Comparison of HD and HDL2 mouse models to reveal common mechanisms of pathogenesis

HD 和 HDL2 小鼠模型的比较揭示共同的发病机制

• 批准号: 10347570
• 负责人: RUSSELL L MARGOLIS
• 金额: $ 45.03万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-15 至 2024-03-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10347570
• 关键词: Adult; Age-Months; Alleles; Antibodies; Appearance; Autopsy; Bacterial Artificial Chromosomes; Behavior; Biological Assay; Brain; Brain region; Brain-Derived Neurotrophic Factor; CRISPR/Cas technology; Cell model; Cessation of life; Chromosomes; Clinical; Clinical Trials; Cognition Disorders; Corpus striatum structure; Deposition; Development; Disease; Emotional disorder; Exons; Funding; Gene Expression Profile; Genes; Goals; Heterozygote; Human; Huntington Disease; Immune; Inherited; Knock-in; Knock-in Mouse; Lead; Length; Measures; Mitochondria; Modeling; Molecular; Motor; Movement Disorders; Mus; Mutation; Nerve Degeneration; Neurodegenerative Disorders; Neurons; Nuclear Protein; Open Reading Frames; Pathogenesis; Pathogenicity; Pathway interactions; Patients; Peptides; Phenotype; Process; Proteomics; RNA; Research Design; Role; Stains; Synapses; Testing; Therapeutic Intervention; Time; Tissues; Toxic effect; Transcript; Transgenic Organisms; Triplet Multiple Birth; Validation; Withdrawal; brain cell; design; experimental study; falls; gene function; genetic testing; induced pluripotent stem cell; junctophilin; loss of function; motor behavior; motor disorder; mouse model; neuroimaging; neuropathology; neurotoxicity; new therapeutic target; polyglutamine; preclinical development; protein aggregation; therapeutic target; transcriptome; transcriptome sequencing; transcriptomics

Project Summary HD is an autosomal dominant neurodegenerative disease, characterized by movement, cognitive and emotional disorders, with relentless progression to death. There is currently no disease-modifying treatment for HD. Though clinical suppression strategies are in clinical trials or preclinical development, the efficacy and tolerability of these treatments remains unknown, hence there is great need to search, in parallel, for other therapeutic targets. To facilitate prioritizing HD pathways for therapeutic interventions, we have launched an effort to compare HD with Huntington’s disease-like 2 (HDL2). HDL2, discovered and genetically defined by our group, is an adult onset autosomal dominant neurodegenerative disorder that is clinically, genetically, and neuropathologically remarkably similar to HD. Experts cannot distinguish between the two disorders without genetic testing. Both disorders are caused by a repeat expansion mutation; the mutation in HDL2 is caused by a CTG/CAG expansion on chromosome 16q24 in the gene junctophilin-3. Like HD, expanded tracks of polyglutamine probably are the key factor in HDL2 pathogenesis. However, also like HD, RNA transcripts containing the expanded repeat and a loss-of- function the gene in which the repeat is found likely also contribute to pathogenesis. We hypothesize that HD and HDL2 share pathogenic pathways, and that detecting these pathways will lead to further understanding of both disorders and the development of new therapeutic targets. As part of our efforts to test this hypothesis, we propose to use CRISPR/Cas9 and ssDNA donors to generate an HDL2 knock-in (KI) mouse model with either 14 (normal) or ~100 triplets (mutation). We will compare these new mouse lines with the similar KI HD lines Hdh10/+ and HDh111/+. In Aim 1, we will test the hypothesis that the HDL2 and HD expansion mutations results in a similar phenotype by comparing the behavior and neuropathology of HDL2 and HD KI lines. In Aim 2, we will test the hypothesis that similar molecular mechanisms are associated with HD and HDL2 pathogenesis by using RNAseq to compare the pattern of gene expression in the HDL2 and HD model mice. If successful, this project will provide the basis for detailed studies designed to find new therapeutic targets for both HD and HDL2.

项目摘要 HD是一种常染色体显性遗传性神经退行性疾病，其特征是运动、认知和 情绪障碍，并无情地进展到死亡。目前还没有治疗疾病的方法 治疗多发性硬化症。虽然临床抑制策略处于临床试验或临床前开发阶段，但 这些疗法的疗效和耐受性尚不清楚，因此极有必要在 对于其他治疗靶点，也是平行的。为了便于区分用于治疗干预的HD路径的优先顺序， 我们已经发起了一项工作，将HD与亨廷顿病样2(HDL2)进行比较。发现HDL2 根据我们小组的基因定义，是一种成人起病的常染色体显性遗传性神经退行性疾病 这在临床上、基因上和神经病理上都与HD非常相似。专家们无法区分 在没有基因检测的情况下，这两种疾病之间的联系。这两种疾病都是由反复扩张引起的 突变；HDL2的突变是由该基因中染色体16q24上的CTG/CAG扩展引起的 Junctophlin-3。与HD一样，多聚谷氨酰胺的扩展轨迹可能是HDL2的关键因素 发病机制。然而，与HD一样，RNA转录本包含扩展的重复序列和丢失的- 该重复序列所在的基因可能也与发病有关。 我们假设HD和HDL2共享致病途径，检测这些途径将 导致对这两种疾病的进一步了解和新治疗靶点的开发。作为一部分 在检验这一假说的努力中，我们建议使用CRISPR/Cas9和ssDNA捐赠者来生成一个 具有14个(正常)或~100个三胞胎(突变)的HDL2敲入(KI)小鼠模型。我们将对这些进行比较 具有与KI HD相似的Hdh10/+和HDh111/+的新的小鼠品系。在目标1中，我们将检验假设 HDL2和HD扩展突变导致相似的表型通过比较行为和 HDL2和HD KI细胞系的神经病理。在目标2中，我们将检验相似分子的假设 通过使用RNAseq比较HD和HDL2的发病模式，与HD和HDL2的发病机制相关 HDL2和HD模型小鼠的基因表达。如果成功，该项目将为 旨在为HD和HDL2找到新的治疗靶点的详细研究。