CRISPR gene therapies targeting tau in Alzheimer's disease and tauopathies

针对阿尔茨海默病和 tau 病中 tau 蛋白的 CRISPR 基因疗法

• 批准号: 10752745
• 负责人: Todd Jonathan Cohen
• 金额: $ 42.41万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10752745
• 关键词: Alzheimer' s Disease; Alzheimer' s disease brain; Alzheimer' s disease model; Alzheimer' s disease patient; Alzheimer' s disease related dementia; Alzheimer' s disease therapeutic; Antisense Oligonucleotides; Biochemical; Brain; CRISPR/Cas technology; Cell Line; Clustered Regularly Interspaced Short Palindromic Repeats; Cognitive; Complex; DNA; DNA Damage; Data; Defect; Dementia; Development; Disease; Disease Progression; Effectiveness; Epitopes; Flowers; Functional disorder; Genome; Genomic DNA; Goals; Guide RNA; Human; Human Cell Line; Immunotherapy; Impaired cognition; In Vitro; Injections; Lentivirus; Libraries; MAPT gene; Measures; Mediating; Memory Loss; Methods; Modality; Modeling; Mus; Nerve Degeneration; Nervous System; Neurons; Onset of illness; Pathology; Patients; Penetrance; Penetration; Peripheral; Pick Disease of the Brain; Progressive Supranuclear Palsy; Property; Protein Isoforms; RNA; Serotyping; Site; Specificity; Synapses; System; Tauopathies; Technology; Testing; Therapeutic; Time; Transcript; Validation; Variant; behavioral study; chemical stability; corticobasal syndrome; efficacy testing; gene therapy; histological studies; improved; in vivo; innovation; interest; intravenous injection; knock-down; mouse model; multi-electrode arrays; novel; preclinical efficacy; targeted treatment; tau Proteins; tau aggregation; therapeutic evaluation; tool; tumorigenesis

PROJECT SUMMARY / ABSTRACT The MAPT gene encoding the tau protein is an ideal target for Alzheimer’s disease (AD) therapeutics since it forms hallmark pathology in all AD patients. In mice, tau aggregation drives disease progression, while depleting tau leads to cognitive improvements, suggesting targeting tau may benefit human patients as well. A major question has been how to achieve this goal. CRISPR technology (Clustered Regularly Interspaced Short Palindromic Repeats) has blossomed in recent years, and we can now leverage this approach to specifically deplete tau in neurons. By targeting different tau isoforms with site-specific guide RNAs of interest, one can conceivably target the specific tau variants that are present in 3R-tauopathies (e.g., Pick’s disease), 4R tauopathies (e.g., corticobasal syndrome), or mixed 3R/4R tauopathies (e.g., AD patients). Thus, we turned to a newly developed CRISPR/Cas13 technology, which allows targeting and depletion of RNA transcripts rather than DNA itself, the latter of which can have unwanted effects on the genome including DNA damage and even tumorigenesis. CRISPR/Cas13 technology could provide new opportunities for therapeutic tau reduction and potentially overcome many of the limitations that have plagued tau depletion strategies. In fact, our preliminary findings show that we can computationally predict which guides will successfully target and deplete tau, and we demonstrated this in a human cell line and primary human neurons, suggesting we have found an optimal pipeline for guide prediction, development, and validation. Therefore, we have an exciting opportunity to develop an effective, non-invasive tool for tau gene therapy in the nervous system (CNS). In Aim-1, we will develop and refine the CRISPR/Cas13 system to target tau and will comprehensively identify the most optimal guide RNAs that target either 3R-tau, 4R-tau, or total tau. We will deliver those guides to neurons and determine their effectiveness and ability to restore normal neuronal function. In Aim-2, we will perform preclinical efficacy testing and determine whether our most optimal CRISPR/Cas13 complexes have therapeutic potential in AD model mice. By delivering tau-targeting guides peripherally using brain-penetrant AAV methods, we will evaluate their therapeutic impact by performing a battery of biochemical, histological, and behavioral studies at either early (preventative) or late (therapeutic) stages of disease progression. This study is innovative and significant since it will both develop the CRISPR technology needed to target tau, and also advance gene therapy approaches to deplete tau in humans including AD and other dementia patients. The clear implications are that targeting and reducing tau levels, even modestly, could provide a new treatment modality for dementia patients.

项目概要/摘要 编码 tau 蛋白的 MAPT 基因是阿尔茨海默病 (AD) 治疗的理想靶点，因为它 形成所有 AD 患者的标志性病理学。在小鼠中，tau 蛋白聚集会驱动疾病进展，而 消耗 tau 蛋白可以改善认知能力，这表明针对 tau 蛋白的靶向治疗也可能使人类患者受益。一个 主要问题是如何实现这一目标。 CRISPR技术（成簇规则间隔短 Palindromic Repeats）近年来蓬勃发展，我们现在可以利用这种方法来专门 耗尽神经元中的 tau 蛋白。通过使用感兴趣的位点特异性引导 RNA 靶向不同的 tau 亚型，人们可以 可以想象，针对 3R-tau 病（例如匹克病）、4R 中存在的特定 tau 变体 tau蛋白病（例如，皮质基底节综合征），或混合 3R/4R tau蛋白病（例如，AD 患者）。因此，我们转向 新开发的 CRISPR/Cas13 技术，可以靶向并消除 RNA 转录本 DNA 本身可能会对基因组产生不良影响，包括 DNA 损伤，甚至 肿瘤发生。 CRISPR/Cas13技术可以为治疗性tau蛋白减少和治疗提供新的机会 有可能克服困扰 tau 消耗策略的许多限制。事实上，我们初步 研究结果表明，我们可以通过计算预测哪些导向器将成功靶向并耗尽 tau 蛋白，并且 我们在人类细胞系和原代人类神经元中证明了这一点，这表明我们已经找到了最佳的 用于指南预测、开发和验证的管道。因此，我们有一个激动人心的机会 开发一种有效的、非侵入性的神经系统 (CNS) tau 基因治疗工具。在目标 1 中，我们将 开发和完善针对 tau 的 CRISPR/Cas13 系统，并将全面识别最优化的 靶向 3R-tau、4R-tau 或总 tau 的引导 RNA。我们将把这些指南传递给神经元 确定它们恢复正常神经元功能的有效性和能力。在Aim-2中，我们将执行 临床前功效测试并确定我们最优化的 CRISPR/Cas13 复合物是否具有 AD模型小鼠的治疗潜力。通过使用脑渗透剂在外周提供 tau 靶向导向 AAV 方法，我们将通过进行一系列生化、组织学、 以及疾病进展早期（预防性）或晚期（治疗性）阶段的行为研究。这 这项研究具有创新性和意义，因为它将开发靶向 tau 所需的 CRISPR 技术，并且 还推进了基因治疗方法，以消除人类（包括 AD 和其他痴呆症患者）的 tau 蛋白。 明显的含义是，瞄准并降低 tau 水平，即使是适度的，也可以提供一种新的治疗方法 痴呆症患者的治疗方法。