The long-term goal of the proposed research project is to understand the molecular mechanisms of cellular responses to a variety of stresses in mammalian cells. During the next granting period, we will investigate the effect of metabolic oxidative stress on TRAIL (tumor necrosis factor-related apoptosis-inducing ligand)-induced apoptotic death. We postulate that tumor microenvironment, in particular glucose deprivation, enhances TRAIL-induced cytotoxicity by facilitating the mitochondria-dependent pathway and/or -independent pathway. Low glucose concentrations induce metabolic oxidative stress and subsequently promote cytochrome c release in the presence of TRAIL. Cytochrome c release promotes the mitochondria-mediated caspase signal transduction pathway. We also hypothesize that metabolic oxidative stress-induces an elevation of ceramide which activates ceramide-activated protein phosphatase (CAPP) and/or inactivates phosphoinositide-3 kinase [PI(3)K]. The activation of CAPP and/or inactivation of PI(3)K inactivates Akt through dephosphorylation and consequently down-regulates the expression of FLIP, an antiapoptotic protein, by inhibiting NF-kappaB signal transduction pathway.
The specific aims of this project are to examine (1) the effect of low glucose concentrations on TRAIL-induced cytotoxicity, (2) the effect of low glucose concentrations on the TRAIL-activated apoptotic pathway, (3) cross-talk between the mitochondria-dependent caspase pathway and the apoptosis associated protein-regulated caspase pathway, (4) the role of the metabolic oxidative stress-ceramide-PI(3)K/CAPP-Akt-NF-kappaB-FLIP pathway in TRAIL sensitivity, (5) the effect of a variety of other characteristic features of the tumor microenvironment on TRAIL cytotoxicity. The proposed studies (Aim 1) will use survival determination assays to examine TRAIL-induced cytotoxicity in various concentrations of glucose.
The second aim will focus on the use of biochemical approaches to investigate how low glucose concentrations promote the TRAIL-activated caspase signal transduction pathway. The studies for Aim 3 will use molecular and biochemical approaches to elucidate cross-talk between the mitochondria-dependent caspase pathway and apoptosis associated protein-regulated caspase pathway. We will employ molecular genetics and biochemical techniques to elucidate the role of the PI(3)K/CAPP-Akt-FLIP pathway in the low glucose-enhanced TRAIL cytotoxicity (Aim 4). Finally we will integrate different aspects of the signaling pathways. We believe that the successful outcome of this study will support the development and clinical application of TRAIL for the treatment of human cancer.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
1R01CA095191-01A2
Application #
6679108
Study Section
Metabolic Pathology Study Section (MEP)
Program Officer
Spalholz, Barbara A
Project Start
2003-07-01
Project End
2008-06-30
Budget Start
2003-07-01
Budget End
2004-06-30
Support Year
1
Fiscal Year
2003
Total Cost
$265,220
Indirect Cost
Name
University of Pittsburgh
Department
Surgery
Type
Schools of Medicine
DUNS #
004514360
City
Pittsburgh
State
PA
Country
United States
Zip Code
15213
Song, Jae J; Szczepanski, Miroslaw Jerzy; Kim, So Young et al. (2010) c-Cbl-mediated degradation of TRAIL receptors is responsible for the development of the early phase of TRAIL resistance. Cell Signal 22:553-63
Alcala Jr, Marco A; Park, Kyungsoo; Yoo, Jinsang et al. (2010) Effect of hyperthermia in combination with TRAIL on the JNK-Bim signal transduction pathway and growth of xenograft tumors. J Cell Biochem 110:1073-81
Lee, Dae-Hee; Rhee, Juong G; Lee, Yong J (2009) Reactive oxygen species up-regulate p53 and Puma; a possible mechanism for apoptosis during combined treatment with TRAIL and wogonin. Br J Pharmacol 157:1189-202
Lee, Dae-Hee; Szczepanski, Miroslaw-Jerzy; Lee, Yong J (2009) Magnolol induces apoptosis via inhibiting the EGFR/PI3K/Akt signaling pathway in human prostate cancer cells. J Cell Biochem 106:1113-22
Jeong, Jae-Hoon; An, Jee Young; Kwon, Yong Tae et al. (2009) Effects of low dose quercetin: cancer cell-specific inhibition of cell cycle progression. J Cell Biochem 106:73-82
Kim, Young-Ho; Lee, Dae-Hee; Jeong, Jae-Hoon et al. (2008) Quercetin augments TRAIL-induced apoptotic death: involvement of the ERK signal transduction pathway. Biochem Pharmacol 75:1946-58
Lee, Dae-Hee; Kim, Clifford; Zhang, Lin et al. (2008) Role of p53, PUMA, and Bax in wogonin-induced apoptosis in human cancer cells. Biochem Pharmacol 75:2020-33
Lee, Dae-Hee; Lee, Yong J (2008) Quercetin suppresses hypoxia-induced accumulation of hypoxia-inducible factor-1alpha (HIF-1alpha) through inhibiting protein synthesis. J Cell Biochem 105:546-53
Park, Sung-Soo; Bae, Insoo; Lee, Yong J (2008) Flavonoids-induced accumulation of hypoxia-inducible factor (HIF)-1alpha/2alpha is mediated through chelation of iron. J Cell Biochem 103:1989-98
Lee, Dae-Hee; Szczepanski, Miroslaw; Lee, Yong J (2008) Role of Bax in quercetin-induced apoptosis in human prostate cancer cells. Biochem Pharmacol 75:2345-55

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