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Genetic Suppression of SMN Mutations in Spinal Muscular Atrophy

脊髓性肌萎缩症中 SMN 突变的基因抑制

基本信息

批准号：
10430238
负责人：
ARTHUR H. M. BURGHES
金额：
$ 51.65万
依托单位：
OHIO STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-07-01 至 2026-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10430238
关键词：
Address Affect Alleles Animals Axon Binding Biochemical Biochemical Pathway Biological Assay Birth C-terminal Cell Culture Techniques Cell Line Cell Survival Cells Clustered Regularly Interspaced Short Palindromic Repeats Complement Critical Pathways DNA Sequence Alteration Defect Development Disease Electrophysiology (science)Engineering Excision Exons Genes Genetic Genetic Suppression Human Lead Length Missense Mutation Modeling Motor Mus Mutate Mutation N-terminal Neuromuscular Diseases Pathogenesis Pathway interactions Patients Phenotype Plant Roots Proteins RNA RNA Splicing Reporting Role SMN deficiency SMN protein (spinal muscular atrophy)SMN1 gene SMN2 gene Small Nuclear Ribonucleoproteins Spinal Muscular Atrophy Study models System Tertiary Protein Structure Testing Transgenic Mice Translating Translations U7 Small Nuclear Ribonucleoprotein Zebrafish effective therapy loss of function motor neuron function mouse model mutant nervous system disorder overexpression profilin snRNP Biogenesis survival motor neuron gene

项目摘要

Spinal Muscular Atrophy is a devastating neuromuscular disease caused by insufficient amounts of SMN protein. SMA is caused by loss or mutation of the SMN1 gene and retention of the SMN2 gene. The SMN2 gene is a modifier of phenotype where milder SMA cases having more copies of SMN2. Rarely SMA patients have a missense mutation in the SMN1 gene. We can use these mutations and the protein domains they disrupt to study the function of the SMN protein. We have shown that SMA missense mutations are not functional by themselves but can function in the presence of some full-length wild-type SMN protein. Furthermore, we have shown that N and C-terminal SMN missense mutations can complement each other and rescue snRNP assembly in the complete absence of full-length wild-type SMN protein in mice. We have developed cell line that conditionally removes functional SMN to allow us test SMN missense mutations in culture. We have also used this cell line to test for suppressors of the SMNE134K mutation. We have identified a suppressor in the SmF protein that fully restores snRNP assembly lost due to the SMN E134K mutation. We now have a system to screen for suppressors of SMN missense mutations. In this proposal we will test the SmF suppressor we have found in two different SMN E134K mouse models and determine if this mutation rescues the SMA phenotype and survival of the SMA mouse. Thus, we can study the separate functions of SMN in snRNP assembly from the function of SMN in the axon. We will screen for additional suppressors using other SMN patient derived mutations to test other functional domains of SMN. We will investigate the role of SMN in the axon independent of Sm assembly by introducing HuD and truncated forms of SMN into the SMA mice via scAAV9. We have shown in cells that Smn exon2B is not required for cell survival. We will test scAAV9-Smn∆2 in SMA mice to confirm this finding and rescue the SMA phenotype. Finally we will test the role of profilin in axonal function in the SMA mouse using the SMNS230L mutation. Using genetic mutations we can dissect the functions of SMN in splicing and in the axon to resolve the underlying mechanism by which reduced SMN protein causes SMA.

脊髓性肌萎缩症是由SMN蛋白不足引起的一种毁灭性的神经肌肉疾病。