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

基本信息

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
项目类别：
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&apos s Disease Alzheimer&apos s disease brain Alzheimer&apos s disease model Alzheimer&apos s disease patient Alzheimer&apos s disease related dementia Alzheimer&apos 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

项目摘要

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.

项目摘要 /摘要编码tau蛋白的MAPT基因是阿尔茨海默氏病（AD）疗法的理想靶标在所有AD患者中形成标志性病理。在小鼠中，tau聚集驱动疾病进展，而耗尽的tau导致认知改善，这表明靶向tau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（聚类定期间隔短 palindromic重复序列）近年来已经开花了，现在我们可以利用这种方法专门在神经元中耗尽tau。通过使用感兴趣的特定地点指南RNA靶向不同的TAU同工型，一个人可以可以想象，可以针对3R-Tauopath（例如，Pick的疾病）中存在的特定TAU变体，4R tauopathies（例如，皮质综合征）或混合3R/4R tauopathies（例如AD患者）。那我们转向了新开发的CRISPR/CAS13技术，允许靶向和部署RNA成绩单比DNA本身，后者对基因组有不必要的影响，包括DNA损伤甚至肿瘤发生。 CRISPR/CAS13技术可以为减少治疗性TAU提供新的机会有可能克服困扰Tau耗竭策略的许多局限性。实际上，我们的初步调查结果表明，我们可以在计算上预测哪些指南将成功靶向和复制tau，并且我们在人类细胞系和原发性人神经元中证明了这一点，这表明我们发现了一个最佳指南预测，开发和验证的管道。因此，我们有一个令人兴奋的机会为神经系统（CNS）中的tau基因治疗开发有效的无创工具。在AIM-1中，我们将开发和完善CRISPR/CAS13系统以瞄准tau，并将全面确定最佳的指导靶向3R-TAU，4R-TAU或TAU的RNA。我们将向神经元传递这些指南，确定它们的有效性和恢复正常神经元功能的能力。在AIM-2中，我们将执行临床前效率测试并确定我们最佳的CRISPR/CAS13复合物是否具有 AD模型小鼠的治疗潜力。通过使用脑渗透剂外围传递tau靶向指南 AAV方法，我们将通过执行一系列生化，组织学，以及疾病进展的早期（预防性）或晚期（治疗）阶段的行为研究。这研究具有创新性和意义还可以推进基因治疗方法在人类中转tau，包括AD和其他痴呆症患者。明显的含义是，靶向和降低tau水平，甚至适度，都可以提供新的治疗方法痴呆症患者的方式。