title: Co-targeting of EGF receptor and neuropilin-1 overcomes cetuximab resistance in pancreatic ductal adenocarcinoma with integrin β1-driven Src-Akt bypass signaling.
abstract: Pancreatic ductal adenocarcinoma (PDAC) cells usually overexpress the epidermal growth factor receptor (EGFR); however, most are resistant to the anti-EGFR monoclonal antibody, cetuximab. In this study, we report that the molecular mechanism of resistance to cetuximab in PDAC cells is mediated by the overexpression of active integrin β1 with downstream Src-Akt activation; this triggers an EGFR ligand-independent proliferation signaling, bypassing EGFR-blocking effect. Knockdown of integrin β1 or inhibition of Src or Akt sensitized cetuximab-resistant (CtxR) PDAC cells to cetuximab. We found that neuropilin-1 (NRP1) physically interacts with active integrin β1, but not inactive one, on the cell surface. To inhibit active integrin β1-driven signaling by targeting NRP1, while suppressing EGFR signaling, we generated an EGFR and NRP1 dual targeting antibody, Ctx-TPP11, by genetic fusion of the NRP1-targeting peptide, TPP11, to the C terminus of the cetuximab heavy chain (Ctx-TPP11). We demonstrate that Ctx-TPP11 efficiently inhibited the growth of CtxRPDAC cells, in vitro and in vivo. The sensitization mechanism involved downregulating active integrin β1 levels through NRP1-coupled internalization mediated by the TPP11 moiety, leading to the inhibition of active integrin β1-driven bypass signaling. Our findings identify aberrant active integrin β1-driven Src-Akt hyperactivation as a primary resistance mechanism to cetuximab in PDAC cells and offer an effective therapeutic strategy to overcome this resistance using an EGFR and NRP1 dual targeting antibody.
authors: Department of Molecular Science and Technology, Ajou University, Suwon, Korea.
authors: Y-J
authors: YJ
authors: Kim
authors: Department of Molecular Science and Technology, Ajou University, Suwon, Korea.
authors: K
authors: K
authors: Jung
authors: Department of Molecular Science and Technology, Ajou University, Suwon, Korea.
authors: D-S
authors: DS
authors: Baek
authors: Department of Biomedical Sciences, College of Medicine, Inha University, Incheon, Korea.
authors: S-S
authors: SS
authors: Hong
authors: Department of Molecular Science and Technology, Ajou University, Suwon, Korea.
authors: Y-S
authors: YS
authors: Kim
mesh_terms: D000230:Adenocarcinoma / Q000188:drug therapy* / Q000235:genetics / Q000473:pathology; D000818:Animals; D021441:Carcinoma, Pancreatic Ductal / Q000188:drug therapy* / Q000235:genetics / Q000473:pathology; D045744:Cell Line, Tumor; D000068818:Cetuximab / Q000008:administration & dosage; D019008:Drug Resistance, Neoplasm / Q000235:genetics*; D066246:ErbB Receptors / Q000037:antagonists & inhibitors / Q000235:genetics*; D006801:Humans; D019012:Integrin beta1 / Q000235:genetics; D051379:Mice; D039942:Neuropilin-1 / Q000037:antagonists & inhibitors / Q000235:genetics*; D015688:Oncogene Protein pp60(v-src) / Q000235:genetics; D051058:Oncogene Protein v-akt / Q000235:genetics; D015398:Signal Transduction / Q000187:drug effects; D023041:Xenograft Model Antitumor Assays
chemical_list: D019012:Integrin beta1; D039942:Neuropilin-1; C512478:EGFR protein, human; D066246:ErbB Receptors; D015688:Oncogene Protein pp60(v-src); D051058:Oncogene Protein v-akt; D000068818:Cetuximab