Mutations in genes associated with DNA mismatch repair (MMR) have been linked to hereditary colon cancer in humans. Transgenic mice in which the MMR genes have been inactivated are prone to cancer. However, the different knock-out mice develop different types of tumors, suggesting that the respective proteins may have non-overlapping roles in the preservation of genomic integrity and in tumor suppression. The principal investigator has developed a transgenic mouse assay system to study mutagenesis in vivo using a chromosomally-integrated, recoverable lambda phage shuttle vector carrying the bacterial supF gene as a mutation reporter. The broad, long-term objective of this proposal is to combine this in vivo mutation assay with the MMR-deficient animals to probe critical issues relating to genetic instability, carcinogenesis, and response to cancer therapy in the setting of MMR deficiency.
The specific aims are: 1) To generate second-generation transgenic mice with an improved supF reporter construct that is transcribed in mouse cells and that is modified to contain a signature sequence to identify sibling vs. independent mutations. 2) To examine spontaneous mutagenesis in hybrid mice, all carrying the XsupF shuttle vector, plus various combinations of alleles at the MMR loci, including PMS1, PMS2, MLH1, and MSH2. Each genotype will be compared for tissue-specific, age-related, and developmental differences. Mutation frequencies and spectra will be analyzed. 3) To examine induced mutagenesis in the mice by alkylating agents, UV light and base analogs in order to probe the role(s) of the MMR factors in cellular pathways other than mismatch correction, such as transcription-coupled repair. 4)To investigate the role of MMR in response to ionizing radiation, with emphasis on mutagenesis, cytotoxicity, and recognition of x-ray damage, based on the principal investigator`s preliminary data showing that MMR-deficient cells are x-ray resistant. 5) To test the role of MMR in suppressing genetic instability secondary to errors made by polymerase during base excision repair. Use will be made of transgenic mice with a targeted disruption of the polymerase locus, as well as the availability of a mutator variant of the polymerase to test the influence of MMR on polymerase -mediated mutagenesis.

Agency
National Institute of Health (NIH)
Institute
National Institute of Environmental Health Sciences (NIEHS)
Type
Research Project (R01)
Project #
5R01ES005775-08
Application #
6150673
Study Section
Chemical Pathology Study Section (CPA)
Program Officer
Velazquez, Jose M
Project Start
1992-09-30
Project End
2003-01-31
Budget Start
2000-02-01
Budget End
2001-01-31
Support Year
8
Fiscal Year
2000
Total Cost
$259,992
Indirect Cost
Name
Yale University
Department
Radiation-Diagnostic/Oncology
Type
Schools of Medicine
DUNS #
082359691
City
New Haven
State
CT
Country
United States
Zip Code
06520
Sulkowski, Parker L; Sundaram, Ranjini K; Oeck, Sebastian et al. (2018) Krebs-cycle-deficient hereditary cancer syndromes are defined by defects in homologous-recombination DNA repair. Nat Genet 50:1086-1092
Kim, Hoon; Lin, Qun; Glazer, Peter M et al. (2018) The hypoxic tumor microenvironment in vivo selects the cancer stem cell fate of breast cancer cells. Breast Cancer Res 20:16
Sulkowski, Parker L; Scanlon, Susan E; Oeck, Sebastian et al. (2018) PTEN Regulates Nonhomologous End Joining By Epigenetic Induction of NHEJ1/XLF. Mol Cancer Res 16:1241-1254
Lu, Yuhong; Liu, Yanfeng; Oeck, Sebastian et al. (2018) Hypoxia Promotes Resistance to EGFR Inhibition in NSCLC Cells via the Histone Demethylases, LSD1 and PLU-1. Mol Cancer Res 16:1458-1469
Scanlon, Susan E; Hegan, Denise C; Sulkowski, Parker L et al. (2018) Suppression of homology-dependent DNA double-strand break repair induces PARP inhibitor sensitivity in VHL-deficient human renal cell carcinoma. Oncotarget 9:4647-4660
Scanlon, Susan E; Scanlon, Christine D; Hegan, Denise C et al. (2017) Nickel induces transcriptional down-regulation of DNA repair pathways in tumorigenic and non-tumorigenic lung cells. Carcinogenesis 38:627-637
Gupta, Anisha; Quijano, Elias; Liu, Yanfeng et al. (2017) Anti-tumor Activity of miniPEG-?-Modified PNAs to Inhibit MicroRNA-210 for Cancer Therapy. Mol Ther Nucleic Acids 9:111-119
Sulkowski, Parker L; Corso, Christopher D; Robinson, Nathaniel D et al. (2017) 2-Hydroxyglutarate produced by neomorphic IDH mutations suppresses homologous recombination and induces PARP inhibitor sensitivity. Sci Transl Med 9:
Yu, Chang; Zelterman, Daniel (2017) A parametric model to estimate the proportion from true null using a distribution for p-values. Comput Stat Data Anal 114:105-118
Czochor, Jennifer R; Sulkowski, Parker; Glazer, Peter M (2016) miR-155 Overexpression Promotes Genomic Instability by Reducing High-fidelity Polymerase Delta Expression and Activating Error-Prone DSB Repair. Mol Cancer Res 14:363-73

Showing the most recent 10 out of 29 publications