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

• 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
• 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' 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

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.

项目摘要 突触传递和可塑性是神经元功能和突触失调的基础 蛋白质表达是神经发育障碍（例如自闭症）的直接原因。超过一百de Novo Syngap1中的功能丧失突变与自闭症光谱无关紧要 疾病和智力残疾。剪接切换寡核苷酸（SSO）的最新成功表明 通过遗传和SSO介导的消融重定向剪接是一种有前途的营救方法 单倍不足。我们已经确定了Syngap1中的替代剪接事件，这导致了胡说八道 - 小鼠和人类发育过程中介导的mRNA衰变（NMD）。确定syngap1是否 NMD外显子是一个可行的治疗靶标，我们使用遗传研究了调节机制及其功能 方法，并确定Syngap1 NMD外显子的遗传缺失和SSO抑制是否可以 挽救小鼠突变体和IPPC衍生神经元中的杂合敲除表型。之上 完成，该项目将提供有关此Syngap1 NMD外显子的生理功能的遗传见解 并产生关键的临床前试剂，以恢复单倍不足的Syngap1蛋白表达。