Antibody library-based tumor endothelial cells surface proteomic functional screen reveals migration-stimulating factor as an anti-angiogenic target.
Angiogenesis is critical for cancer development and metastasis. Here we have employed a functional antibody library-based proteomic screen to identify proteins that participate in and might be used as therapeutic targets for tumor-related angiogenesis. Mice were immunized with human esophageal cancer endothelial cells (HECEC). The antibody library was established with the mouse spleen cells the serum of which had most anti-angiogenic effect. Monoclonal antibodies were subjected to an immunoreactive and functional screen and monoclonal antibodies that reacted strongly with cell surface antigens of HECECs and influenced their behavior were selected. Antigens that recognized by the antibodies were obtained by immunoprecipitation and then identified by mass spectrometry analysis. Migration-stimulating factor (MSF), the antigen of 1D2 antibody was identified using this approach. Further studies demonstrated that the 1D2 antibody suppressed MSF-effected migration and adhesion of HECECs on fibronectin matrix. Biodistribution assay showed that MSF targeting antibody 1D2 could specifically home to the xenograft with humanized blood vessel. Targeting treatment with 1D2 antibody significantly suppressed tumor growth through inhibition of human tumor-related angiogenesis. These results indicate that the functional antibody library-based proteomic screen can successfully identify proteins that involved in tumor-related angiogenesis and MSF may be a target for the anti-angiogenic treatment of the esophageal cancer.
Hu
Hai
H
State Key Laboratory of Molecular Oncology, Cancer Institute (Hospital), Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing 100021, China.
Ran
Yuliang
Y
Zhang
Yushan
Y
Zhou
Zhuan
Z
Harris
Simon J
SJ
Yu
Long
L
Sun
Lixin
L
Pan
Jian
J
Liu
Jun
J
Lou
Jinning
J
Yang
Zhihua
Z
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000912:Antibodies, Neoplasm / Q000276:immunology*; D000918:Antibody Specificity; D000951:Antigens, Neoplasm / Q000276:immunology; D002448:Cell Adhesion; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000276:immunology / Q000473:pathology; D002465:Cell Movement; D049109:Cell Proliferation; D018920:Coculture Techniques; D016207:Cytokines / Q000032:analysis*; D042783:Endothelial Cells / Q000276:immunology / Q000473:pathology*; D004938:Esophageal Neoplasms / Q000098:blood supply* / Q000276:immunology / Q000473:pathology; D005109:Extracellular Matrix / Q000378:metabolism; D005260:Female; D005353:Fibronectins; D006801:Humans; D007114:Immunization; D051379:Mice; D009389:Neovascularization, Pathologic / Q000276:immunology*; D009500:Neutralization Tests; D019151:Peptide Library; D040901:Proteomics / Q000379:methods*; D023041:Xenograft Model Antitumor Assays
D000911:Antibodies, Monoclonal; D000912:Antibodies, Neoplasm; D000951:Antigens, Neoplasm; D016207:Cytokines; C067625:FN1 protein, human; D005353:Fibronectins; D019151:Peptide Library