The long-range goals of this grant proposal are to study the involvement of the hypoxia signaling pathway upon chromatin remodeling, gene silencing, and cell transformation. Hypoxia or agents that mimic hypoxia cause a global loss of histone acetylation and increase the methylation of H3 lysine 9 which is the chromatin mark of gene silencing for the individual nucleosomes. Methylation of H3 lysine 9 leads to DNA methylation and inherited gene silencing. We will study the ability of hypoxia and agents that activate hypoxia signaling (NiCI2, CoCI2, deferoxamine, and DMOG) to induce cell transformation in wild-type or in HIF-1alpha knockout mouse embryo fibroblast (MEF). We will transfect a normal HIF-1alpha construct into MEF cells with a knockout of this gene to study whether we can restore their ability to be transformed by hypoxia and agents that mimic hypoxia. We will also stably transfect mutated HIF-1alpha constructs that will constitutively express HIF in its stabile and or stable/active form into MEF HIF+/- cells and study the effect of stabilized and/or activated HIF- 1alpha on anchorage-independent growth and tumor formation in nude mice. We will investigate global changes in H3K9 acetylation, H3K9 methylation, G9a activity (enzyme that dimethylates H3K9) and DNA methylation in MEF with intact or knockout of HIF-1alpha following exposure to hypoxia or agents that mimic hypoxia. We will also utilize the expression levels of DHFR and DNA mismatch repair gene Mlh1 which are down-regulated by hypoxia signaling to study gene specific effects of chromatin remodeling induced by hypoxia signaling using CHIP assays targeted to their promoters with antibodies specific to H3K9 dimethylation and/or H3K9 acetylation. We have obtained mouse embryonic stem cell wild-type (WT), G9a knockout (G9a-/-) and G9a-/- stably transfected with wild-type G9a (G9a-/- + G9a WT), and we will utilize these cells to address the role of G9a in mediating histone methylation H3K9 and the effects of hypoxia signaling on DNA methylation and DHFR/MIh expression. We will study the reversibility and stability of MEF cells that have already been transformed via the hypoxia pathway to chromatin remodeling agents and R59949 (HIF inhibitor). These studies will allow us to understand the importance of hypoxia signaling and the HIF-1alpha transcription factor in promoting neoplastic cell transformation. ? ? ?

Agency
National Institute of Health (NIH)
Institute
National Institute of Environmental Health Sciences (NIEHS)
Type
Research Project (R01)
Project #
1R01ES014454-01A1
Application #
7138704
Study Section
Cancer Etiology Study Section (CE)
Program Officer
Tyson, Frederick L
Project Start
2006-08-16
Project End
2010-06-30
Budget Start
2006-08-16
Budget End
2007-06-30
Support Year
1
Fiscal Year
2006
Total Cost
$401,375
Indirect Cost
Name
New York University
Department
Public Health & Prev Medicine
Type
Schools of Medicine
DUNS #
121911077
City
New York
State
NY
Country
United States
Zip Code
10016
Brocato, Jason; Hernandez, Michelle; Laulicht, Freda et al. (2015) In Vivo Exposures to Particulate Matter Collected from Saudi Arabia or Nickel Chloride Display Similar Dysregulation of Metabolic Syndrome Genes. J Toxicol Environ Health A 78:1421-36
Brocato, Jason; Chen, Danqi; Liu, Jianli et al. (2015) A Potential New Mechanism of Arsenic Carcinogenesis: Depletion of Stem-Loop Binding Protein and Increase in Polyadenylated Canonical Histone H3.1 mRNA. Biol Trace Elem Res 166:72-81
Brocato, Jason; Costa, Max (2015) SATB1 and 2 in colorectal cancer. Carcinogenesis 36:186-91
Brocato, Jason; Wu, Fen; Chen, Yu et al. (2015) Association between sleeping hours and cardiometabolic risk factors for metabolic syndrome in a Saudi Arabian population. BMJ Open 5:e008590
Niu, Yingmei; DesMarais, Thomas L; Tong, Zhaohui et al. (2015) Oxidative stress alters global histone modification and DNA methylation. Free Radic Biol Med 82:22-8
Brocato, Jason; Chervona, Yana; Costa, Max (2014) Molecular responses to hypoxia-inducible factor 1? and beyond. Mol Pharmacol 85:651-7
Brocato, Jason; Fang, Lei; Chervona, Yana et al. (2014) Arsenic induces polyadenylation of canonical histone mRNA by down-regulating stem-loop-binding protein gene expression. J Biol Chem 289:31751-64
Brocato, Jason; Costa, Max (2013) Basic mechanics of DNA methylation and the unique landscape of the DNA methylome in metal-induced carcinogenesis. Crit Rev Toxicol 43:493-514
Arita, Adriana; Muñoz, Alexandra; Chervona, Yana et al. (2013) Gene expression profiles in peripheral blood mononuclear cells of Chinese nickel refinery workers with high exposures to nickel and control subjects. Cancer Epidemiol Biomarkers Prev 22:261-9
Passantino, Lisa; Muñoz, Alexandra B; Costa, Max (2013) Sodium metavanadate exhibits carcinogenic tendencies in vitro in immortalized human bronchial epithelial cells. Metallomics 5:1357-67

Showing the most recent 10 out of 39 publications