title: In vivo cleaved CDCP1 promotes early tumor dissemination via complexing with activated β1 integrin and induction of FAK/PI3K/Akt motility signaling.
abstract: Specific cleavage of the transmembrane molecule, CUB domain-containing protein-1 (CDCP1), by plasmin-like serine proteases induces outside-in signal transduction that facilitates early stages of spontaneous metastasis leading to tumor cell intravasation, namely cell escape from the primary tumor, stromal invasion and transendothelial migration. We identified active β1 integrin as a biochemical and functional partner of the membrane-retained 70-kDa CDCP1 fragment, newly generated from its full-length 135-kDa precursor though proteolytic cleavage by serine proteases. Both in cell cultures and in live animals, active β1 integrin complexed preferentially with functionally activated, phosphorylated 70-kDa CDCP1. Complexing of β1 integrin the 70-kDa with CDCP1 fragment induced intracellular phosphorylation signaling, involving focal adhesion kinase-1 (FAK) and PI3 kinase (PI3K)-dependent Akt activation. Thus, inhibition of FAK/PI3K activities by specific inhibitors as well as short-hairpin RNA downregulation of β1 integrin significantly reduced FAK/Akt phosphorylation under conditions where CDCP1 was processed by serine proteases, indicating that FAK/PI3K/Akt pathway operates downstream of cleaved CDCP1 complexed with β1 integrin. Furthermore, this complex-dependent signaling correlated positively with high levels of tumor cell intravasation and dissemination. Correspondingly, abrogation in vivo of CDCP1 cleavage either by unique cleavage-blocking monoclonal antibody 10-D7 or by inhibition of proteolytic activity of plasmin-like serine proteases with aprotinin prevented β1 integrin/CDCP1 complexing and downstream FAK/Akt signaling concomitant with significant reduction of stromal invasion and spontaneous metastasis. Therefore, β1 integrin appears to serve as a motility-regulating partner mediating cross-talk between proteolytically cleaved, membrane-retained CDCP1 and members of FAK/PI3K/Akt pathway. This CDCP1 cleavage-induced signaling cascade constitutes a unique mechanism, independent of extracellular matrix remodeling, whereby a proteolytically cleaved CDCP1 regulates in vivo locomotion and metastasis of tumor cells through β1 integrin partnering. Our findings indicate that CDCP1 cleavage, occurring at the apex of a β1 integrin/FAK/PI3K/Akt signaling cascade, may represent a therapeutic target for CDCP1-positive cancers.
authors.affiliation: The Cell Biology Department, The Scripps Research Institute, La Jolla, CA, USA.
authors.forename: B
authors.initials: B
authors.lastname: Casar
authors.affiliation: The Cell Biology Department, The Scripps Research Institute, La Jolla, CA, USA.
authors.forename: I
authors.initials: I
authors.lastname: Rimann
authors.affiliation: Department of Medicine, University of California San Diego, La Jolla, CA, USA.
authors.forename: H
authors.initials: H
authors.lastname: Kato
authors.affiliation: 1] Department of Medicine, University of California San Diego, La Jolla, CA, USA [2] Moores Cancer Center, University of California San Diego, La Jolla, CA, USA.
authors.forename: S J
authors.initials: SJ
authors.lastname: Shattil
authors.affiliation: The Cell Biology Department, The Scripps Research Institute, La Jolla, CA, USA.
authors.forename: J P
authors.initials: JP
authors.lastname: Quigley
authors.affiliation: The Cell Biology Department, The Scripps Research Institute, La Jolla, CA, USA.
authors.forename: E I
authors.initials: EI
authors.lastname: Deryugina
mesh_terms: D000818:Animals; D015703:Antigens, CD / Q000502:physiology*; D000951:Antigens, Neoplasm; D015815:Cell Adhesion Molecules / Q000502:physiology*; D045744:Cell Line, Tumor; D002465:Cell Movement; D002642:Chick Embryo; D042783:Endothelial Cells / Q000473:pathology; D051417:Focal Adhesion Kinase 1 / Q000502:physiology*; D006801:Humans; D019012:Integrin beta1 / Q000502:physiology*; D008297:Male; D051379:Mice; D009361:Neoplasm Invasiveness; D009362:Neoplasm Metastasis; D009363:Neoplasm Proteins / Q000502:physiology*; D019869:Phosphatidylinositol 3-Kinases / Q000502:physiology*; D010766:Phosphorylation; D011471:Prostatic Neoplasms / Q000473:pathology*; D051057:Proto-Oncogene Proteins c-akt / Q000502:physiology*; D057057:Serine Proteases / Q000502:physiology; D015398:Signal Transduction / Q000502:physiology*
chemical_list: D015703:Antigens, CD; D000951:Antigens, Neoplasm; D015815:Cell Adhesion Molecules; D051417:Focal Adhesion Kinase 1; D019012:Integrin beta1; D009363:Neoplasm Proteins; D019869:Phosphatidylinositol 3-Kinases; D051057:Proto-Oncogene Proteins c-akt; D057057:Serine Proteases; C432044:CDCP1 protein, human