Experimental Cellular Approaches to Genotype × Environment Interaction

项目总结 我们建议在细胞流行病学领域进行一项新的研究，这是通过 诱导多能干细胞(IPSC)技术的最新革命。众所周知，有许多细胞-- 由于必须依赖于容易获得的限制而错过的特定功能和行为 细胞，如血细胞或淋巴母细胞系，用于疾病病因、风险的流行病学研究 因素和生物标记物鉴定。IPSC技术的进步现在允许我们考虑非侵入性 对人类受试者适合疾病的细胞类型进行大规模深层细胞表型鉴定。健壮 从血细胞中衍生出IPSC系并将其分化为器官特异性细胞类型是可能的。一个 IPSC来源的细胞的重要好处是，观察到的生物变异主要代表遗传 影响，因为历史有机体环境的大部分表观遗传记忆已经丢失。 我们建议的研究涉及一种创新的人类基因×环境实验方法 互动(GEI)。虽然GEI被认为广泛存在，但在人类中的研究相对较少，原因是 环境异质性和控制环境暴露的难度。我们基于iPSC的移动电话 方法允许我们通过检查细胞表型变异来严格地通过实验测试GEI 在一次受控的环境挑战之后。我们的研究将是第一个也是最大规模的模拟人类GEI的研究 在两种不同的IPSC来源的细胞类型中。首先，我们将确定蛇毒的预期神经毒性作用 在神经干细胞(NSCs)中是由基因驱动的，其次，可疑的差异反应是否 肺泡上皮2型细胞(AT2s)对环境污染物的暴露有一定的遗传学基础。 该项目将利用现有的一项重要人力资源--墨西哥裔美国人家庭研究(MAFS)。我们 将使用来自400名MAFS参与者的现有冷冻保存的IPSC品系来生成具有良好特性的 NSCs和AT2。我们提出了一种新的实验和有效的基于系谱的方法来研究遗传 细胞对环境应激反应的基础(即，GEI)，这在以前很难评估。我们的 目的是：1)评估神经干细胞对蛇毒反应的遗传基础；2)评估AT2反应的遗传基础 苯并[a]芘污染；3)检测环境干扰细胞转录的遗传基础 一致性/动态平衡；以及4)确定细胞应激恢复能力对人体器官的多效性效应 与健康相关的表型。 该项目将采用一种基于谱系的新的实验和有效的方法来研究人类GEI， 此前很难对其进行评估。也有助于建立流行病学标尺的可行性。 利用IPSC技术解决生物医学问题。最后，我们预计拟议的项目将 严格建立影响与人类健康相关的复杂表型的GEI的细胞学基础。

PROJECT SUMMARY We propose to perform a novel study in the field of cellular epidemiology, that has been made possible by the recent revolution in induced pluripotent stem cell (iPSC) technology. It is well known that there are many cell- specific functions and behaviors that have been missed by the limitations of having to rely on easily obtainable cells, such as blood cells or lymphoblastoid cell lines, for epidemiological studies of disease causation, risk factors, and biomarker identification. Advances in iPSC technologies now allow us to consider non-invasive large-scale deep cellular phenotyping efforts on disease-appropriate cell types in human subjects. Robust derivation of iPSC lines and their differentiation into organ-specific cell types is possible from blood cells. An important benefit of iPSC-derived cells is that observed biological variation primarily represents genetic influences, since most of the epigenetic memory of the historical organismal environment is lost. Our proposed study involves an innovative experimental approach to human genotype×environment interaction (GEI). While GEI is thought to exist widely, it is relatively poorly studied in humans due to environmental heterogeneity and the difficulty of controlling environmental exposures. Our iPSC-based cellular approach allows us to rigorously test for GEI experimentally by examining cellular phenotypic variation before and after a controlled environmental challenge. Our study will be the first and largest study to model human GEI in two different iPSC-derived cell types. First, we will determine if the expected neurotoxic effect of snake venom in neural stem cells (NSCs) is genetically driven, and secondly, whether the suspected differential response of alveolar epithelial type 2 cells (AT2s) to environmental pollutant exposure has a genetic basis. This project will leverage a major existing human resource, the Mexican American Family Study (MAFS). We will use existing cryo-preserved iPSC lines from 400 MAFS participants for the generation of well-characterized NSCs and AT2. We propose a novel experimental and efficient pedigree-based approach for studying the genetic basis of cellular response to environmental stress (i.e, GEI), which has previously been difficult to assess. Our aims are: 1) assess genetic basis of NSC response to snake venom; 2) assess genetic basis of AT2 response to a benzo[a]pyrene pollution; 3) examine the genetic basis of environmental disruption of cellular transcriptional coherence/homeostasis; and 4) identify pleiotropic effects of cellular stress resilience on human organismal phenotypes relevant to health. This project will employ a novel experimental and efficient pedigree-based approach for studying human GEI, which has previously been difficult to assess. It also will help establish the feasibility of epidemiological scale utilization of iPSC technology to attack biomedical problems. Finally, we expect that the proposed project will rigorously establish the cellular basis of GEI influencing complex phenotypes of relevance to human health.