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

项目总结 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、使人衰弱的疾病 神经发育障碍，总共影响了500多万美国人。尽管有五十多年的历史 在研究方面，没有治愈方法，治疗标准仍然不令人满意。替代性监管失调 人类大脑中的剪接被认为与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，可以在不同的神经元亚型； 然而，这些复杂基因中的大多数的异构体谱系还没有在细胞类型特异的 举止。长阅读测序的最新进展提供了一个前所未有的机会来解决 复杂的可选剪接。因此，为了更好地了解选择性剪接的变化对临床的影响 精神疾病，评估亚型谱系如何影响神经元成熟和突触是至关重要的 以特定于细胞类型的方式起作用。在这里，我们建议对 用单细胞长读测序法研究人类SZ相关基因亚型谱系的复杂性 方法，并研究遗传变异如何影响选择性剪接和影响神经元 成熟和突触功能以亚型特有的方式。该项目将提供全面的 将广泛促进SZ研究的细胞特异性SZ基因全同构体谱系目录。它还将 加深对基因变异和选择性剪接如何影响SZ的理解，有助于更好地 预测临床结果并确定新的治疗干预措施。中使用的单元格方法 该项目一般适用于其他精神障碍的研究，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.