A Diversity Outbred ES Cell Panel for In Vitro Genetics of Toxicology

DESCRIPTION provided by applicant Strain background can strongly influence the outcome of toxicity tests in animals Time and cost constraints preclude in vivo approaches to longstanding concerns about the lack of genetic diversity in animal models An in vitro platform is vastly more efficient for understanding the role of genetic background in toxicology testing and sacrifices no animals In this project we propose to build a large panel of genetically diverse ES cells from the Diversity Outcross line of mice The DO mice are an advanced intercross of the founder strains used for the Collaborative Cross The DO strains capture over of all sequence variants present in laboratory strains and harbor They are ideally suited for complex trait mapping ES cells offer tremendous flexibililty and is also ideal for an in vitro genetics platform DO ES cell lines immortalize DO genomes creating a renewable resource In principle ES cells can provide access to almost any cell type by directed differentiation In this way the DO ES panel is extremely versatile as a permanent renewable resource In Phase I we demonstrated feasibility by deriving a small set of ES cell lines and identifying a cytotoxicity QTL by in vitro genetic analysis Since then we have derived an additional DO ES cell lines and in Phase II we will validate and define screening panels of male and female DO ES cell lines that are well powered for QTL mapping We will biologically test candidate genes to identify at least mouse QTL genes and then determine the fraction of those genes that are also modifers of cytotoxicity in human IPS cells The key challenge to broad adoption of our technology is a demostration of the relevance of mouse QTL genes to human toxicological response The aims of this project are not primarily directed toward building the DO ES cell lines They are directed to validating their utility toward understanding mechanisms underlying variable susceptibility to toxicants in human cells While there are many cell types and toxicities that can be approached using our technology we focused this project on cytotoxicity in pluripotent stem cells to efficiently generate a well powered dataset to validat our technology The Phase II project will establish our in vitro genetics platform as an important and novel advance in understanding human toxicological response PUBLIC HEALTH RELEVANCE There is a longstanding interest in how genetics impacts toxicology but cost effective highly scalable tools to investigate this relationship are not available This project will improve human health and reduce the risk of environmental toxicants by developing a high throughput in vitro platform for investigating the genetic basis of variable response to toxicant exposure