To identify specific protein antigens that functionally contribute to human tumor cell metastasis, our laboratory initiated subtractive immunization approaches to raise unique monoclonal antibodies (mAbs) that inhibit human tumor dissemination. Panels of generated mAbs are screened for their ability to inhibit human tumor dissemination in chick embryo and mouse models of metastasis. Using this approach, we have generated unique antibodies. One of them, mAb 1A5, recognizes a specific member of the tetraspanin family, CD151, and correspondingly inhibits metastasis by >90%. Another subtractive immunization antibody, mAb 41-2, recognizes a tumor cell transmembrane protein, CDCP1/SIMA-135. This mAb 41-2 inhibits overall metastasis by 60-80%, however the mechanism of mAb 41-2 and that of its target antigen are unknown. Thus, a proposed goal of this renewal is to elucidate the metastasis-inhibition mechanism of mAb 41-2 and to conduct structure- function analysis of its cognate antigen, CDCP1, in order to determine the contributory role of CDCP1 in cancer dissemination. Timed additions of mAb 41-2 into animals followed by quantitative measurements of tumor cell intravascular arrest, apoptosis, survival, extravasation and establishment of secondary foci will determine when, where and how mAb 41-2 blocks malignant function. These approaches will be expanded and confirmed in mouse models of tumor dissemination, including orthotopic implantation approaches. That mAb 41-2's antigen, CDCP1, functions as a survival factor will be tested with mutational analyses of CDCP1's cytoplasmic domain and CDCP1's extracellular CUB domains. The use of combinatorial libraries will be employed to identify unknown natural ligand(s) for CDCP1 and to generate new distinctive antibodies to CDCP1. Another aim of this renewal is to characterize additional pairs of congenic human tumor variants that are similar in tumor-forming ability but differ substantially in their metastasis properties, and to use these cell pairs in new subtractive immunization approaches to generate novel function-blocking antibodies. Panels of mAbs will be screened for differential reactivity and function-blocking properties in our established models. We also propose to initiate potential translational approaches and generate new metastasis-blocking, single chain variable fragment (sc Fv) humanized antibodies that are effective in cancer type-specific orthotopic mouse models and could represent unique reagents for possible targeted tumor therapy.

Public Health Relevance

Deaths from cancer occur mainly because tumor cells spread from the primary tumor site to other vital organs and tissues. In order for tumor cells to escape from the primary tumor and travel to distant organs, they produce different levels of functioning proteins. The goal of the proposed research is to use a modified system of monoclonal antibody generation in order to identify the relevant proteins that contribute to tumor cell spread. Oncologists could target those critical protein molecules and might also employ the specific antibodies as therapeutic agents.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA105412-09
Application #
8259158
Study Section
Tumor Progression and Metastasis Study Section (TPM)
Program Officer
Snyderwine, Elizabeth G
Project Start
2004-01-16
Project End
2014-05-31
Budget Start
2012-06-01
Budget End
2013-05-31
Support Year
9
Fiscal Year
2012
Total Cost
$331,909
Indirect Cost
$157,128
Name
Scripps Research Institute
Department
Type
DUNS #
781613492
City
La Jolla
State
CA
Country
United States
Zip Code
92037
Casar, B; Rimann, I; Kato, H et al. (2014) In vivo cleaved CDCP1 promotes early tumor dissemination via complexing with activated *1 integrin and induction of FAK/PI3K/Akt motility signaling. Oncogene 33:255-68
Deryugina, Elena I; Zajac, Ewa; Juncker-Jensen, Anna et al. (2014) Tissue-infiltrating neutrophils constitute the major in vivo source of angiogenesis-inducing MMP-9 in the tumor microenvironment. Neoplasia 16:771-88
Juncker-Jensen, Anna; Deryugina, Elena I; Rimann, Ivo et al. (2013) Tumor MMP-1 activates endothelial PAR1 to facilitate vascular intravasation and metastatic dissemination. Cancer Res 73:4196-211
Low-Marchelli, Janine M; Ardi, Veronica C; Vizcarra, Edward A et al. (2013) Twist1 induces CCL2 and recruits macrophages to promote angiogenesis. Cancer Res 73:662-71
Quigley, James P; Deryugina, Elena I (2012) Combating angiogenesis early: potential of targeting tumor-recruited neutrophils in cancer therapy. Future Oncol 8:5-8
Bekes, Erin M; Deryugina, Elena I; Kupriyanova, Tatyana A et al. (2011) Activation of pro-uPA is critical for initial escape from the primary tumor and hematogenous dissemination of human carcinoma cells. Neoplasia 13:806-21
Bekes, Erin M; Schweighofer, Bernhard; Kupriyanova, Tatyana A et al. (2011) Tumor-recruited neutrophils and neutrophil TIMP-free MMP-9 regulate coordinately the levels of tumor angiogenesis and efficiency of malignant cell intravasation. Am J Pathol 179:1455-70
Fukuchi, Keisuke; Steiniger, Sebastian C J; Deryugina, Elena et al. (2010) Inhibition of tumor metastasis: functional immune modulation of the CUB domain containing protein 1. Mol Pharm 7:245-53
Deryugina, Elena I; Conn, Erin M; Wortmann, Andreas et al. (2009) Functional role of cell surface CUB domain-containing protein 1 in tumor cell dissemination. Mol Cancer Res 7:1197-211
Blouse, Grant E; Botkjaer, Kenneth A; Deryugina, Elena et al. (2009) A novel mode of intervention with serine protease activity: targeting zymogen activation. J Biol Chem 284:4647-57

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