Antisense Oligonucleotides for Targeting Nonsense-mediated mRNA Decay in Frontotemporal Dementia

反义寡核苷酸用于靶向额颞叶痴呆中无义介导的 mRNA 衰减

• 批准号: 10621865
• 负责人: Andrew D. Nguyen
• 金额: $ 37.88万
• 依托单位: SAINT LOUIS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-01 至 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10621865
• 关键词: Address; Affect; Antibodies; Antisense Oligonucleotides; Behavioral; Binding; Biological Markers; Brain; C-terminal; Cathepsins; Cell Separation; Cell model; Cells; Clinical Trials; Collaborations; Complex; Cysteine; Data; Diagnosis; Disease; Duchenne muscular dystrophy; Exons; Fibroblasts; Frameshift Mutation; Frontotemporal Dementia; Gliosis; Goals; Human; Huntington Disease; Individual; Induced pluripotent stem cell derived neurons; Injections; Intranasal Administration; Knock-in Mouse; Light; Mediating; Messenger RNA; Methods; Modeling; Molecular; Mus; Mutation; Nerve Degeneration; Neurodegenerative Disorders; Nonsense Codon; Nonsense Mutation; Obsessive compulsive behavior; PGRN gene; Pathology; Pathway interactions; Patients; Peptide Hydrolases; Pharmacologic Substance; Phenotype; Phosphorylation; Protein Secretion; Protein Truncation; Proteins; Reporter; Social Dominance; Spinal Muscular Atrophy; Terminator Codon; Testing; Therapeutic; behavior test; causal variant; comparative efficacy; design; granulin; improved; in vivo evaluation; insight; mRNA Decay; mouse model; mutant; nervous system disorder; neurofilament; neuropathology; novel therapeutic intervention; pre-clinical; predictive modeling; premature; prevent; protein TDP-43; tool

Project Summary Progranulin is a lysosomal and secreted protein that contains multiple cysteine-rich granulin domains; its precise molecular function remains unknown. Progranulin (GRN) mutations are causal for frontotemporal dementia (FTD), which is a devastating disease with a mean survival of 3.8 years from diagnosis and no cure currently available. Since progranulin-deficient FTD is a disease of haploinsufficiency, strategies aimed at increasing progranulin levels are feasible therapeutic approaches. We recently generated a knock-in mouse model of FTD harboring the common patient nonsense mutation GRNR493X, and we established that the nonsense-mediated mRNA decay (NMD) pathway contributes to the markedly reduced mutant progranulin mRNA levels in this mouse model, as wel`l as in patient-derived fibroblasts containing the GRNR493X mutation. Moreover, our cell-based studies indicate that the progranulin R493X mutant protein is functional when re- expressed in progranulin-deficient cells. Based on these results, we hypothesize that inhibiting NMD-mediated degradation of the mutant GrnR493X mRNA would increase functional progranulin levels and thereby improve FTD-associated behavioral changes and neuropathology. We will test this hypothesis in the following two specific aims: Aim 1) Test in vivo efficacy of ASOs that block NMD-mediated degradation in the GrnR493X knock-in mouse model of FTD, and Aim 2) Design and test ASOs that target NMD-mediated degradation of the human GRNR493X mRNA. Completion of these studies will generate important preclinical data that provide insights into the utility of ASO-based NMD inhibition as a therapeutic approach for progranulin-deficient FTD.

项目概要 颗粒体蛋白前体是一种溶酶体分泌蛋白，含有多个富含半胱氨酸的颗粒体蛋白结构域；它是 精确的分子功能仍然未知。颗粒体蛋白前体 (GRN) 突变是额颞叶的病因 痴呆症 (FTD) 是一种毁灭性疾病，确诊后平均生存期为 3.8 年，且无法治愈 目前可用。由于颗粒体蛋白前体缺乏型 FTD 是一种单倍体不足的疾病，因此策略旨在 增加颗粒体蛋白前体水平是可行的治疗方法。我们最近生成了一个敲入小鼠 我们建立了含有常见患者无义突变 GRNR493X 的 FTD 模型，我们确定 无义介导的 mRNA 衰减 (NMD) 途径导致突变颗粒体蛋白前体显着减少 该小鼠模型以及含有 GRNR493X 突变的患者来源的成纤维细胞中的 mRNA 水平。 此外，我们的基于细胞的研究表明，颗粒体蛋白前体 R493X 突变蛋白在重新 在颗粒体蛋白前体缺陷的细胞中表达。基于这些结果，我们假设抑制 NMD 介导的 突变体 GrnR493X mRNA 的降解会增加功能性颗粒体蛋白前体水平，从而改善 FTD 相关的行为变化和神经病理学。我们将在下面两个例子中检验这个假设 具体目标： 目标 1) 测试 ASO 阻断 GrnR493X 中 NMD 介导的降解的体内功效 FTD 敲入小鼠模型，目标 2) 设计和测试针对 NMD 介导的 FTD 降解的 ASO 人 GRNR493X mRNA。这些研究的完成将产生重要的临床前数据，提供 深入了解基于 ASO 的 NMD 抑制作为颗粒体蛋白前体缺乏型 FTD 治疗方法的实用性。