A role of the toleromerase catalytic subunit, hTERT, in preventing telomeric attrition associated with cellular senescence has been established and recent studies have shown that hTERT is controlled primarily through transcriptional mechanisms. Considerable excitement has been generated by recent findings that hTERT expression may be induced in telomerase-negative cells through changes in histone acetylation and/or DNA methylation of the hTERT promoter. Since hTERT gene expression is inactivated during embryogenesis and remains transcriptionally silent in most somatic aging cells, these findings are of intense interest not only to those concerned with the causes of aging, but also to oncologists as hTERT is activated in most human cancers. Our hypothesis is that histone deacetylation and DNA methylation work in conjunction in embryonic cellular differentiation in embryonic cellular differentiation to silence the hTERT promoter and that this mechanism is attenuated in aging cells leading to a higher probability of hTERT activation and neoplasia with aging. To test this hypothesis, we have designed studies to 1) subject differentiating embryonic cells to trichostatin A and/or 5-azacytidine, which induce histone acetylation and hypomethylation of the hTERT promoter respectively, and assess hTERT activity in these cells, 2) treat young and senescent WI-38 fibroblasts with these agents to analyze the ability of histone deacetylation and/or DNA methylation to maintain hTERT gene silencing in aging cells, and 3) assess hTERT promoter regulation in differentiating embryonic cells and aging fibroblasts transfected with luciferase reporter constructs and subjected to histone acetylation and/or DNA hypomethylation. The overall purpose of this study is to further elucidate the mechanisms of histone acetylation and DNA methylation in the control of telomerase in aging and cancer. These studies are intended to strengthen the potential for development of therapeutic control of hTERT expression during aging and in age-associated diseases such as cancer.

Agency
National Institute of Health (NIH)
Institute
National Institute on Aging (NIA)
Type
Small Research Grants (R03)
Project #
1R03AG020375-01
Application #
6439871
Study Section
National Institute on Aging Initial Review Group (NIA)
Program Officer
Sierra, Felipe
Project Start
2002-09-01
Project End
2003-08-31
Budget Start
2002-09-01
Budget End
2003-08-31
Support Year
1
Fiscal Year
2002
Total Cost
$72,479
Indirect Cost
Name
University of Alabama Birmingham
Department
Biology
Type
Schools of Arts and Sciences
DUNS #
004514360
City
Birmingham
State
AL
Country
United States
Zip Code
35294
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Casillas, Mark A; Brotherton, Scott L; Andrews, Lucy G et al. (2003) Induction of endogenous telomerase (hTERT) by c-Myc in WI-38 fibroblasts transformed with specific genetic elements. Gene 316:57-65
Saldanha, Sabita N; Andrews, Lucy G; Tollefsbol, Trygve O (2003) Analysis of telomerase activity and detection of its catalytic subunit, hTERT. Anal Biochem 315:1-21
Lopatina, Nadejda G; Poole, Joseph C; Saldanha, Sabita N et al. (2003) Control mechanisms in the regulation of telomerase reverse transcriptase expression in differentiating human teratocarcinoma cells. Biochem Biophys Res Commun 306:650-9
Saldanha, Sabita N; Andrews, Lucy G; Tollefsbol, Trygve O (2003) Assessment of telomere length and factors that contribute to its stability. Eur J Biochem 270:389-403