Project Summary (2022-2027)

N-Nitrosamines include some of the most mutagenic chemicals known to humankind, and they are present at and near Superfund Sites, in drinking water, in food and in drugs. Given their ubiquity and potent mutagenicity, research focused on N-nitrosamines is critical. What is needed are better ways to predict N-nitrosamine-induced disease so that intervention strategies can be developed. A key barrier has been the difficulty of studying long-term, low-dose conditions. To overcome this challenge, this project creates “canary in a coal mine” genetically engineered model (C-GEM) mice. The C-GEM mice harbor genetic mutations in the alkyladenine DNA glycosylase (Aag) and the O6-methylguanine methyltransferase (Mgmt), two of the most important repair activities for NMDA-induced DNA damage. They also harbor two transgenes for detecting mutations (analyzed in collaboration with High Resolution Mutation Spectra and Multi-Omics for Deducing Etiology and Predicting Disease project). C-GEM mice are exposed to environmentally relevant levels of NDMA, determined by the Methods for Selective Extraction, Concentration, and Detection of N-Nitrosamines and Measurement and Engineering Solutions to Detect and Prevent N-Nitrosamine Exposure projects, and systems-level data (to be collected with the High Resolution Mutation Spectra and Multi-Omics for Deducing Etiology and Predicting Disease project) will be merged by the Data Management and Analysis Core (DMAC) to create predictive biomarkers and to better understand mechanisms of disease.

This project is also studying a probiotic organism shown to suppress NDMA-induced cancer. Finally, project researchers are creating “Chem-Sense Cells,” a novel cell array-based platform for screening environmentally relevant N-nitrosamines (to be identified by Methods for Selective Extraction, Concentration, and Detection of N-Nitrosamines, as well as Measurement and Engineering Solutions to Detect and Prevent N-Nitrosamine Exposure) for their DNA damaging potential. Aim 1 is to leverage the C-GEM mice at a sensitive window of susceptibility to reveal the long-term, low-dose impact of NDMA administered under environmentally relevant conditions. Aim 2 is to use chemical-genetics to create Chem-Sense Cells as a novel biosensor for DNA damage induced by N-nitrosamines present at Superfund Sites. Aim 3 is to leverage the C-GEM mice to reveal the biological mechanisms by which probiotics modulate mutation susceptibility via studies of systems-level responses.

Trainees drive the research and are supported by the Research Experience and Training Coordination Core to ensure optimal professional development. With support from the Community Engagement Core, project leaders and trainees listen to and work with community members and tribes to develop novel hands-on teaching curricula that explain key concepts in biology that underlie their research activities. Importantly, this project is part of a greater Systems Approach for the entire Massachusetts Institute of Technology Superfund Research Program, for which interactions and interdependencies (supported by the Administrative Core) give rise to impact that is not otherwise possible. This includes using machine learning (in collaboration with the DMAC) to merge research results from this project with that of other projects to gain a better understanding of risk. The Administrative Core also promotes the dissemination of this project’s innovative mouse and cell technologies, and it supports bringing research results to the EPA and other stakeholders to maximize impact on public health.