PROJECT 3. A continuing programmatic goal is elucidation of mechanisms through which reactive nitrogen species (RNS) and reactive oxygen species (ROS) contribute to increased cancer risks. Project 3 addresses this goal by testing the hypothesis that damage to DMAand other cellular macromolecules by RNS from NO- producing macrophages and/or ROSfrom neutrophils either drives cells into apoptosis or inhibits apoptosis and enhances mutation. We employ models enabling mechanistic studies of DNA damage, genotoxicity, mutagenicity and cell death. Project 3 also defines DNA lesions with biological properties that explain mutagenic and lethal endpoints in cellular- and organism-level systems. Association of inflammatory bowel disease with risk of colon cancer is well-documented, and evidence also clearly associates high frequency of iNOS-containing tumor cells with poor survival of stage III melanoma patients. Thus, our first specific aim is to elucidate mechanisms underlying relationships among dose and dose-rate, DNA damage, mutagenesis and apoptosis induced in human colon carcinoma cells and human melanoma cells by exposure to RNS and ROS. The pivotal role of p53 in modulating responses will be evaluated by parallel studies in closely related p53-mutant and p53-null cells. Second, we shall characterize effects of dose and dose-rate on mutagenic potency and mutation spectra induced by RNS and ROS in the gpt reporter gene in three settings: (a) in pSV2gpt-transformed CHO AS52 cells exposed in vitro to NO* under controlled conditions;(b) in AS52 cells co-cultivated with activated RAW264.7 mouse macrophages and/or HL60 cells;and (c) integrated into the genome of Rag 2-1- IL10-/- mice developing inflammation-related colon adenocarcinoma.
Our third aim concerns genetic prioritization of DNA adducts as biomarkers of lethal and mutagenic endpoints. Adducts already nominated for potential as inflammation-derived pre-mutagenic lesions will first be evaluated by insertion into oligodeoxynucleotides, ligation into a viral genome and replication in E. coli cells of differing repair proficiency. Secondly we shall use the novel tool of chemical-biological fingerprinting to accelerate functional linkage of DNA damage and mutational spectra to lesions responsible for specific biological endpoints. Mutagenicity corresponding to features of mutational spectra in the gpt gene damaged with RNS or ROS (aim #2), will be characterized structurally (in collaboration with Project 2).

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Program Projects (P01)
Project #
5P01CA026731-31
Application #
8017419
Study Section
Special Emphasis Panel (ZCA1)
Project Start
Project End
Budget Start
2010-01-01
Budget End
2010-12-31
Support Year
31
Fiscal Year
2010
Total Cost
$367,263
Indirect Cost
Name
Massachusetts Institute of Technology
Department
Type
DUNS #
001425594
City
Cambridge
State
MA
Country
United States
Zip Code
02139
Gu, Chen; Ramos, Jillian; Begley, Ulrike et al. (2018) Phosphorylation of human TRM9L integrates multiple stress-signaling pathways for tumor growth suppression. Sci Adv 4:eaas9184
Wadduwage, Dushan N; Kay, Jennifer; Singh, Vijay Raj et al. (2018) Automated fluorescence intensity and gradient analysis enables detection of rare fluorescent mutant cells deep within the tissue of RaDR mice. Sci Rep 8:12108
Tajai, Preechaya; Fedeles, Bogdan I; Suriyo, Tawit et al. (2018) An engineered cell line lacking OGG1 and MUTYH glycosylases implicates the accumulation of genomic 8-oxoguanine as the basis for paraquat mutagenicity. Free Radic Biol Med 116:64-72
Rothenberg, Daniel A; Taliaferro, J Matthew; Huber, Sabrina M et al. (2018) A Proteomics Approach to Profiling the Temporal Translational Response to Stress and Growth. iScience 9:367-381
Wang, Xin; Garcia, Carlos T; Gong, Guanyu et al. (2018) Automated Online Solid-Phase Derivatization for Sensitive Quantification of Endogenous S-Nitrosoglutathione and Rapid Capture of Other Low-Molecular-Mass S-Nitrosothiols. Anal Chem 90:1967-1975
Chan, Cheryl; Pham, Phuong; Dedon, Peter C et al. (2018) Lifestyle modifications: coordinating the tRNA epitranscriptome with codon bias to adapt translation during stress responses. Genome Biol 19:228
Fedeles, Bogdan I (2017) G-quadruplex-forming promoter sequences enable transcriptional activation in response to oxidative stress. Proc Natl Acad Sci U S A 114:2788-2790
Townsend, Todd A; Parrish, Marcus C; Engelward, Bevin P et al. (2017) The development and validation of EpiComet-Chip, a modified high-throughput comet assay for the assessment of DNA methylation status. Environ Mol Mutagen 58:508-521
Kimoto, Takafumi; Kay, Jennifer E; Li, Na et al. (2017) Recombinant cells in the lung increase with age via de novo recombination events and clonal expansion. Environ Mol Mutagen 58:135-145
Edrissi, Bahar; Taghizadeh, Koli; Moeller, Benjamin C et al. (2017) N6-Formyllysine as a Biomarker of Formaldehyde Exposure: Formation and Loss of N6-Formyllysine in Nasal Epithelium in Long-Term, Low-Dose Inhalation Studies in Rats. Chem Res Toxicol 30:1572-1576

Showing the most recent 10 out of 361 publications