CRISPR screens of population relevant genes governing toxicant resilience

人们对化学毒物的反应和接触的影响有很大的不同 生物毒素。因此，与不同个人接触相关的不良后果的风险 而且种群可能会有很大的差异。基因与环境(G×E)的相互作用可能是显著的 这些风险差异的组成部分。然而，我们在很大程度上仍然对关键的遗传因素和 与特定毒素/毒物的机械联系。因此，我们通过 确定易感个人和人群以进行有效的预防工作仍然十分紧迫 有限的。目前识别G×E相互作用的方法依赖于遗传关联研究，通常 缺乏足够的能力来确定显著的关联，这是由于大量的遗传变异和少量的 接触过的个体的数量。我们建议，以一种根本不同的方法，首先系统地 确定影响对特定毒物/毒素的功能反应的常见人类变种 勾勒出关键候选G、X、E交互作用，以供相关个人和人群有针对性地考虑。 我们将专注于对1490个基因的功能询问，ToxVar集合包含一个聚合频率 -gt；0.1%的功能突变丢失在所有人类群体中进行了评估，以前被确认为 与一种或多种毒物/毒素相互作用。我们争辩说，这些通常在功能上妥协 在相当大比例的人中，基因最有可能影响人类对毒素/毒物的反应。我们 将同时询问这1490个基因中每一个的功能中断对细胞的影响 CRISPR联合筛选与单细胞毒理学相关基因对毒物的反应 表达式目标(ScTRGET)。我们将评估ToxVar-scTRGET方法以从功能上识别 参与细胞对高毒物反应的相关和常见变异基因 在生理上越来越相关的细胞模型中的相关性。

People vary considerably in their response to and in the effects of exposure to chemical toxicants and biological toxins. As a result, the risk of adverse outcomes associated with exposure for different individuals and populations can be widely divergent. Gene by environment (G x E) interactions likely underlie a significant component of these risk differences. However, we remain largely ignorant of both the key genetic factors and the mechanistic association with specific toxins/toxicants. As a result, our capability to mitigate risk by the identification of susceptible individuals and populations to enable effective preventive efforts remain sorely limited. Current approaches to identify G x E interactions rely on genetic association studies which generally lack sufficient power to identify significant associations, due to the large number of genetic variants and small populations of exposed individuals. We propose, in a fundamentally different approach, to first systematically identify the common human variants which impact the functional response to a specific toxicant/toxin to delineate key candidate G X E interactions for targeted consideration in relevant individuals and populations. We will focus on functional interrogation of 1490 genes, the ToxVar set, which contain an aggregate frequency of loss of function mutations of >0.1% in all human populations assessed to date and previously identified as interacting with one or more toxicant/toxins. We contend that these commonly functionally compromised genes are most likely to impact human response to a toxin/toxicant in a significant proportion of people. We will simultaneously query the impact of functional disruption in each of these 1490 genes on the cellular response to a toxicant using coupled CRISPR screening and single cell toxicologically relevant gene expression targets (scTRGETs). We will evaluate the ToxVar-scTRGET approach to identify functionally relevant and commonly variant genes involved in cellular response to selected toxicants of high human relevance in increasingly physiologically relevant cell models.