title: ADAM17 Promotes Motility, Invasion, and Sprouting of Lymphatic Endothelial Cells.
abstract: Tumor-associated lymphatic vessels actively participate in tumor progression and dissemination. ADAM17, a sheddase for numerous growth factors, cytokines, receptors, and cell adhesion molecules, is believed to promote tumor development, facilitating both tumor cell proliferation and migration, as well as tumor angiogenesis. In this work we addressed the issue of whether ADAM17 may also promote tumor lymphangiogenesis. First, we found that ADAM17 is important for the migratory potential of immortalized human dermal lymphatic endothelial cells (LEC). When ADAM17 was stably silenced in LEC, their proliferation was not affected, but: (i) single-cell motility, (ii) cell migration through a 3D Matrigel/collagen type I matrix, and (iii) their ability to form sprouts in a 3D matrix were significantly diminished. The differences in the cell motility between ADAM17-proficient and ADAM17-silenced cells were eliminated by inhibitors of EGFR and HER2, indicating that ADAM17-mediated shedding of growth factors accounts for LEC migratory potential. Interestingly, ADAM17 depletion affected the integrin surface expression/functionality in LEC. ADAM17-silenced cells adhered to plastic, type I collagen, and fibronectin faster than their ADAM17-proficient counterparts. The difference in adhesion to fibronectin was abolished by a cyclic RGD peptide, emphasizing the involvement of integrins in the process. Using a soluble receptor array, we identified BIG-H3 among several candidate proteins involved in the phenotypic and behavioral changes of LEC upon ADAM17 silencing. In additional assays, we confirmed the increased expression of BIG-H3, as well as TGFβ2 in ADAM17-silenced LEC. The antilymphangiogenic effects of ADAM17 silencing in lymphatic endothelial cells suggest further relevance of ADAM17 as a potential target in cancer therapy.
authors.affiliation: Department of Cell Biochemistry, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University in Kraków, Kraków, Poland; Institute of Bioengineering and Swiss Institute for Cancer Research (ISREC), School of Life Sciences, École Polytechnique Fédérale de Lausanne (EPFL), Lausanne, Switzerland; Institute for Molecular Engineering, University of Chicago, Chicago, Illinois, United States of America.
authors.forename: Renata
authors.initials: R
authors.lastname: Mężyk-Kopeć
authors.affiliation: Department of Cell Biochemistry, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University in Kraków, Kraków, Poland.
authors.forename: Barbara
authors.initials: B
authors.lastname: Wyroba
authors.affiliation: Department of Cell Biochemistry, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University in Kraków, Kraków, Poland.
authors.forename: Krystyna
authors.initials: K
authors.lastname: Stalińska
authors.affiliation: Department of Cell Biochemistry, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University in Kraków, Kraków, Poland.
authors.forename: Tomasz
authors.initials: T
authors.lastname: Próchnicki
authors.affiliation: Department of Cell Biochemistry, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University in Kraków, Kraków, Poland.
authors.forename: Karolina
authors.initials: K
authors.lastname: Wiatrowska
authors.affiliation: Institute of Bioengineering and Swiss Institute for Cancer Research (ISREC), School of Life Sciences, École Polytechnique Fédérale de Lausanne (EPFL), Lausanne, Switzerland; Institute for Molecular Engineering, University of Chicago, Chicago, Illinois, United States of America.
authors.forename: Witold W
authors.initials: WW
authors.lastname: Kilarski
authors.affiliation: Institute of Bioengineering and Swiss Institute for Cancer Research (ISREC), School of Life Sciences, École Polytechnique Fédérale de Lausanne (EPFL), Lausanne, Switzerland; Institute for Molecular Engineering, University of Chicago, Chicago, Illinois, United States of America.
authors.forename: Melody A
authors.initials: MA
authors.lastname: Swartz
authors.affiliation: Department of Cell Biochemistry, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University in Kraków, Kraków, Poland.
authors.forename: Joanna
authors.initials: J
authors.lastname: Bereta
mesh_terms: D051722:ADAM Proteins / Q000378:metabolism*; D000072198:ADAM17 Protein; D002448:Cell Adhesion; D002460:Cell Line; D002465:Cell Movement; D049109:Cell Proliferation; D002470:Cell Survival / Q000187:drug effects; D003470:Culture Media; D042783:Endothelial Cells / Q000166:cytology* / Q000378:metabolism*; D066246:ErbB Receptors / Q000378:metabolism; D016326:Extracellular Matrix Proteins / Q000378:metabolism; D020868:Gene Silencing; D006801:Humans; D016023:Integrins / Q000378:metabolism; D042583:Lymphangiogenesis; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D011956:Receptors, Cell Surface / Q000378:metabolism; D015398:Signal Transduction; D016212:Transforming Growth Factor beta / Q000378:metabolism; D053781:Transforming Growth Factor beta2 / Q000378:metabolism; D000099222:betaIG-H3 Protein
chemical_list: D051722:ADAM Proteins; D000072198:ADAM17 Protein; D003470:Culture Media; D066246:ErbB Receptors; D016326:Extracellular Matrix Proteins; D016023:Integrins; D047428:Protein Kinase Inhibitors; D011956:Receptors, Cell Surface; D016212:Transforming Growth Factor beta; D053781:Transforming Growth Factor beta2; D000099222:betaIG-H3 Protein; C000606462:ADAM17 protein, human