rAAV-mediated knockdown for Huntington's disease

rAAV 介导的亨廷顿舞蹈病敲除

基本信息

批准号：
7350216
负责人：
RONALD J MANDEL
金额：
$ 28.53万
依托单位：
UNIVERSITY OF FLORIDA
依托单位国家：
美国
项目类别：
财政年份：
2006
资助国家：
美国
起止时间：
2006-02-01 至 2011-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7350216
关键词：
Affect Alleles Area Basal Ganglia Behavior Birth Brain CAG repeat Cell Death Cells Cessation of life Cognitive Cognitive deficits Complement Corpus striatum structure Data Defect Development Disease Disease Progression Disease model Engineering Exons Functional disorder GAG Gene Gene Transfer Genes Genetic Human Huntington Disease Inclusion Bodies Induced Mutation Inherited Knock-in Mouse Knowledge Lead Link Mediating Messenger RNA Metric Modeling Motor Mus Neurodegenerative Disorders Neurons Nuclear Inclusion Nuclear Protein Nuclear Proteins Numbers Pathology Pathway interactions Patients Proteins Recombinants Research Personnel Series Small Interfering RNA Specificity Symptoms Testing Time Toxic effect Transcript Transgenic Mice Transgenic Organisms Work adeno-associated viral vector base design disease phenotype gain of function human Huntingtin protein knock-down mouse model mutant nervous system disorder neuron loss neuropathology postnatal prevent programs protein aggregation putamen success

项目摘要

Huntington's disease (HD) is an autosomal dominant inherited progressive neurological disorder. The neuorpathological symptoms appear to be caused by progressive dysfunction and death of neostriatal neurons and/or the cortico-striatal tract. The HD gene was first described in 1993 and the gene defect was identified as an expansion of a series of CAG repeats above a threshold level in exon 1 of a gene encoding a protein called huntingtin. Subsequently, similar repeat expansions have been identified in eight other genes that are known to cause different neurological disorders when the trinucleotide expansions reach thresholds typical for each disease. These different disorders are also conceptually linked because, like HD, symptoms are caused by the dysfunction of specific neurons in the CNS. Moreover, in all CAG expansion mutation- induced disorders, the mutant protein seems to induce abnormal nuclear protein aggregations termed neuronal intranuclear inclusions. In HD, the best data seem to indicate that both a toxic gain of function and a loss of some normal function in the mutant form of huntingtin leads to neuropathology. Thus, our proposal aims to use recombinant adeno-associated viral vectors to deliver genes encoding constructs to knock-down huntingtin in striatal neurons. Long-term knock-down of striatal huntingtin will test the hypothesis that reduced huntingtin expression should limit the formation of neuronal Intranuclear inclusions in transduced neurons and thereby slow disease progression in mouse models of HD. We will use small interfering RNAs (siRNAs) to knock-down huntingtin in striatal neurons. Moreover, we propose two ways to test the hypothesis that normal huntingtin expression is beneficial.. First, we will compare the effects of huntingtin knock-down in both knock-in HD mice and transgenic HD mice. Since transgenic HD mice still express the normal amount of mouse huntingtin, our hypothesis predicts that huntingtin knock-down will be more beneficial in the transgenic mice compared to the knock-in HD mice that have only mutant huntingtin expression. Second, it is unclear whether striatal dysfunction originates in a cell autonomous fashion in striatal neurons or is caused by mutant huntingtin-induced cortico-striatal dysfunction. We will directly test this hypothesis by transducing cortical areas vs. striatal areas. We will use reduction in the amount of striatal neuronal nuclear inclusions and reversal of known transcriptional changes as our initial screen for beneficial effects followed by detailed functional analysis of motor behavior as metric of success of these strategies.

亨廷顿病（HD）是一种常染色体显性遗传的进行性神经系统疾病。这神经病理学症状似乎是由新纹状体进行性功能障碍和死亡引起的神经元和/或皮质纹状体束。 HD基因于1993年首次被描述，该基因缺陷为被鉴定为编码 a 的基因的外显子 1 中一系列高于阈值水平的 CAG 重复的扩展。称为亨廷顿蛋白的蛋白质。随后，在其他八个基因中也发现了类似的重复扩展当三核苷酸扩增达到阈值时，已知会导致不同的神经系统疾病每种疾病的典型特征。这些不同的疾病在概念上也有联系，因为像 HD 一样，症状是由中枢神经系统中特定神经元功能障碍引起的。而且，在所有CAG扩张突变中—— 引起的疾病，突变蛋白似乎会诱导异常的核蛋白聚集，称为神经元核内包涵体。在 HD 中，最好的数据似乎表明，功能的毒性增益和亨廷顿蛋白突变体中某些正常功能的丧失会导致神经病理学。因此，我们的建议旨在使用重组腺相关病毒载体将编码构建体的基因传递至敲除纹状体神经元中的亨廷顿蛋白。纹状体亨廷顿蛋白的长期敲除将检验以下假设：亨廷顿蛋白表达的减少应限制转导中神经元核内包涵体的形成神经元，从而减缓 HD 小鼠模型的疾病进展。我们将使用小干扰 RNA (siRNA) 敲低纹状体神经元中的亨廷顿蛋白。此外，我们提出了两种方法来测试假设正常的亨廷顿蛋白表达是有益的。首先，我们将比较亨廷顿蛋白的效果敲入 HD 小鼠和转基因 HD 小鼠中均出现敲低。由于转基因 HD 小鼠仍然表达正常量的小鼠亨廷顿蛋白，我们的假设预测亨廷顿蛋白敲低会更多与仅具有突变亨廷顿蛋白的敲入 HD 小鼠相比，转基因小鼠有益表达。其次，尚不清楚纹状体功能障碍是否起源于细胞自主方式。纹状体神经元或由突变亨廷顿蛋白引起的皮质纹状体功能障碍引起。我们直接测试这一假设是通过转换皮质区域与纹状体区域来实现的。我们将减少纹状体的数量神经元核内含物和已知转录变化的逆转作为我们有益的初步筛选效果，然后对运动行为进行详细的功能分析，作为这些策略成功的衡量标准。