CEACAM1 is a homotypic cell adhesion molecule that plays a critical role in epithelial cell polarization, including lumen formation for breast and prostate epithelial cells when grown in 3D culture. Both long (72AA) and short (12 AA) cytoplasmic domain isoforms are produced by alternative mRNA splicing, with the short form predominant in most epithelial cells, and the long form predominant in T-cells. In addition, the CEACAM1 gene is silenced in most cancers, and in the case of colon cancer, it is silenced as early as pre-malignant hyperplastic polyps and adenomas. We have shown that peptides derived from the short form interact directly with actin, tropomyosin, calmodulin, and annexin 2, playing a role in interactions with the cytoskeleton, and that when phosphorylated on Thr and Ser residues, initiate a mitochondrial pathway of apoptosis, playing a role in lumen formation. In order to dissect these roles more fully, we propose to determine the hierarchal sequence of events that lead to productive interactions of the short form with the cytoskeleton, to identify the downstream kinases and adaptor proteins that lead to apoptosis by the short form, to determine the mechanism of receptor signaling inhibition by the long form, and to dissect the mechanism of gene silencing in prostate and colon cancers. To achieve the first three aims we propose biochemical and proteomic approaches that allow the direct identification of interacting proteins and the testing of these interactions in vivo in cells undergoing lumen formation using siRNA, confocal, and FRET approaches. In vivo foot printing and proteomic approaches will be used to identify the promoter complexes responsible for gene silencing in prostate and colon cell lines that do or do not express CEACAM1. Functional analyses of identified factors will be performed by factor depletion using siRNA approaches. These studies should provide a mechanistic insight into the function of CEACAM1 in a relevant biological process, namely lumen formation in normal differentiation, and the mechanism of CEACAM1 gene silencing in cancers of epithelial cell origin and the consequences thereof.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA084202-10
Application #
7822777
Study Section
Tumor Cell Biology Study Section (TCB)
Program Officer
Woodhouse, Elizabeth
Project Start
1999-12-01
Project End
2012-05-31
Budget Start
2010-06-09
Budget End
2012-05-31
Support Year
10
Fiscal Year
2010
Total Cost
$424,730
Indirect Cost
Name
City of Hope/Beckman Research Institute
Department
Type
DUNS #
027176833
City
Duarte
State
CA
Country
United States
Zip Code
91010
Weng, Chunyue; Nguyen, Tung; Shively, John E (2016) miRNA-342 Regulates CEACAM1-induced Lumen Formation in a Three-dimensional Model of Mammary Gland Morphogenesis. J Biol Chem 291:16777-86
Nguyen, Tung; Shively, John E (2016) Induction of Lumen Formation in a Three-dimensional Model of Mammary Morphogenesis by Transcriptional Regulator ID4: ROLE OF CaMK2D IN THE EPIGENETIC REGULATION OF ID4 GENE EXPRESSION. J Biol Chem 291:16766-76
Nguyen, Tung; Chen, Charng-Jui; Shively, John E (2014) Phosphorylation of CEACAM1 molecule by calmodulin kinase IID in a three-dimensional model of mammary gland lumen formation. J Biol Chem 289:2934-45
Zhang, Hui; Eisenried, Andreas; Zimmermann, Wolfgang et al. (2013) Role of CEACAM1 and CEACAM20 in an in vitro model of prostate morphogenesis. PLoS One 8:e53359
Li, Yun; Shively, John E (2013) CEACAM1 regulates Fas-mediated apoptosis in Jurkat T-cells via its interaction with ?-catenin. Exp Cell Res 319:1061-72
Samineni, Sridhar; Zhang, Zhifang; Shively, John E (2013) Carcinoembryonic antigen-related cell adhesion molecule 1 negatively regulates granulocyte colony-stimulating factor production by breast tumor-associated macrophages that mediate tumor angiogenesis. Int J Cancer 133:394-407
Lu, Rongze; Kujawski, Maciej; Pan, Hao et al. (2012) Tumor angiogenesis mediated by myeloid cells is negatively regulated by CEACAM1. Cancer Res 72:2239-50
Lu, Rongze; Pan, Hao; Shively, John E (2012) CEACAM1 negatively regulates IL-1ýý production in LPS activated neutrophils by recruiting SHP-1 to a SYK-TLR4-CEACAM1 complex. PLoS Pathog 8:e1002597
Chen, Lanfen; Chen, Zhangguo; Baker, Kristi et al. (2012) The short isoform of the CEACAM1 receptor in intestinal T cells regulates mucosal immunity and homeostasis via Tfh cell induction. Immunity 37:930-46
Li, Lin; Crow, Desiree; Turatti, Fabio et al. (2011) Site-specific conjugation of monodispersed DOTA-PEGn to a thiolated diabody reveals the effect of increasing peg size on kidney clearance and tumor uptake with improved 64-copper PET imaging. Bioconjug Chem 22:709-16

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