Principal Investigator/Program Director (Last, first, middle): Kane, Lawrence, P ABSTRACT NF-?B family transcription factors are critical regulators of gene transcription. Mutations in the NF-?B pathway are now known to contribute to cellular transformation and the development of cancer. I have been studying the role of the serine/threonine kinase Akt in cellular activation and NF-?B induction. Interestingly, Akt is also a proto-oncogene, and is frequently amplified or activated in cancer. Akt is known to contribute to the activation of numerous downstream pathways, including NF-?B. I hypothesize that activation of NF-?B is important for the effects of Akt on T cell activation and transformation. At this point, the overall role of NF-?B in Akt-mediated transcriptional up-regulation is not clear. Also, it is not known precisely how Akt contributes to NF-?B activation. We have recently made some progress on this latter question, by showing that the protein CARMA1 is required for Akt-mediated NF-?B induction in T cells. Also, Akt can interact with CARMA1 and modulate its localization, in addition to increasing the phosphorylation of Bcl10, which is found in a complex with CARMA1. Based on our preliminary data and the hypothesis stated above, three specific aims are proposed. (1) To better understand how the interaction between Akt and CARMA1 affects cellular activation, we will employ a variety of molecular and cell biological approaches that will reveal in detail how this interaction is regulated. (2) To elucidate the role and regulation of Bcl10 phosphorylation, we will map sites of inducible phosphorylation within Bcl10 and determine both their functional relevance and the role of Akt in their regulation. (3) To determine the global role of NF-?B in Akt-mediated gene regulation and transformation, we will use microarray technology, first in conjunction with a powerful inhibitor of the NF-?B pathway, to reveal which Akt-inducible genes require NF-?B activity. Conversely, we will also inhibit Akt activity, and assess the effects on NF-?B regulated genes induced during T cell activation. Completion of these studies should yield important information about the cooperation between two important regulators of normal and neoplastic cell growth - Akt and NF-?B. Project Description Page 6

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
3R01GM080398-03S1
Application #
7882941
Study Section
Cellular Signaling and Dynamics Study Section (CSD)
Project Start
2009-09-01
Project End
2012-08-31
Budget Start
2009-09-01
Budget End
2012-08-31
Support Year
3
Fiscal Year
2009
Total Cost
$355,013
Indirect Cost
Name
University of Pittsburgh
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
004514360
City
Pittsburgh
State
PA
Country
United States
Zip Code
15213
Hawse, William F; Sheehan, Robert P; Miskov-Zivanov, Natasa et al. (2015) Cutting Edge: Differential Regulation of PTEN by TCR, Akt, and FoxO1 Controls CD4+ T Cell Fate Decisions. J Immunol 194:4615-9
Hamilton, Kristia S; Phong, Binh; Corey, Catherine et al. (2014) T cell receptor-dependent activation of mTOR signaling in T cells is mediated by Carma1 and MALT1, but not Bcl10. Sci Signal 7:ra55
Cheng, Jing; Hamilton, Kristia S; Kane, Lawrence P (2014) Phosphorylation of Carma1, but not Bcl10, by Akt regulates TCR/CD28-mediated NF-?B induction and cytokine production. Mol Immunol 59:110-6
Miskov-Zivanov, Natasa; Turner, Michael S; Kane, Lawrence P et al. (2013) The duration of T cell stimulation is a critical determinant of cell fate and plasticity. Sci Signal 6:ra97
DeFrances, Marie C; Debelius, Daniel R; Cheng, Jing et al. (2012) Inhibition of T-cell activation by PIK3IP1. Eur J Immunol 42:2754-9
Cheng, Jing; Montecalvo, Angela; Kane, Lawrence P (2011) Regulation of NF-?B induction by TCR/CD28. Immunol Res 50:113-7
Cheng, Jing; Phong, Binh; Wilson, David C et al. (2011) Akt fine-tunes NF-?B-dependent gene expression during T cell activation. J Biol Chem 286:36076-85
Ho, Allen W; Shen, Fang; Conti, Heather R et al. (2010) IL-17RC is required for immune signaling via an extended SEF/IL-17R signaling domain in the cytoplasmic tail. J Immunol 185:1063-70
Hu, Wei; Nessler, Stefan; Hemmer, Bernhard et al. (2010) Pharmacological prion protein silencing accelerates central nervous system autoimmune disease via T cell receptor signalling. Brain 133:375-88
Turner, Michael S; Kane, Lawrence P; Morel, Penelope A (2009) Dominant role of antigen dose in CD4+Foxp3+ regulatory T cell induction and expansion. J Immunol 183:4895-903