Multilineage DAmFRET to investigate AD/ADRD protein phase behavior in neural tissue models

项目摘要/摘要 阿尔茨海默病和相关痴呆(AD/ADRD)是最常见的痴呆症形式。不是 有治疗方法可以阻止或预防它们。AD/ADRD在一定程度上是由一个或更多的集合引起的 一种特定蛋白质的组合。TDP-43是为数不多的此类蛋白质之一。它的特征是转变为 在很大一部分痴呆症病例中，从动态组装的液体阶段到僵硬的致病阶段。 这种转变是在不同细胞之间相互作用的渐进式变化的背景下发生的 大脑中的类型。这些分子和细胞变化的相互依赖可能决定了临床 AD/ADRD的课程，但要理解这种相互依赖，我们需要新的工具来进行比较 脑细胞相互作用背景下的蛋白质相变及其表型效应 类型。我们在这里建议通过修改基于细胞的分布式生物物理方法来开发这样的工具 用于人诱导多能干细胞(HiPSC)来源脑组织的两氟FRET(DAmFRET) 模型(目标1)。我们将同时使用DAmFRET开发TDP-43作为细胞类型特异性的报告 蛋白质相变的动态平衡差异(目标2)。通过实现这些目标，我们将拥有 创建并验证了一种独特的强大工具，用于更好地了解复杂的分子和细胞 AD/ADRD的原因。

Project Summary/Abstract Alzheimer's Disease and Related Dementias (AD/ADRD) are the most common forms of dementia. No treatments exist to stop or prevent them. AD/ADRD are caused in part by the aggregation of one, or more often a combination of, specific proteins. TDP-43 is one of a handful of such proteins. It characteristically transitions from a dynamic liquid phase of assembly to a rigid and pathogenic phase in a large fraction of dementia cases. This transition occurs against a backdrop of progressive changes in the interactions between different cell types in the brain. An interdependence of these molecular and cellular changes likely determines the clinical courses of AD/ADRD, but to understand that interdependence, we need new tools with which to compare protein phase transitions and their corresponding phenotype effects in the context of interacting brain cell types. We propose here to develop such a tool by modifying a cell-based biophysical method, Distributed Amphifluoric FRET (DAmFRET) for use in human induced pluripotent stem cell (hiPSC)-derived brain tissue models (Aim 1). We will simultaneously use DAmFRET to develop TDP-43 as a reporter of cell type-specific differences in the homeostasis of protein phase transitions (Aim 2). By completing these aims, we will have created and validated a uniquely powerful tool for better understanding the complex molecular and cellular causes of AD/ADRD.