Human tumor cells invade the extracellular matrix using plasma membrane protrusions, invadopodia, that contact and dissolve the matrix. A transiently expressed, type II transmembrane serine protease, seprase, and its protein complexes function on invadopodia. The goal of this proposal is to investigate the regulation of these invadopodia-associated proteins in development of the invasive and metastatic phenotypes at the levels of protein interaction and gene expression. Malignant human melanoma and ovarian carcinoma cells have definable invadopodia profiles; they are primed for survival in the tumor microenvironment and for subsequent spontaneous metastasis.
First Specific Aim focuses on the analysis of the role of these invadopodia-associated proteases and integrins in cell survival in the tumor microenvironment, and in tumor cell invasion and metastasis. The specific importance of invadopodia proteins will be defined using RNA interference (RNAi) knockdown of mRNAs in invasive tumor cells to destroy the invadopodia-associated adhesive and proteolytic activities.
Second Aim assesses the mechanism of seprase activation in the context of expression of active domains of the enzyme and identification of peptide and antibody inhibitors.
Third Aim will identify molecules important in progression of ovarian cancer from primary tumor to ascites. Tumor cells will be isolated from primary tumor and ascites of patients with ovarian cancer; their gene expression profiles will be determined using DNA microarray and real-time RT-PCR; their cell invasiveness in vitro and tumor growth and metastasis potential in vivo will be demonstrated. Specific genes upregulated in invasive tumor cells will be validated using the RNAi knockdown approach.
Fourth Aim i s to discover natural substrates for seprase and MT1-MMP by proteomic analysis on immuno-affinity-purified invadopodia complexes. Finally, using a novel animal model system, seprase-induced immune components implicated in the anti-tumor and anti-metastasis control will be identified. Thus, this proposal will analyze molecular basis of the invasive and metastatic phenotypes, with the eventual goal of determining invadopodia proteins as therapeutic and diagnostic targets for metastatic disease. ? ?

National Institute of Health (NIH)
National Cancer Institute (NCI)
Research Project (R01)
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Special Emphasis Panel (ZRG1-CPA (03))
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Ault, Grace S
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State University New York Stony Brook
Internal Medicine/Medicine
Schools of Medicine
Stony Brook
United States
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Tulley, Shaun; Chen, Wen-Tien (2014) Transcriptional regulation of seprase in invasive melanoma cells by transforming growth factor-? signaling. J Biol Chem 289:15280-96
Javidroozi, Mazyar; Zucker, Stanley; Chen, Wen-Tien (2012) Plasma seprase and DPP4 levels as markers of disease and prognosis in cancer. Dis Markers 32:309-20
Lu, Janice; Fan, Tina; Zhao, Qiang et al. (2010) Isolation of circulating epithelial and tumor progenitor cells with an invasive phenotype from breast cancer patients. Int J Cancer 126:669-83
Fan, Tina; Zhao, Qiang; Chen, John J et al. (2009) Clinical significance of circulating tumor cells detected by an invasion assay in peripheral blood of patients with ovarian cancer. Gynecol Oncol 112:185-91
Kennedy, Alanna; Dong, Huan; Chen, Donghai et al. (2009) Elevation of seprase expression and promotion of an invasive phenotype by collagenous matrices in ovarian tumor cells. Int J Cancer 124:27-35
Freudenberg, Jaclyn A; Chen, Wen-Tien (2007) Induction of Smad1 by MT1-MMP contributes to tumor growth. Int J Cancer 121:966-77
Ghersi, Giulio; Zhao, Qiang; Salamone, Monica et al. (2006) The protease complex consisting of dipeptidyl peptidase IV and seprase plays a role in the migration and invasion of human endothelial cells in collagenous matrices. Cancer Res 66:4652-61
Chen, Donghai; Kennedy, Alanna; Wang, Jaw-Yuan et al. (2006) Activation of EDTA-resistant gelatinases in malignant human tumors. Cancer Res 66:9977-85
Chen, Wen-Tien; Kelly, Thomas (2003) Seprase complexes in cellular invasiveness. Cancer Metastasis Rev 22:259-69
Chen, Wen-Tien; Kelly, Thomas; Ghersi, Giulio (2003) DPPIV, seprase, and related serine peptidases in multiple cellular functions. Curr Top Dev Biol 54:207-32

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