We previously demonstrated that AKT1 is the true human counterpart of v-akt and recently detected frequent elevated AKT1 kinase activity in primary tumors of prostate, breast and ovary. We have also observed that activated AKT1 transforms NIH 313 cells and induces membrane ruffling and that DN-AKT1 inhibits ras transformation. In addition, two AKI 1 -interation proteins have been identified, one of which is APciB that contains 4 AKT 1 phosphorylation sites and 3 SH3 domains, and the other is a proteasome b-subunit (PMSB4) that has an AKT1 phosphotylation site. AKT1 phosphorylates APaB and PSMB4 in vitro and in vivo. Moreover, APaB binds to PAK 1 kinase and enhances AKT1-induced cell survival and cell growth. We have also shown that AKI I phosphorylates p21 WAFI and abrogates p21 inhibitory effect on CDK4/6 and cdc2 kinases. Activated AKT1 overrides p5 3-induced Gi arrest. Based on these data, we hypothesize that that activated AKT1 and its associated proteins are cross-talk with PAK1 and p21 WAFi pathways to contribute to the control of malignant transformation, cell cycle progression and cytoskeleton reorganization. Thus, AKT1 could be a valuable tumor marker and a critical target for therapeutic intervention in human cancer. The broad, long-term objective of this project is to elucidate the normal cellular function of the AKT1 protein and determine the importance of perturbations of AKT1 pathway in human cancers.
The specific aims are: 1. Determine the clinicopathologic significance and the role of AKT1 activation in prostate and breast cancer. This will be accomplished by (a) examining AKT1 activation in a large series of primary prostate and breast carcinomas and correlating AKT1 activation with clinicopathologic features, and (b) creating prostate-specific, probasin promoter-driven AKTJ transgenic mice to ascertain if overexpression of activated AKT1 is tumorigenic. 2. Define the interaction between AKT1 and two potentially important signaling partners identified by the yeast two-hybrid system. We will examine (a) the effects of AKT 1-interaction proteins on AKT1 cell survival pathway, including PAK1 activation, Bad phosphorylation, NFiB activation, cytochrome c release, cytoskeleton reorganization, and p27 degradation, (b) the effects of AKT 1-interaction proteins on AKT1 oncogenic activity, and (c) the importance of the binding domains between AKT1 and its associated proteins in AKT1 signaling. 3. Examine the effects of AKT1 phosphorylation of p21 on cell cycle control. This will be accomplished by examining the effects of AKT1-induced p21 phosphorylation on (a) subcellular localization of p21 and p21 binding ability to cyclinlCDK complexes, (b) cyclinlCDK activity, (c) assembly of Gi cyclinlCDK complexes and PCNA binding activity, and (d) p21-induced cell cycle arrest. 4. Determine AKT1 as a critical target for cancer intervention. This will be accomplished by examining (a) the effects of direct inhibition of AKT1 on cancer cell growth, (b) the effects of DN-AKT1 and activated AKT1 on sensitivity of chemotherapeutic drugs, and (c) potential AKT1 inhibitors' specificity and ability to inhibit tumor cell growth in vitro and in vivo.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA089242-04
Application #
6748974
Study Section
Pathology B Study Section (PTHB)
Program Officer
Perry, Mary Ellen
Project Start
2001-06-08
Project End
2006-05-31
Budget Start
2004-06-01
Budget End
2005-05-31
Support Year
4
Fiscal Year
2004
Total Cost
$216,225
Indirect Cost
Name
University of South Florida
Department
Pathology
Type
Schools of Medicine
DUNS #
069687242
City
Tampa
State
FL
Country
United States
Zip Code
33612
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Fang, Quan; Naidu, K Akhilender; Naidu, Kamatham A et al. (2006) Ascorbyl stearate inhibits cell proliferation and tumor growth in human ovarian carcinoma cells by targeting the PI3K/AKT pathway. Anticancer Res 26:203-9
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Yuan, Zeng-qiang; Kim, Donghwa; Kaneko, Satoshi et al. (2005) ArgBP2gamma interacts with Akt and p21-activated kinase-1 and promotes cell survival. J Biol Chem 280:21483-90
Dan, Han C; Jiang, Kun; Coppola, Domenico et al. (2004) Phosphatidylinositol-3-OH kinase/AKT and survivin pathways as critical targets for geranylgeranyltransferase I inhibitor-induced apoptosis. Oncogene 23:706-15
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Fraser, Michael; Leung, Brendan M; Yan, Xiaojuan et al. (2003) p53 is a determinant of X-linked inhibitor of apoptosis protein/Akt-mediated chemoresistance in human ovarian cancer cells. Cancer Res 63:7081-8
Gritsko, Tatiana M; Coppola, Domenico; Paciga, June E et al. (2003) Activation and overexpression of centrosome kinase BTAK/Aurora-A in human ovarian cancer. Clin Cancer Res 9:1420-6
Yuan, Zeng-Qiang; Feldman, Richard I; Sussman, Gene E et al. (2003) AKT2 inhibition of cisplatin-induced JNK/p38 and Bax activation by phosphorylation of ASK1: implication of AKT2 in chemoresistance. J Biol Chem 278:23432-40
Sun, Mei; Yang, Lin; Feldman, Richard I et al. (2003) Activation of phosphatidylinositol 3-kinase/Akt pathway by androgen through interaction of p85alpha, androgen receptor, and Src. J Biol Chem 278:42992-3000

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