Developing single nuclear polyAClick-sequencing to profile mRNA 3'-end diversity at the single cell level in Alzheimer's disease.

项目总结/摘要 阿尔茨海默氏病（AD）是老年人中最普遍的痴呆形式，并且由复杂的神经系统疾病引起。 遗传和环境风险因素的相互作用。估计到2050年将有1.3亿人患AD， 构成了对有效治疗和疗法的迫切临床需求。既往治疗有 专注于解决病理标志，如老年斑（Ab）和神经元缠结（tau），但 并不能改善患者的认知功能。新兴的基于RNA测序的方法 研究表明，RNA加工过程的中断与认知功能的下降高度相关， 作为疾病的生物标志物。然而，这些技术未能捕捉到RNA加工过程， 在AD中以时空的方式变化，这在像大脑这样复杂的组织中是至关重要的。在本申请中， 我们建议开发一种新的mRNA测序方法，并将其应用于AD患者组织，以揭示 这是第一次在单细胞水平上研究复杂的RNA加工模式。我们将利用现有的 实验室模型，以建立和优化我们的技术（目标1）之前，剖析皮层（额叶，枕叶， 颞叶和顶叶）、海马和来自AD患者和对照的脑干组织（目的2）。这些 研究结果将揭示mRNA多聚腺苷酸化位点使用的复杂性，在前所未有的分辨率。此外 我们的研究将广泛影响所有组织中mRNA 3 '端多样性的研究，因为它将适用于 任何冷冻或新鲜组织来源。

Project Summary/Abstract Alzheimer’s disease (AD) is the most prevalent form of dementia in the elderly and is caused by complex interactions of genetic and environmental risk factors. An estimated 130 million people will develop AD by 2050, constituting an urgent clinical need for effective treatments and therapies to be created. Prior therapies have focused on resolving pathological hallmarks, such as senile plaques (Ab) and neurofibrillary tangles (tau), but have failed to improve cognitive function in patients. Emerging RNA-sequencing-based approaches have revealed that disrupted RNA processing events are highly correlated with declining cognitive function and can serve as disease biomarkers. However, these technologies have fallen short of capturing the RNA processing changes in AD in a spatiotemporal manner that is critical in tissue as complex as the brain. In this application, we propose to develop and apply a novel mRNA sequencing method to AD patient tissues to reveal complex RNA processing patterns at the single cell level for the first time. We will utilize preexisting laboratory models to establish and optimize our technology (Aim 1) before profiling the cortex (frontal, occipital, temporal, and parietal), hippocampus, and brain stem tissue from patients with AD and control (Aim 2). These findings will reveal the complexity of mRNA polyadenylation site usage at unprecedented resolution. In addition to AD, our research will broadly impact the study of mRNA 3’-end diversity across all tissues, as it will apply to any frozen or fresh tissue source.