Discovery and Functional Studies of Genes for T1D GWAS Susceptibility Loci

描述（由申请人提供）：最近美国和全球1型糖尿病（T1 D）发病率的上升进一步增加了全球医疗服务提供者的经济负担。虽然在了解T1 D的发病机制方面取得了进展，但我们仍然无法预防或逆转这种疾病。通过对人类的大规模遗传学研究，其他人最近发现了人类基因组中约50个不同的染色体区域，可能含有导致T1 D风险或进展的基因。不幸的是，我们仍然不知道这些区域中的哪些特定基因在发挥作用。进一步深入了解T1 D的遗传学将提供更好的风险评估，并可能确定该疾病的治疗靶点。非肥胖型糖尿病（NOD）小鼠与人类T1 D的生理、生化、病理机制以及遗传学特征相似，对T1 D的研究具有重要价值。确定人类中哪些基因实际上起作用的一个有用策略是在NOD小鼠这样的模型中破坏这些基因的功能。然而，目前用于询问大量基因的策略将花费太长时间并且不具有成本效益。在过去的3年里，我们开发了创新和快速的方法来制作转基因动物模型，这种方法具有高度的可重复性以及时间和成本效益。我们是世界上第一个展示靶向锌指核酸酶（ZFN）技术的公司，也是第一个应用TAL效应核酸酶技术的公司之一，靶向和破坏（“敲除”）大鼠中的特定基因，并成功应用这种方法敲除NOD小鼠中的基因。我们还开发了这项技术，能够将非常特定类型的基因突变“敲入”大鼠或小鼠基因组。这是两个非常重要的技术，我们将应用于NOD小鼠模型，以发现和功能研究大量潜在的人类T1 D基因。拟议研究的结果将对我们对T1 D的理解产生巨大而广泛的影响。 公共卫生相关性：开发治疗1型糖尿病（T1 D）的有效疗法的一个重要关键是了解所涉及的基因和这些基因作用的细胞，以靶向新药和新方法。其他研究最近已经产生了关于人类基因组的某些染色体“区域”的重要信息，这些区域可能包含一个或多个有助于T1 D风险和/或进展的基因。然而，我们还不知道这些区域中的许多基因中哪些是重要的。我们将使用一种策略，在自发性T1 D小鼠模型中破坏这些潜在的T1 D疾病基因，并观察破坏的基因对疾病发生率和进展的影响。我们希望发现大量在这种可怕疾病中发挥作用的新基因，这些基因将成为识别高危个体的新生物标志物，并为开发治疗方法提供新的靶点。

DESCRIPTION (provided by applicant): The recent rise of the type 1 diabetes (T1D) incidence both in Americans and globally has further increased the economic burden of the health providers worldwide. While progress has been made to understand the pathogenetic basis of T1D, we are still unable to prevent or reverse the disease. Through large-scale genetics studies in humans, others have recently identified about 50 different chromosome regions in the human genome likely containing genes that contribute to the risk or progression of T1D. Unfortunately, we still do not know for the most part which specific genes within these regions are playing a role. Further insight into the genetics of T1D will provide better risk assessment and potentially identify therapeutic targets of the disease. The Non-Obese Diabetic (NOD) mouse is of tremendous value to T1D research because of its similarity to the humans with the disease, including physiological, biochemical, and disease pathological mechanisms as well as the genetic contribution. A useful strategy to determine which genes in humans are actually playing a role is to disrupt the functions of those genes in a model like the NOD mouse. However, the current strategies for interrogating a large number of genes would take too long and are not cost effective. In the past 3 years, we have developed innovative and rapid ways of making genetically modified animal models which is highly reproducible as well as time and cost efficient. We were the first in the world to demonstrate targeted zinc-finger nuclease (ZFN) technology, and among the first to apply TAL Effector Nuclease technology, to target and disrupt ('knock out'), specific genes in the rat and have successfully applied this approach to knock out a gene in the NOD mouse. We have also developed this technology to be able to 'knock in' very specific types of gene mutations into the rat or mouse genome. These are two very important technologies which we will apply to the NOD mouse model to discover and functionally investigate a large number of the potential human T1D genes. The results of the proposed studies will have a great and broad impact on our understanding of T1D. PUBLIC HEALTH RELEVANCE: One important key to developing effective therapies for the treatment Type 1 Diabetes (T1D) is to understand the genes involved and the cells that these genes work in to target new drugs and approaches. Other studies have recently yielded important information about certain chromosome 'regions' of the human genome which likely contains one or more genes which contribute to T1D risk and/or progression. However, we do not yet know which of the many genes in these regions are important. We will use a strategy where we disrupt these potential T1D disease genes in a mouse model which develops spontaneous T1D and see what the effect of the disrupted gene is on the disease incidence and progression. We expect to uncover a significant number of new genes which are playing a role in this terrible disease and will represent new biomarkers for identifying at risk individuals and provide new targets for therapies to be developed.