High-throughput disease modeling to uncover shared and unique characteristics among neurodegenerative diseases

项目摘要 神经退行性疾病(NDDS)给美国的医疗保健带来了巨大的临床和财政压力 系统。我们目前缺乏FDA批准的有效疗法来阻止或逆转患者的病程 这门课上有很多疾病。通过对NDDS的建模，我们已经开始剖析 与新城疫发病有关的基因和蛋白质。我们发现了核心细胞的微扰 蛋白质折叠和蛋白质周转等过程是许多NDDS的核心。然而，对 管理疾病发展的机制和途径等待着许多新的发展。为了迎接这一挑战， 我们提出了一种新的技术，使用下一代DNA测序方法来检测 在单个实验中建立神经退行性疾病模型，从而增加吞吐量并限制 实验变种。为了捕捉每个NDD模型施加的基本细胞扰动，我们将 描述每个模型对广泛的遗传扰动的反应。随后对这些数据进行分析 将揭示受疾病基因表达影响的细胞途径。 我们将把这个平台应用于肌萎缩侧索硬化症(ALS)和额颞部的研究 痴呆症(FTD)，在临床上发生。与ALS/FTD相关的不同基因突变可以 使患者偏向这一光谱的两端。此外，还有许多遗传变异与 ALS/FTD仍未确定其功能。与ALS/FTD有关的基因已被证明在其中起作用 在许多细胞过程中的作用，包括核糖核酸代谢、核质穿梭和自噬。 成熟。我们的技术平台将允许我们捕捉细胞对数十个基因的反应范围 以及与ALS/FTD的发生有关的等位基因，也确定了人类进一步研究的细胞靶点 神经元。 该项目的目标是：在全基因组范围内利用我们的多路疾病建模平台 用于识别增强或改善ALS/FTD相关基因病理后果的基因的量表 (目标1)，并利用这些发现来验证IPSC皮质神经元的潜在治疗线索(目标2)。

Project Summary Neurodegenerative diseases (NDDs) present a large clinical and financial strain on the US healthcare system. We currently lack effective FDA approved therapeutics that halt or reverse the course of disease for many diseases in this class. Through modeling NDDs, we have begun to dissect the pathological impact of genes and proteins implicated in NDD development. We have discovered perturbations of core cellular processes such as protein folding and protein turnover are central to many NDDs. However, understandings of mechanisms and pathways governing disease development awaits for many NDDs. To approach this challenge, we propose a novel technology that using next generation DNA sequencing methods to examine multiple neurodegenerative disease models within a single experiment, thereby increasing throughput and limiting inter- experimental variation. To capture fundamental cellular perturbations imposed by each NDD model, we will characterize each model’s response to a wide range of genetic perturbations. Subsequent analysis of these data will reveal cellular pathways impacted by disease gene expression. We will apply this platform towards the study of Amyotrophic Lateral Sclerosis (ALS) and Frontotemporal Dementia (FTD), which occur on a clinical spectrum. Mutations in different genes implicated in ALS/FTD can bias patients towards either end of this spectrum. Additionally, there are many genetic variants implicated in ALS/FTD which remain functionally uncharacterized. The genes implicated in ALS/FTD have been shown to play a role in many cellular processes, including RNA metabolism, nucleocytoplasmic shuttling, and autophagosome maturation. Our technological platform will allow us to capture the scope of cellular responses to dozens of genes and alleles implicated in the development of ALS/FTD and also identify cellular targets for further study in human neurons. The goals of this project are to: leverage our multiplexed disease modeling platform on a genome-wide scale to identify of genes that enhance or ameliorate pathological consequences of genes implicated in ALS/FTD (Aim 1), and to harness these findings to validate potential therapeutic leads in iPSC cortical neurons (Aim 2).