Correction of Neurological Disease via Allele Specific Excision of Pathogenic Repeats

通过等位基因特异性切除致病重复序列来纠正神经系统疾病

基本信息

批准号：
10668665
负责人：
JENNIFER A DOUDNA
金额：
$ 468.35万
依托单位：
UNIVERSITY OF CALIFORNIA BERKELEY
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：
2023-05-15 至 2028-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10668665
关键词：
Alleles Amyotrophic Lateral Sclerosis Animal Model Animals Behavior Biodistribution Biological Assay Biotechnology C9ALS C9ORF72 CRISPR/Cas technology Cells Clinical Clustered Regularly Interspaced Short Palindromic Repeats Collaborations Convection Dedications Dependovirus Development Disease Disease model Dose Engineering Excision Feedback Fibroblasts Formulation Genes Genetic Guide RNA Haplotypes Heterozygote Histologic Human Huntington Disease In Vitro Induced pluripotent stem cell derived neurons Injections Investigational Therapies Lead Mendelian disorder Modeling Molecular Mus Mutation Nerve Degeneration Neurodegenerative Disorders Neurons Nucleotides Outcome Pathogenicity Patients Pharmacology Pharmacology and Toxicology Phenotype Preparation Process Production Property Rattus Reagent Research Research Personnel Research Project Grants Resources Ribonucleoproteins Rodent Model Route Serotyping Side Sum System Technology Testing Therapeutic Therapeutic Index Toxic effect Toxicology Transfection Triplet Multiple Birth Viral Virus Work animal tissue brain parenchyma brain tissue brain volume cell transformation design efficacy evaluation efficacy study experience genome editing improved in vivo innovation knock-down lead candidate manufacture mortality mouse model mutant nervous system disorder nonhuman primate novel peptide amphiphiles postmitotic pre-clinical prevent programs response screening synergism therapeutic genome editing tool

项目摘要

PROJECT SUMMARY / ABSTRACT A central promise of genome editing is its potential to treat monogenic disease. Despite early-stage clinical progress for CRISPR-Cas based approaches, monogenic neurodegenerative conditions and nucleotide triplet expansion disorders have not been a focus of any biotechnology company in this space. Our proposal brings together a team of academic investigators to develop a synergistic suite of first-in-class CRISPR-Cas based therapeutics for Huntington's disease (HD) and C9ORF72 amyotrophic lateral sclerosis (ALS). We will engineer and deploy an editing approach that excises, with IND-grade potency and mutant allele-selectivity, the disease- causing expansion repeat from human HTT and C9ORF72 loci, respectively. Our strategy is based on identifying alleles of commonly heterozygous SNPs that reside on the same haplotype as the disease-causing repeat expansion, and then engineering CRISPR-Cas9 for high selectivity of cleavage, on one or both sides of the mutant allele repeat, to drive its excision, with two tiers of delivery innovation. Our trailblazer project (Research Project 1, RP1) will develop an HD therapeutic by packaging mutant HTT-specific CRISPR-Cas9 into a newly developed serotype of adeno-associated virus (AAV) with robust and broad biodistribution in the brain parenchyma of nonhuman primates (NHP). We will implement an innovative strategy in which the CRISPR-Cas9 cassette temporally limits its own expression. We will identify and advance the preclinical lead composition through IND-enabling studies leveraging 3 dedicated Resource Cores to (i) assess molecular outcomes at the genetic level, (ii) administer reagents to animals and observe their behavior, and (iii) assess molecular and histological outcomes from cells and animal tissues. An Administrative Core led by experienced developers of genome editing-based therapeutics, will provide project-management support and lead on preparation of regulatory submissions, aiming to file an HD IND by program end. In RP2, we will apply the AAV-based excision approach to build a cognate experimental therapeutic for C9ORF72-driven ALS. Synergies with RP1 include CMC innovation to manufacture novel AAV, re-use of the self-regulating CRISPR-Cas cassette and virus harboring it, and regulatory feedback on IND-enabling pharmacology, toxicology, and biodistribution studies. We will advance RP2 through pre-IND. For RP3, we will establish a first-in-class, transformative paradigm for in vivo genome editing therapy by reformulating the preclinical lead CRISPR-Cas9 combination used in RP1 into a highly innovative “Cas9 RNP monoparticle” in which amphiphilic peptides deliver the gene editor to neurons upon injection. We will develop approaches for monoparticle manufacture to enable ex vivo and in vivo efficacy studies in HD models. Extensive synergies with RP1 project and comprehensive support by the RCs will enable us to advance this approach to pre-IND by program end. The sum total of this effort will establish a fundamentally new paradigm for in vivo genome editing applicable to all nucleotide repeat expansion disorders, and advance preclinical lead formulations for one disease, HD, to IND, and another such disease, C9ORF ALS, to pre-IND.

项目摘要 /摘要基因组编辑的核心承诺是它的治疗单基因疾病的潜力。尽管早期临床基于CRISPR-CAS的方法，单基因神经退行性条件和核苷酸三元组的进展扩张障碍并不是该领域任何生物技术公司的重点。我们的建议带来了组成一个学术研究人员团队，开发了一个基于一流的CRISPR-CAS的协同套件亨廷顿氏病（HD）和C9orf72肌萎缩性侧索硬化症（ALS）的治疗剂。我们将设计并采用一种编辑方法，该方法以印第安级和突变等位基因选择性进行消费，疾病 - 分别引起人类HTT和C9ORF72局部的扩展重复。我们的策略是基于确定的与引起疾病重复相同的单倍型的通常杂合SNP的等位基因扩展，然后在高度选择性的乳沟中进行工程crispr-cas9。突变等位基因重复出现，以引起其惊喜，并通过两级交付创新。我们的开拓者项目（研究项目1，RP1）将通过将突变体HTT特异性CRISPR-CAS9包装成新的HD疗法开发的腺相关病毒（AAV）的血清型在大脑中具有鲁棒和宽阔的生物分布非人类隐私（NHP）实质。我们将实施一种创新战略，其中CRISPR-CAS9 录音带暂时限制了自己的表达。我们将识别并推进临床前铅组成通过指定研究利用3个专用资源核心来评估分子结果遗传水平，（ii）对动物进行试剂并观察其行为，（iii）评估分子和来自细胞和动物组织的组织学结果。由经验丰富的开发人员领导的行政核心基于基因组编辑的治疗将提供项目管理支持，并在准备监管提交，旨在按计划结束提交高清IND。在RP2中，我们将采用基于AAV的惊喜为C9ORF72驱动的ALS建立同源实验疗法的方法。与RP1协同作用包括 CMC的创新以生产新颖的AAV，重新使用自我调节的CRISPR-CAS盒和病毒携带它，以及有关指定药理学，毒理学和生物分布研究的监管反馈。我们将通过预先启动RP2。对于rp3，我们将建立一个一等，体内的变革范式通过将RP1中用于临床前铅CRISPR-CAS9组合重新定义为基因组编辑疗法高度创新的“ CAS9 RNP单颗粒”，其中两亲性Petides将基因编辑器传递给神经元注射。我们将开发单颗粒制造的方法，以实现体内和体内效率高清模型的研究。与RP1项目的广泛协同作用，RCS的全面支持将启用我们要推进这种方法，以按计划结束。这项工作的总和将从根本上建立适用于所有核苷酸重复膨胀障碍的体内基因组编辑的新范式，并提高一种疾病的临床前铅配方，用于IND，另一种此类疾病C9orf ALS，以预先使用。