Rescuing SYNGAP1 haploinsufficiency by redirecting alternative splicing

通过重定向选择性剪接挽救 SYNGAP1 单倍体不足

基本信息

批准号：
10660668
负责人：
Xiaochang Zhang
金额：
$ 64.41万
依托单位：
UNIVERSITY OF CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-04-01 至 2028-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10660668
关键词：
3&apos Splice Site AMPA Receptors Ablation Alleles Alternative Splicing Animals Behavior Behavioral Brain Cells Coupled Defect Development Disease Electrophysiology (science)Elements Event Exons Genetic Genetically Engineered Mouse Glutamates Heterozygote Hippocampus Human Human Development Impairment Induced pluripotent stem cell derived neurons Intellectual functioning disability Knock-out Knockout Mice Lead Long-Term Potentiation Loss of Heterozygosity Mediating Molecular Mus Mutant Strains Mice Neocortex Neurodevelopmental Disorder Neurons Oligonucleotides Patients Phenotype Physiological Pilot Projects Protein Isoforms Proteins RNA Splicing Reagent Reporting Research Project Grants SYNGAP1 Synapses Synaptic Transmission Synaptic plasticity Testing Toxic effect Vertebral column autism spectrum disorder efficacy evaluation genetic approach insight learned behavior loss of function loss of function mutation mRNA Decay mouse development mouse genetics mouse model neocortical neurodevelopment postnatal pre-clinical protein expression success therapeutic target

项目摘要

PROJECT SUMMARY Synaptic transmission and plasticity are fundamental to neuronal functions, and dysregulations of synaptic protein expression are direct causes of neurodevelopmental disorders such as autism. Over one hundred de novo loss-of-function mutations in SYNGAP1 have been unambiguously associated with autism spectral disorders and intellectual disability. Recent success in splice-switching oligonucleotides (SSOs) suggests that redirecting splicing through genetic and SSO-mediated ablations is a promising approach to rescue haploinsufficiency. We have identified an alternative splicing event in SYNGAP1 that leads to nonsense- mediated mRNA decay (NMD) during mouse and human development. To determine whether the SYNGAP1 NMD exon is a viable therapeutic target, we investigate the regulatory mechanism and its functions using genetic approaches, and determine whether genetic deletion and SSO suppression of the SYNGAP1 NMD exon can rescue heterozygous knockout phenotypes in mouse mutants and patient-iPSC-derived neurons. Upon completion, this project will provide genetic insights into the physiological functions of this SYNGAP1 NMD exon and generate critical preclinical reagents to restore SYNGAP1 protein expression from haploinsufficiency.

项目概要突触传递和可塑性是神经元功能的基础，突触的失调蛋白质表达是自闭症等神经发育障碍的直接原因。一百多德 SYNGAP1 的 novo 功能丧失突变与自闭症谱系明确相关障碍和智力障碍。最近剪接转换寡核苷酸（SSO）的成功表明通过遗传和 SSO 介导的消融重定向剪接是一种有前途的拯救方法单倍体不足。我们在 SYNGAP1 中发现了一个替代剪接事件，它会导致无意义的结果—— 小鼠和人类发育过程中介导的 mRNA 衰减（NMD）。判断SYNGAP1是否 NMD 外显子是一个可行的治疗靶点，我们利用遗传学研究其调控机制及其功能方法，并确定 SYNGAP1 NMD 外显子的基因删除和 SSO 抑制是否可以拯救小鼠突变体和患者 iPSC 衍生神经元中的杂合敲除表型。之上完成后，该项目将为 SYNGAP1 NMD 外显子的生理功能提供遗传学见解并生成关键的临床前试剂以恢复单倍体不足的 SYNGAP1 蛋白表达。