Precision Base Editing for the Treatment of Motor Neuron Diseases

精确碱基编辑治疗运动神经元疾病

基本信息

批准号：
10703727
负责人：
Mandana Arbab
金额：
$ 24.9万
依托单位：
BOSTON CHILDREN'S HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-08-01 至 2026-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10703727
关键词：
Acceleration Address Adult Affect Alleles Amyotrophic Lateral Sclerosis Animal Model Antisense Oligonucleotides Award BCAR1 gene Cell Death Central Nervous System 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 Late-Onset Disorder Learning Libraries Mammalian Cell Mentorship Methods Minor Modeling Molecular Abnormality Motor Neuron Disease Motor Neurons Mus Mutation Neonatal 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 Testing Therapeutic Therapeutic Intervention Tissues Translating Treatment Efficacy United States National Institutes of Health Validation Variant Work Writing base editing base editor 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 machine learning prediction meetings member molecular pathology molecular phenotype mouse model mutant novel predictive modeling progressive neurodegeneration screening skills success symposium therapeutic development therapeutic genome editing tool transversion mutation

项目摘要

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），例如脊柱肌肉萎缩（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次要遗传异常引起。由于这些疾病中的每一种都是罕见的，并且分子病理如此潜水和未解决，因此很难设想针对每种单独量身定制的治疗干预措施，或一种单独的治疗方法来有效治疗它们。当前的基因组编辑工具很容易针对靶向离散的基因组局部，并且受影响的MND的组织在很大程度上相似。这是针对每个突变的MND的一般基因组编辑理论策略，可以满足这一迫切需求。基础编辑工具从理论上可以纠正任何C：g> t：A和A：T> G：C转换，但是确定基础编辑效率和精度的因素尚不完全了解，并且在给定基因座的编辑结果通常是不可预测的。为了开发大量基因座的基础编辑校正策略，通过多种基础编辑器变体进行筛选 - 当前的任何二次脱氨酶酶和> 15个CAS蛋白的排列以及计数 - 以及每个目标的SGRNA组合都被禁止。需要清楚地了解CAS蛋白，脱氨酶和序列的编辑结果决定剂，以促进基础编辑策略的设计。我们打算开发一般的工作流程，以设计有效的因果MND SNP的基础编辑策略，并将这些工具提供给MND受影响的组织。 SMA是一个单基因的MND，具有明确的遗传原因，并且具有人类因果基因的动物模型忠实地概括了SMA患者的疾病表型。成功开发有效且安全的SMA基因组编辑治疗将有助于开发针对其他遗传MND的类似基因组编辑疗法，包括某些形式的家庭ALS。在这个项目中，我们将（1）创建一个基础编辑的计算预测模型，以促进有效的基础编辑策略的设计；（2）制定方案，以有效地为小鼠相关时机提供基础编辑疗法以实现基因组编辑；（3）使用该管道优化基础编辑疗法来挽救小鼠中的SMA，并为其他因果MND突变开发基因组编辑疗法，首先关注北美最常见的单点突变因果关系，SOD1A4V。我将利用大型研究所的世界一流基因组学环境，以及斯坦利精神病学研究中心的AAV交付专业知识，以实现这些目标和发展技能，这将帮助我继续作为独立研究者的这项工作。通过关于赠款和数据可视化的心态会议和课程，我将提高我的科学通讯技能，以便我可以成功竞争作为教职员工的额外NIH R01和R21资金。通过参加会议并发表我的工作，我打算将自己确立为MND治疗基因组编辑领域的领导者。