A humanized anti-DLL4 antibody promotes dysfunctional angiogenesis and inhibits breast tumor growth.
Blockage of Delta-like 4 (DLL4)-directed Notch signaling induces excessive tip cell formation and endothelial proliferation resulting in dysfunctional angiogenesis in tumors. MMGZ01, as a murine anti-human DLL4 monoclonal antibody, specifically binds to human DLL4 and blocks Notch pathway. Here, the structure of MMGZ01 variable fragment (Fv) was established and framework region (FR) residues which supported complementarily determining region (CDR) loop conformation were identified. Important residues interactions were also identified through docking MMGZ01 Fv with antigen epitope in DLL4. To humanize the murine antibody, we modified MMGZ01 Fv through CDR grafting and the reconstructed antibody (H3L2) maintained similar structure and binding affinity to parental MMGZ01 after back mutation of 12 canonical murine residues in the FRs. Meanwhile, H3L2 promoted human umbilical vein endothelial cell (HUVEC) proliferation through inhibiting DLL4-directed Notch pathway. Moreover, in MDA-MB-231-bearing nude mice, H3L2 induced dysfunctional angiogenesis and tumor cell apoptosis and showed superior anti-tumor activity. In conclusion, H3L2 is an ideal humanized antibody that inhibits tumor growth through targeting DLL4-Notch pathway and has attracting potentials for clinical applications.
State Key Laboratory of Natural Medicines, School of Life Science &Technology, China Pharmaceutical University, Nanjing, Jiangsu 210009, China.
Xuelian
X
Jia
State Key Laboratory of Natural Medicines, School of Life Science &Technology, China Pharmaceutical University, Nanjing, Jiangsu 210009, China.|The Rutgers Center for Computational and Integrative Biology, Rutgers University, Camden, NJ 08102, United States.
Wenyi
W
Wang
State Key Laboratory of Natural Medicines, School of Life Science &Technology, China Pharmaceutical University, Nanjing, Jiangsu 210009, China.
Zhuobin
Z
Xu
State Key Laboratory of Natural Medicines, School of Life Science &Technology, China Pharmaceutical University, Nanjing, Jiangsu 210009, China.
Shijing
S
Wang
State Key Laboratory of Natural Medicines, School of Life Science &Technology, China Pharmaceutical University, Nanjing, Jiangsu 210009, China.
Tong
T
Wang
State Key Laboratory of Natural Medicines, School of Life Science &Technology, China Pharmaceutical University, Nanjing, Jiangsu 210009, China.
Min
M
Wang
State Key Laboratory of Natural Medicines, School of Life Science &Technology, China Pharmaceutical University, Nanjing, Jiangsu 210009, China.
Min
M
Wu
D048868:Adaptor Proteins, Signal Transducing; D000818:Animals; D000911:Antibodies, Monoclonal / Q000008:administration & dosage* / Q000737:chemistry / Q000235:genetics; D000970:Antineoplastic Agents / Q000008:administration & dosage* / Q000737:chemistry; D001943:Breast Neoplasms / Q000188:drug therapy*; D002135:Calcium-Binding Proteins; D004195:Disease Models, Animal; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000276:immunology*; D008819:Mice, Nude; D062105:Molecular Docking Simulation; D009389:Neovascularization, Pathologic; D011487:Protein Conformation; D051880:Receptors, Notch / Q000378:metabolism; D015398:Signal Transduction; D016896:Treatment Outcome
D048868:Adaptor Proteins, Signal Transducing; D000911:Antibodies, Monoclonal; D000970:Antineoplastic Agents; D002135:Calcium-Binding Proteins; C424126:DLL4 protein, human; D036341:Intercellular Signaling Peptides and Proteins; D051880:Receptors, Notch