Testing Optimal Gene Editor for an Alzheimer's CRISPR therapeutic.

测试阿尔茨海默病 CRISPR 疗法的最佳基因编辑器。

• 批准号: 10746716
• 负责人: Subhojit Roy
• 金额: $ 211.28万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 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 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10746716
• 关键词: Advisory Committees; Alzheimer' s Disease; Alzheimer' s disease therapeutic; Alzheimer' s disease therapy; Amyloid; Amyloid beta-Protein; Amyloid beta-Protein Precursor; Attenuated; Biological Assay; Brain; CRISPR therapeutics; CRISPR/Cas technology; Cell surface; Cells; Clinic; Clinical; Clinical Paths; Clinical Trials; Clustered Regularly Interspaced Short Palindromic Repeats; Consensus; Consultations; Deposition; Disease; Dose; Down Syndrome; Early Onset Alzheimer Disease; Endosomes; Equilibrium; Etiology; Event; Exons; Eye diseases; FDA approved; Failure; Future; Gene Proteins; Genes; Genetic; Genome; Goals; Guide RNA; Hand; Human; Human Amyloid Precursor Protein; Human Genetics; Injections; Kinetics; Knock-in; Knock-in Mouse; Lead; Letters; Measures; Mediating; Mission; Modality; Mouse Protein; Mus; Nerve Regeneration; Neurofibrillary Tangles; Neurons; Organoids; Outcome; Pathologic; Pathway interactions; Persons; Pharmaceutical Preparations; Phase I/II Trial; Physiological; Physiology; Positioning Attribute; Protein C; Protein Fragment; Protein Truncation; Proteins; Proteolysis; Public Health; Quantitative Reverse Transcriptase PCR; RNA; Recommendation; Research; Research Personnel; Retina; Role; Safety; Surface; Testing; Therapeutic; Time; Transcript; Transmembrane Domain; United States National Institutes of Health; Variant; adeno-associated viral vector; base editing; beta-site APP cleaving enzyme 1; cost; efficacy testing; experimental study; extracellular; genome-wide; in vivo; induced pluripotent stem cell; meetings; mouse model; neuroprotection; nonhuman primate; novel; novel therapeutics; prevent; response; secretase; tau Proteins; tau aggregation; therapeutic genome editing; translation to humans

Summary/Abstract: The overall goal of this U01 proposal is to advance a unique CRISPR-based therapeutic for Alzheimer’s disease (AD) towards the pre-IND stage, by determining the best genome-editor/gRNA combination that can be ultimately used in clinical trials. Though there is an enormous unmet need for developing therapeutics for AD, we do not have drugs that can unequivocally slow down the relentless course of AD. We have developed a gene-editing based strategy targeting APP – a gene with a central and indisputable role in AD; strongly supported by human genetics. We do not eliminate the APP gene, but edit out a pentapeptide YENPTY endocytic domain at the C- terminus of APP, which blocks the entire pathologic APP β-cleavage pathway (including β-amyloid). The transmembrane domain and the N-terminus remains intact, and the ∆C APP is retained on the cell surface, leading to an increase in APP α-cleavage, which in turn upregulates neuroprotective and neuroregenerative APP fragments. Thus, our strategy shifts the balance of APP cleavage from pathologic to physiologic, without eliminating the gene. Since the targeted events are upstream, our strategy should be applicable to forms of AD – sporadic and familial – and also to early-onset AD in Down syndrome, where APP triplication on Ch.21 invariably causes AD. So far, we have used the classical SaCas9 to demonstrate safety and efficacy of our approach in vivo, but SaCas9 is too big to fit into a single AAV, which will be necessary for ultimate delivery in humans. Moreover, since there is no FDA-approved gene-editing clinical trial in the brain yet, and the best genome-editor is unknown. Here, we propose to first test two promising small genome-editors (and corresponding gRNAs) that can fit into a single AAV – SaCas9 and NmCas9 – using genome-scale on- and off- target assays (Aim 1). The best Lead-Editor/gRNA combinations emerging from these experiments will be tested for efficacy, safety, predictability, and durability in human brain organoids and a novel APP mouse model where the entire mouse APP was replaced with human APP (Aim 2). Collectively, these studies will not only clarify the best genome-editor/RNA combination for editing the APP gene, but also help future trials in the brain gene- editing space by determining the best editor for brain-relevant applications. Towards the end of our projects, we propose to hold an INTERACT meeting with the FDA – in consultation with our Clinical/Translational Advisory Team – to get regulatory input and consensus on pre-IND NHP studies. Upon completion, our studies will not only offer a novel therapeutic in an arena of enormous unmet need and litany of failures, but also provide a clinical path for future gene-editing efforts in this therapeutic space.

摘要/摘要： 该U01提案的总体目标是为阿尔茨海默氏病进行基于CRISPR的独特疗法 （AD）通过确定最终可以是最终的基因组/GRNA组合，是通过确定最终的基因组和GRNA组合的（AD） 用于临床试验。尽管对开发AD的治疗巨大不满意，但我们没有 有可以明确地减慢无端AD过程的药物。我们已经开发了一个基因编辑 针对应用程序的基于策略 - 在AD中具有中心和无可争议的作用的基因；由人类强烈支持 遗传学。我们不会消除APP基因，而是在c-上编辑五肽的五肽内吞域 APP的末端，它阻止了整个病理APPβ-裂解途径（包括β-淀粉样蛋白）。这 跨膜结构域和N末端保持完整，并且∆C应用保留在细胞表面， 导致APPα-裂解的增加，这反过 碎片。这，我们的策略将应用程序裂解的平衡从病理学转变为生理学，而没有 消除基因。由于目标事件是上游，因此我们的策略应适用于广告形式 - 零星和家庭 - 以及唐氏综合症的早期发作广告 总是引起广告。到目前为止，我们已经使用经典SACAS9来证明我们的安全性和效率 在体内接近，但是SACAS9太大了，无法适应一个AAV，这对于最终交付是必要的 人类。此外，由于尚无大脑中FDA批准的基因编辑临床试验，也是最好的 基因组编辑未知。在这里，我们建议首先测试两个有希望的小型基因组编辑器（和 相应的grnas）可以使用基因组尺度的on和off-适合单个AAV - SACAS9和NMCAS9 目标测定（目标1）。将测试这些实验的最佳主编码器/GRNA组合 为了效率，安全性，可预测性和耐用性，人类脑器官和一种新型的App Mouse模型，其中 整个鼠标应用程序被人类应用替换（AIM 2）。总的来说，这些研究不仅会阐明 最好的基因组编辑器/RNA组合用于编辑APP基因，但也有助于将来的大脑基因试验 通过确定与大脑相关应用的最佳编辑器来编辑空间。在我们的项目结束时，我们 提议与FDA举行互动会议 - 与我们的临床/翻译咨询协商 团队 - 在预先印度NHP研究上获得监管投入和共识。完成后，我们的学习不会 仅在巨大的未满足需求和失败的竞技场中提供一种新颖的疗法，但也提供 在此治疗空间中，未来基因编辑工作的临床路径。