Optimization of an in vivo base editing strategy to treat SOD1-linked ALS

优化治疗 SOD1 相关 ALS 的体内碱基编辑策略

• 批准号: 10351588
• 负责人: Thomas Gaj
• 金额: $ 86.99万
• 依托单位: UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-12-23 至 2026-11-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10351588
• 关键词: Address; Adverse effects; Amyotrophic Lateral Sclerosis; Antisense Oligonucleotides; Brain; CRISPR/Cas technology; Cells; Clustered Regularly Interspaced Short Palindromic Repeats; DNA; Dependovirus; Disease; Disease Progression; Dose; Drug Kinetics; Ensure; Financial Hardship; Foundations; Future; Gene Expression; Gene Silencing; Genes; Goals; Half-Life; Histologic; Immune response; Immunologics; Immunology; Infusion procedures; Interdisciplinary Study; Lead; Link; Macaca fascicularis; Messenger RNA; Methods; Modality; Modeling; Molecular; Motor Neurons; Mutation; Nervous system structure; Nonhomologous DNA End Joining; Outcome; Paralysed; Pathway interactions; Patients; Probability; Production; Program Development; Property; Proteins; RNA Processing; Regimen; Reporting; Research; Risk; Running; Safety; Severities; Single base substitution; Specificity; Spinal Cord; Technology; Therapeutic Effect; Time; Toxicology; Variant; amyotrophic lateral sclerosis therapy; base; base editing; base editor; effective therapy; gain of function mutation; gene therapy; illness length; in vivo; innovation; meetings; mouse model; mutant; nervous system disorder; neurotoxic; nonhuman primate; novel; novel strategies; nuclease; preclinical development; programs; recruit; scaffold; superoxide dismutase 1; therapeutic candidate; therapy development; virtual

PROJECT SUMMARY Amyotrophic lateral sclerosis (ALS) is a rapidly progressive, paralytic and ultimately fatal disease characterized by the selective loss of motor neurons in the spinal cord and brain. The overarching objective of this application to the Optimization Track of the CREATE Bio Program is to refine the safety and efficacy of a gene therapy that we developed for forms of ALS caused by toxic, gain-of-function mutations in superoxide dismutase 1 (SOD1), which account for up to 20% of all familial cases of the disease. Specifically, we have developed an approach to inactivate the production of the mutant SOD1 protein in vivo using CRISPR base editing, a gene- editing modality capable of introducing precise base substitutions in DNA, but without the requirement for a mutagenic DNA break, thereby overcoming a major safety hurdle facing the implementation of traditional gene- editing nucleases. In particular, when delivered to the spinal cord via adeno-associated virus, our SOD1- targeting base-editing platform prolonged survival and markedly slowed the progression of disease in a highly aggressive mouse model of SOD1-linked ALS. Importantly, as opposed to current strategies for silencing SOD1, which target SOD1 mRNA and can have a transient effect that requires a lifetime of redosing or can risk saturating endogenous RNA processing pathways, which could then lead to adverse effects, our approach harnesses a highly precise DNA editing pathway to permanently turn-off the production of mutant SOD1 and involves only a single treatment. Thus, because of its strengths and in vivo efficacy, we now aim to refine this strategy for the ultimate goal of developing a gene therapy for ALS. Specifically, by optimizing its targeting specificity, its editing capabilities, its pharmacokinetics and its safety, which we will ensure via the introduction of a self-inactivating functionality that facilitates its clearance from cells, we will develop a highly optimized SOD1-targeting CRISPR base editing platform that can be used to permanently and effectively treat SOD1-linked ALS. Thus, by capitalizing on: (1) a highly innovative DNA editing technology that has the capabilities to overcome the limitations of gene-silencing and (2) a multidisciplinary research team with complementary expertise in ALS, AAV delivery, gene-editing and immunology, this project will result in not only an optimized therapeutic candidate for ALS, a devastating, debilitating and currently incurable disorder with few effective treatment options, but also lay the foundation for using base editing to safely treat neurological disorders.

项目摘要 肌萎缩侧索硬化症（ALS）是一种快速进行性、麻痹性和最终致命的疾病，其特征在于： 脊髓和大脑中运动神经元的选择性丧失。本申请的首要目标是 CREATE Bio计划的优化轨道是改进基因治疗的安全性和有效性 我们开发了针对由超氧化物歧化酶1中的毒性、功能获得性突变引起的ALS形式的药物， （SOD 1），占所有家族性病例的20%。具体来说，我们开发了一种 使用CRISPR碱基编辑，一种基因- 能够在DNA中引入精确的碱基取代，但不需要 诱变DNA断裂，从而克服了传统基因工程实施所面临的一个主要安全障碍， 编辑核酸酶。特别是，当通过腺相关病毒传递到脊髓时，我们的SOD 1- 靶向碱基编辑平台延长了生存期，并显著减缓了疾病的进展， SOD 1连锁ALS的侵袭性小鼠模型。 重要的是，与目前沉默SOD 1的策略相反，目前沉默SOD 1的策略靶向SOD 1 mRNA，并且可以具有抑制SOD 1表达的作用。 需要终生再给药或可能有使内源性RNA加工饱和的风险的短暂效应 途径，这可能会导致不利影响，我们的方法利用了高度精确的DNA编辑， 这是一种永久关闭突变SOD 1产生的途径，仅涉及单次治疗。因此，在本发明中， 由于其优势和体内疗效，我们现在的目标是完善这一战略的最终目标， 开发ALS的基因疗法具体地说，通过优化其靶向特异性、编辑能力、 药代动力学及其安全性，我们将通过引入自失活功能来确保， 促进其从细胞中清除，我们将开发一种高度优化的SOD 1靶向CRISPR碱基编辑 该平台可用于永久有效地治疗与SOD 1相关的ALS。因此，通过利用：（1）a 高度创新的DNA编辑技术，能够克服基因沉默的局限性 和（2）一个多学科研究团队，在ALS、AAV递送、基因编辑和 免疫学，该项目将导致不仅是一个优化的治疗候选人ALS，一个毁灭性的， 使人衰弱和目前无法治愈的疾病，几乎没有有效的治疗选择，但也奠定了基础， 使用碱基编辑来安全地治疗神经系统疾病。