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

项目摘要/摘要 阿尔茨海默病(AD)是老年人中最常见的痴呆症，由复杂的 遗传和环境风险因素的相互作用。预计到2050年，将有1.3亿人患上阿尔茨海默病， 构成了临床上迫切需要创造有效的治疗方法和疗法。以前的治疗方法有 专注于解决病理特征，如老年斑(Ab)和神经原纤维缠结(Tau)，但 未能改善患者的认知功能。新兴的基于RNA测序的方法已经 研究表明，RNA加工中断事件与认知功能下降高度相关，并可以 作为疾病的生物标志物。然而，这些技术还不能捕捉到RNA的加工 阿尔茨海默病的时空变化对像大脑这样复杂的组织至关重要。在此应用程序中， 我们建议开发一种新的mRNA测序方法并应用于AD患者组织，以揭示 首次在单细胞水平上处理复杂的RNA模式。我们将利用已有的资源 建立和优化我们的技术的实验室模型(目标1)，然后对皮质(额叶、枕叶、 AD患者和对照组的颞叶和顶叶)、海马体和脑干组织(目标2)。这些 研究结果将以前所未有的分辨率揭示信使核糖核酸多聚腺苷化位点使用的复杂性。此外 对于AD，我们的研究将广泛影响所有组织中mRNA3‘端多样性的研究，因为它将适用于 任何冰冻或新鲜的组织来源。

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.