Resolving complex alternative splicing of psychiatric disease genes using single-cell approaches

项目摘要 精神分裂症（SZ）、双相情感障碍（BD）和自闭症谱系障碍（ASD）是常见的和衰弱的 这些神经发育障碍共同影响了超过500万美国人。尽管五十多年来 尽管研究进展缓慢，但没有治愈的方法，治疗标准仍然不能令人满意。替代调节失调 人类大脑中的剪接与SZ有关。在具有关键大脑功能的基因中，许多基因具有 大量的外显子，导致复杂的剪接模式在神经元亚型之间可能有所不同; 然而，大多数这些复杂基因的同种型库还没有在细胞类型特异性的 方式长读段测序的最新进展为解决这一问题提供了前所未有的机会。 复杂可变剪接因此，为了更好地了解选择性剪接的变化对临床的影响， 在精神疾病中，评估同种型库如何影响神经元成熟和突触形成是至关重要的。 以细胞类型特异性方式起作用。在这里，我们建议目录和功能特性， 使用单细胞长读段测序的SZ相关基因的人类同种型库的复杂性 的方法，并研究遗传变异如何影响选择性剪接和影响神经元 成熟和突触功能的亚型特异性的方式。该项目将提供全面的 SZ基因的细胞特异性完全同种型库的目录，将广泛促进SZ研究。它还将 加深对遗传变异和选择性剪接如何导致SZ的理解， 预测临床结果并确定新的治疗干预措施。本研究中使用的单细胞方法 项目一般适用于研究其他精神障碍，如BD和ASD。

PROJECT SUMMARY Schizophrenia (SZ), bipolar disorder (BD) and autism spectrum disorder (ASD) are common and debilitating neurodevelopmental disorders that together affect more than 5 million Americans. Despite more than fifty years of research, no cures exist and the standard of treatment remains unsatisfactory. Dysregulation of alternative splicing in the human brain has been implicated in SZ. Among genes with critical brain functions, many have a large number of exons, resulting in complex splicing patterns that can vary between neuronal subtypes; however, the isoform repertoires of most of these complex genes have not been resolved in a cell-type specific manner. Recent advances in long read sequencing have provided an unprecedented opportunity to resolve complex alternative splicing. Thus, to better understand the clinical impact of changes of alternative splicing in psychiatric diseases, it is critical to evaluate how isoform repertoire impact neuronal maturation and synaptic function in a cell-type specific manner. Here, we propose to catalog and functionally characterize the complexity of human isoform repertoires of SZ-associated genes using a single-cell long-read sequencing based approach, and to study how genetic variations influence alternative splicing and impact neuronal maturation and synaptic function in a subtype specific manner. This project will provide comprehensive catalogs of cell-specific full isoform repertoires of SZ genes that will broadly facilitate SZ research. It will also deepen the understanding of how genetic variations and alternative splicing contribute to SZ, help better predict the clinical outcome and identify novel therapeutic interventions. The single cell approach used in this project is generally applicable to study other mental disorders such as BD and ASD.