Precision base editing for the treatment of motor neuron diseases

精准碱基编辑治疗运动神经元疾病

• 批准号: 10301562
• 负责人: Mandana Arbab
• 金额: $ 12.65万
• 依托单位: BROAD INSTITUTE, INC.
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-01 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10301562
• 关键词: Address; Adult; Affect; Alleles; Amyotrophic Lateral Sclerosis; Animal Model; Antisense Oligonucleotides; Award; BCAR1 gene; Cell Death; Chimeric Proteins; Clinic; Communication; Complex; DNA Sequence Alteration; Data; Deaminase; Development; Disease; Disease model; Disease-Free Survival; Environment; Enzymes; Faculty; Foundations; Funding; Future; Genes; Genetic; Genome; Genomics; Genotype; Goals; Grant; Human; In Vitro; Individual; Injections; Institutes; Late-Onset Disorder; Learning; Libraries; Machine Learning; Mammalian Cell; Mentorship; Methods; Minor; Modeling; Molecular Abnormality; Motor Neuron Disease; Motor Neurons; Mus; Mutation; Neonatal; Neuraxis; Neurodegenerative Disorders; North America; Nucleotides; Other Genetics; Outcome; Pathogenicity; Patients; Peripheral; Phenotype; Point Mutation; Positioning Attribute; Procedures; Protocols documentation; Publishing; Research; Research Personnel; Route; Safety; Science; Serotyping; Spinal Muscular Atrophy; System; Techniques; Therapeutic; Therapeutic Intervention; Tissues; Translating; Treatment Efficacy; United States National Institutes of Health; Validation; Variant; Work; Writing; base; causal variant; community based research; data visualization; design; disease phenotype; disease-causing mutation; embryonic stem cell; experimental study; familial amyotrophic lateral sclerosis; genetic variant; genome editing; genomic locus; improved; in vivo; meetings; member; molecular pathology; molecular phenotype; mouse model; mutant; novel; predictive modeling; predictive test; programs; progressive neurodegeneration; screening; skills; success; symposium; therapeutic development; therapeutic genome editing; tool; transversion mutation

Motor neuron diseases (MNDs) such as spinal muscular atrophy (SMA) and amyotrophic lateral sclerosis (ALS) are a class of progressive neurodegeneration disorders, often caused by minor genetic abnormalities to which motor neurons are particularly vulnerable. Because each of these diseases is rare and the molecular pathologies so diverse and unresolved, it is hard to envision a therapeutic intervention tailored to each individually, or a single treatment to effectively treat them all. Current genome editing tools are easily programmable to target discrete genomic loci and affected tissues of MNDs are largely similar. Thus, a general genome editing therapeutic strategy for MNDs fitted to each mutation could meet this urgent need. Base editing tools can theoretically correct any C:G>T:A and A:T>G:C transition, yet the factors that determine efficiency and precision of base editing are not fully understood, and editing outcomes at a given locus are frequently unpredictable. For the development of base editing correction strategies at a great number of loci, screening through a multitude of base editor variants – currently any permutation of >10 deaminase enzymes and >15 Cas proteins, and counting – and sgRNA combinations for every target is prohibitive. A clear understanding of Cas protein, deaminase, and sequence determinants of editing outcomes is needed to facilitate the design of base editing strategies. We intend to develop a general workflow to design effective base editing strategies for causal MND SNPs and deliver these tools to MND affected tissues. SMA is a monogenic MND with a well-defined genetic cause, and animal models harboring the human causal gene that faithfully recapitulate disease phenotypes of SMA patients. Successful development of an effective and safe SMA genome editing treatment will assist the development of similar genome editing therapeutics for other genetic MNDs, including some forms of familial ALS. In this project we will (1) create a computational predictive model of base editing to facilitate the design of effective base editing strategies; (2) develop protocols to efficiently deliver base editing therapeutics to disease relevant tissues in mice to enable genome editing; and (3) use this pipeline to optimize a base editing therapeutic to rescue SMA in mice, and develop genome editing therapeutics for other causal MND mutations, focusing first on the most common single point mutation causal to ALS in North America, SOD1A4V. I will take advantage of the world-class genomics environment of the Broad Institute and the expertise in AAV-delivery at the Stanley Center for Psychiatric Research to realize these goals and develop skills that will help me to continue this work as an independent investigator. Through mentorship meetings and courses on grant-writing and data visualization I will improve my science-communication skills so that I may compete successfully for additional NIH R01 and R21 funding as a faculty member. By attending conferences and publishing my work I intend to establish myself as leader in the field of therapeutic genome editing for MND.

运动神经元疾病（MND）如脊髓性肌萎缩症（SMA）和肌萎缩性侧索硬化症（ALS）是一类进行性神经变性疾病，通常由运动神经元特别容易受到的轻微遗传异常引起。由于这些疾病中的每一种都是罕见的，分子病理学如此多样且尚未解决，因此很难想象针对每一种疾病单独进行治疗干预，或者单一治疗方法可以有效地治疗所有疾病。目前的基因组编辑工具易于编程以靶向离散的基因组基因座，并且MND的受影响组织在很大程度上相似。因此，针对适合每个突变的MND的通用基因组编辑治疗策略可以满足这一迫切需求。 碱基编辑工具理论上可以纠正任何C：G>T：A和A：T>G：C转换，但决定碱基编辑效率和精度的因素尚未完全理解，并且给定位点的编辑结果通常是不可预测的。为了在大量基因座处开发碱基编辑校正策略，通过多个碱基编辑器变体（目前>10种脱氨酶和>15种Cas蛋白的任何排列，以及计数）和针对每个靶标的sgRNA组合进行筛选是禁止的。需要清楚地了解Cas蛋白、脱氨酶和编辑结果的序列决定因素，以促进碱基编辑策略的设计。我们打算开发一个通用的工作流程来设计有效的碱基编辑策略的因果MND单核苷酸多态性，并提供这些工具的MND受影响的组织。 SMA是一种单基因MND，具有明确的遗传原因，动物模型携带人类致病基因，忠实地再现了SMA患者的疾病表型。成功开发有效和安全的SMA基因组编辑治疗将有助于开发用于其他遗传MND的类似基因组编辑疗法，包括某些形式的家族性ALS。 在这个项目中，我们将（1）创建一个碱基编辑的计算预测模型，以促进有效的碱基编辑策略的设计;（2）开发协议，以有效地将碱基编辑疗法递送到小鼠的疾病相关组织，以实现基因组编辑;以及（3）使用该管道来优化碱基编辑治疗剂以拯救小鼠中的SMA，并为其他致病性MND突变开发基因组编辑疗法，首先关注北美ALS最常见的单点突变SOD 1A 4V。 我将利用布罗德研究所世界一流的基因组学环境和斯坦利精神病学研究中心在AAV递送方面的专业知识来实现这些目标，并发展技能，以帮助我作为独立研究者继续这项工作。通过指导会议和课程补助金写作和数据可视化，我将提高我的科学沟通技巧，使我可以成功地竞争额外的NIH R 01和R21资金作为一名教师。通过参加会议和发表我的工作，我打算将自己确立为MND治疗性基因组编辑领域的领导者。