A major problem in cancer chemotherapy is intrinsic or acquired drug resistance. We recently found, for the first time, that the loss or decreased expression of beta2- miocroglobulin (beta2M) is involved in the development of drug resistance. This is a completely novel drug resistance mechanism which has not been previously described in the literature. Therefore, the overall goals of this project are (1) to unravel the molecular and biochemical mechanisms by which beta2M causes drug resistance, and (2) to develop strategies to circumvent drug resistance due to the loss of decreased expression of beta2m.
The Specific Aims are to (1) determine whether cellular levels of beta2m influence the cell cycle; (2) determine whether beta2m plays a role in apoptosis in drug sensitive cells, and whether its loss or decreased expression prevents apoptotic cell death; (3) modulate the expression of beta2m by cytokines and thereby circumvent drug resistance; and (4) investigate the regulation of beta2m expression in drug resistant cells, and determine the molecular mechanisms of beta2m gene suppression in these cells. In order to accomplish Specific Aim 1, experiments will be conducted to and decreases or increases cell proliferation, respectively, and (2) whether beta2m modulates the function of specific cell of specific cell-cycle, and decreases or increases cell proliferation, respectively, and (b) whether beta2m modulates the function of specific cell cycle-controlling proteins. To pursue Specific Aim 2, the role of beta2m in apoptosis will be explored by (a) evaluating levels in cells transfected with the beta2m gene in the sense or antisense orientation, and (b) examine whether anti-beta2m monoclonal antibodies in the absence or presence of chemotherapeutic agents, induce apoptosis in drug sensitive, but not in beta2m-deficient, cells. In order to accomplish Specific Aim 3, experiments will be conduced to evaluate the modulating effects of interferon-gamma (IFN- gamma) and tumor necrosis factor alpha (TNF-alpha), with or without doxorubican or vincristine, on beta2m expression in cells with reduced beta2m expression and in beta2m transfectants.
In Specific Aim 4, we will determine whether (a) IFN-gamma or TNF-alpha induces specific transcription factors to enhance transcription of the beta2m gene in drug resistant cells, and (b) determine whether reduced expression of beta2m in resistant cells is due to decreased or absent positive regulatory transcription factors, or the presence of transcription suppressors. These studies will aid in understanding the molecular mechanism(s) of this novel drug resistance phenotype due to the loss or decreased expression of beta2m, and will be useful for the development of more effective chemotherapeutic or potential gene therapy strategies.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
3R01CA080734-04S1
Application #
6642972
Study Section
Experimental Therapeutics Subcommittee 1 (ET)
Program Officer
Yovandich, Jason L
Project Start
1999-03-03
Project End
2003-02-28
Budget Start
2001-03-01
Budget End
2003-02-28
Support Year
4
Fiscal Year
2002
Total Cost
$74,931
Indirect Cost
Name
Indiana University-Purdue University at Indianapolis
Department
Pharmacology
Type
Schools of Medicine
DUNS #
005436803
City
Indianapolis
State
IN
Country
United States
Zip Code
46202
Safa, Ahmad R (2016) Resistance to Cell Death and Its Modulation in Cancer Stem Cells. Crit Rev Oncog 21:203-219
Safa, Ahmad R (2013) Roles of c-FLIP in Apoptosis, Necroptosis, and Autophagy. J Carcinog Mutagen Suppl 6:
Safa, A R (2012) c-FLIP, a master anti-apoptotic regulator. Exp Oncol 34:176-84
Safa, Ahmad R; Pollok, Karen E (2011) Targeting the Anti-Apoptotic Protein c-FLIP for Cancer Therapy. Cancers (Basel) 3:1639-71
Park, Soo-Jung; Bijangi-Vishehsaraei, Khadijeh; Safa, Ahmad R (2010) Selective TRAIL-triggered apoptosis due to overexpression of TRAIL death receptor 5 (DR5) in P-glycoprotein-bearing multidrug resistant CEM/VBL1000 human leukemia cells. Int J Biochem Mol Biol 1:90-100
Choi, Mi-Ran; Najafi, Farhad; Safa, Ahmad R et al. (2008) Analysis of changes in the proteome of HL-60 promyeloid leukemia cells induced by the proteasome inhibitor PSI. Biochem Pharmacol 75:2276-88
Safa, Ahmad R; Day, Travis W; Wu, Ching-Huang (2008) Cellular FLICE-like inhibitory protein (C-FLIP): a novel target for cancer therapy. Curr Cancer Drug Targets 8:37-46
Zhong, Xiaoling; Safa, Ahmad R (2007) Phosphorylation of RNA helicase A by DNA-dependent protein kinase is indispensable for expression of the MDR1 gene product P-glycoprotein in multidrug-resistant human leukemia cells. Biochemistry 46:5766-75
Park, Soo-Jung; Wu, Ching-Huang; Choi, Mi-Ran et al. (2006) P-glycoprotein enhances TRAIL-triggered apoptosis in multidrug resistant cancer cells by interacting with the death receptor DR5. Biochem Pharmacol 72:293-307
Day, Travis W; Najafi, Farhad; Wu, Ching-Huang et al. (2006) Cellular FLICE-like inhibitory protein (c-FLIP): a novel target for Taxol-induced apoptosis. Biochem Pharmacol 71:1551-61

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