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

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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.