Defining the Genetic Architecture of the Glutathione Redox System

项目摘要/摘要 内源性抗氧化剂谷胱甘肽(GSH)赋予细胞抵抗压力，维持生存， 并适当调节基本信号通路。组织内的GSH水平，以及 其还原和氧化形式的比例似乎是与生俱来的，表明真核生物继承了 合成和代谢GSH的相对能力。尽管有这种明显的遗传效应，但基因 对谷胱甘肽系统的调控仍不明确。相反，目前的知识仅限于一小部分 谷胱甘肽还原酶(Gr)和谷胱甘肽过氧化物酶-1(GPX-1)等典型GSH基因的数量。 我们的初步研究表明，这一系统的遗传调控实际上更为复杂，可能 涉及一组新的基因。这些初步努力是基于有时受到限制的硅胶方法。 因此，现在最重要的是进行高精度的基因定位，以验证我们新发现的基因座， 并确定以前被忽视的基因座。在当前项目中，我们将通过以下方式完成这些关键任务 验证我们的中心假设：谷胱甘肽系统受遗传变异的调节：1)典型的谷胱甘肽 基因，包括Gr和gpx-1；ii)新基因，如与RAR相关的孤儿受体α(RoRα)，其 函数是基本GSH系统的外部函数，我们预计其数量将超过正则函数 谷胱甘肽基因。我们将通过一种策略来检验这一假设，该策略将正向遗传学方法与 创新多样性繁殖(DO)小鼠种群，它模拟在人类中发现的遗传多样性，以及一个 基于CRISPR/Cas9技术创建的新型小鼠模型的反向遗传学方法。我们会 解决以下具体目标：1)量化核心GSH表型在遗传上的遗传力 多样性群体；2)定义与GSH系统相关的基因组区域，并描绘共享和 组织特异性基因座；以及3)优先选择候选基因，并开始对最引人注目的基因进行功能分析 候选人。这些研究将定义一种不可或缺的生化物质的基本遗传结构 控制细胞抗应激和生存的系统。从这些努力中获得的知识将为 未来旨在了解GSH基因对细胞的影响的一系列临床和机制研究 这些数据将为保持组织完整性的创新疗法奠定基础 为患有退行性疾病的患者提供医疗服务，从而延长他们的健康寿命，改善他们的生活质量。

PROJECT SUMMARY/ABSTRACT The endogenous antioxidant glutathione (GSH) confers cells with the ability to resist stress, maintain survival, and properly regulate fundamental signaling pathways. The levels of GSH within a tissue, as well as the proportion of its reduced and oxidized forms, appear to be innate, demonstrating that eukaryotes inherit the relative capacity to synthesize and metabolize GSH. Despite this apparent genetic effect, the genetic regulation of the GSH system remains poorly defined. Instead, knowledge is currently limited to a small number of canonical GSH genes such as glutathione reductase (Gr) and glutathione peroxidase-1 (Gpx-1). Our preliminary studies revealed that the genetic regulation of this system is actually more complex and may involve a novel set of genes. Those preliminary efforts were based on in silico methods that are at times limited in power, so it is now paramount to perform high precision gene mapping to validate our newly discovered loci, and to identify previously overlooked loci. In the current project, we will accomplish those crucial tasks by testing our central hypothesis: that the GSH system is regulated by genetic variation within i) canonical GSH genes, including Gr and Gpx-1, and ii) novel genes, such as the RAR-related orphan receptor α (Rorα), whose functions are external to the basic GSH system, and whose number we expect to exceed that of canonical GSH genes. We will test the hypothesis with a strategy that couples a forward genetics approach with the innovative Diversity Outbred (DO) mouse stock, which models the genetic diversity found in humans, and a reverse genetics approach based on novel mouse models created with CRISPR/Cas9 technology. We will address the following specific aims: 1) to quantify the heritability of core GSH phenotypes in a genetically diverse population; 2) to define genomic regions associated with the GSH system, and delineate shared and tissue-specific loci; and 3) to prioritize candidate genes, and initiate functional analyses of the most compelling candidates. These studies will define the fundamental genetic architecture of an indispensable biochemical system that governs cellular stress resistance and survival. Knowledge gained from these efforts will inform a series of future clinical and mechanistic studies aimed at understanding the impact of GSH genes on cellular damage during stress, and the data will build a foundation for innovative therapies to maintain tissue integrity in patients with degenerative diseases, thereby increasing their health spans and improving their qualities of life.