADAM; ADAM10; ADAM17; Nectin-4; biomarker; cancer biology; cell migration; ovarian cancer