We have developed orthotopic xenograft transplants from human colon cancer cell lines that recapitulate the metastatic pattern of colon cancer in patients and have made the novel observation that rescue of attenuated TGF? receptor expression inhibits metastatic capability in these models even though invasion at the primary tumor site was maintained. Moreover, we showed genetic attenuation of TGF? receptor expression in a model that was poorly metastatic generated robust metastasis to the liver and lungs. These results are significant because they raise the possibility that regeneration of attenuated TGF? signaling would be an effective target in the treatment of metastases. Development of effective treatments for metastatic disease would have a large impact since metastases are the major cause of death in solid cancers. A key to capitalizing on the repression of metastases TGF? signaling is the elucidation of the mechanism by which metastasis is inhibited. The mechanistic basis of this project is our novel observation that TGF? mediates control of cell death through a mechanism involving the activation of PKA (independent of cAMP, but AKAP dependent) that leads to proteosomal degradation of survivin and XIAP. These IAPs have been shown to be critical to aberrant cell survival signaling in tumor models (Dohi et al, 2007). PKA initiates a multi-functional cascade directed at disruption of this cell survival mechanism.
Aims I and II will focus on the characterization of this pathway.
Specific Aim I will address the hypothesis that Smad 2 plays a critical role in recruitment of the survivin/XIAP complex to the vicinity of the activated PKA. There are two PKA regulatory subunit isoforms designated PKAI and PKAII. We have found that poorly metastatic cells have a preponderance of RII while the isogenic complementary highly metastatic cells have high RI. This suggests that the regulatory subunits function differently from each other with respect to the efficiency of TGF?/PKA pathway activation and that TGF? signaling affects the expression of the isoforms as well. Thus these 2 hypotheses will also be tested in Aim I. Our previous work noted that in addition to repression of survivin, AKT was inhibited by TGF? signaling. Since AKT dephosphorylation is carried out by PP2A (another AKAP directed enzyme) and is a PKA substrate, the hypothesis that PP2A activation is part of the TGF?/PKA pathway and plays a dual role countering cell survival through promotion of proteasomal stimulation and ensuing degradation of XIAP will be tested in Aim II. vivo. Due to the critical role of the survivin/XIAP complex in controlling cell survival in vitro, we will test the hypothesis that mutations which stabilize the complex will enhance metastatic colonization in poorly metastatic cells and to block regression of established metastases by TGF? receptor reconstitution in highly metastatic cells in Specific Aim III.
The specific aims of the project are: I. Determine the mechanism of TGF? activation of PKA;II. Determine the potential role of PP2A in the TGF?/PKA pathway. III. Test the hypothesis that the survivin/XIAP complex is essential to metastatic spread.
One of the most pressing problems in cancer treatment is the lack of effective therapies for advanced metastatic disease. The elucidation of metastatic mechanisms and the validation that their disruption results in the regression of the metastases will be addressed in this project. We have identified a candidate mechanism for inappropriate cell survival signaling in metastases that will be subjected to validation and evaluation as a therapeutic target for the treatment of metastases in this project.
|Leiphrakpam, Premila D; Rajput, Ashwani; Mathiesen, Michelle et al. (2014) Ezrin expression and cell survival regulation in colorectal cancer. Cell Signal 26:868-79|
|Leiphrakpam, Premila D; Agarwal, Ekta; Mathiesen, Michelle et al. (2014) In vivo analysis of insulin-like growth factor type 1 receptor humanized monoclonal antibody MK-0646 and small molecule kinase inhibitor OSI-906 in colorectal cancer. Oncol Rep 31:87-94|
|Agarwal, Ekta; Chaudhuri, Anathbandhu; Leiphrakpam, Premila D et al. (2014) Akt inhibitor MK-2206 promotes anti-tumor activity and cell death by modulation of AIF and Ezrin in colorectal cancer. BMC Cancer 14:145|
|Chowdhury, Sanjib; Ongchin, Melanie; Sharratt, Elizabeth et al. (2013) Intra-tumoral heterogeneity in metastatic potential and survival signaling between iso-clonal HCT116 and HCT116b human colon carcinoma cell lines. PLoS One 8:e60299|
|Agarwal, Ekta; Brattain, Michael G; Chowdhury, Sanjib (2013) Cell survival and metastasis regulation by Akt signaling in colorectal cancer. Cell Signal 25:1711-9|
|Chowdhury, Sanjib; Ongchin, Melanie; Wan, Guanghua et al. (2013) Restoration of PTEN activity decreases metastases in an orthotopic model of colon cancer. J Surg Res 184:755-60|
|Hedrick, Erik D; Agarwal, Ekta; Leiphrakpam, Premila D et al. (2013) Differential PKA activation and AKAP association determines cell fate in cancer cells. J Mol Signal 8:10|
|Zou, Yi; Howell, Gillian M; Humphrey, Lisa E et al. (2013) Ron knockdown and Ron monoclonal antibody IMC-RON8 sensitize pancreatic cancer to histone deacetylase inhibitors (HDACi). PLoS One 8:e69992|
|Sabbah, Dima A; Simms, Neka A; Wang, Wang et al. (2012) N-Phenyl-4-hydroxy-2-quinolone-3-carboxamides as selective inhibitors of mutant H1047R phosphoinositide-3-kinase (PI3KÃ½Ã½). Bioorg Med Chem 20:7175-83|
|Sabbah, Dima A; Simms, Neka A; Brattain, Michael G et al. (2012) Biological evaluation and docking studies of recently identified inhibitors of phosphoinositide-3-kinases. Bioorg Med Chem Lett 22:876-80|
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