SCGE Disease Models Studies Supplement: Correction of RHO P23H adRP

SCGE 疾病模型研究补充：RHO P23H adRP 的校正

基本信息

批准号：
10619167
负责人：
RANDALL S PRATHER
金额：
$ 49.45万
依托单位：
UNIVERSITY OF MISSOURI-COLUMBIA
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-06-01 至 2024-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10619167
关键词：
Accounting Address Administrative Supplement Alleles Animals Blindness Canis familiaris Cells Clinic Clinical Clinical Trials Complement Cone Data Development Disease Disease Progression Disease model Dose Electroretinography Engineering Eye Family suidae Frequencies Future GRK1 gene Genes Genetic Transcription Genome Genomics Goals Gonadal structure Human Immunohistochemistry Injections Intervention Investigational Therapies Modality Modeling Morphology Mus Mutate Mutation Night Blindness Ophthalmic examination and evaluation Optical Coherence Tomography Organoids Patients Physiological Prevention Proteins Rare Diseases Reagent Research Personnel Retina Retinal Cone Retinitis Pigmentosa Rhodopsin Rod Safety Savings Site Specificity Structure Study models Technology Therapeutic Therapeutic Intervention Time TimeLine Tissues Transgenes Validation Variant Vertebrate Photoreceptors Viral Vision Work adeno-associated viral vector alternative treatment base clinically relevant effectiveness measure efficacy evaluation experimental study genome editing human model humanized mouse in vivo individualized medicine interest mutant next generation sequencing nonhuman primate nuclease participant enrollment pig genome porcine model preclinical evaluation preservation promoter response retinal rods somatic cell gene editing subretinal injection therapeutically effective tool treatment response vector

项目摘要

Project Summary The broad, long-term objective of this work is to establish a rubric to evaluate safety and efficacy of somatic cell genome editing technologies. These goals will be addressed in the context of an experimental therapy for retinitis pigmentosa caused by the RHO P23H variant. The specific aims of this proposal address refinement of the therapeutic window for this therapy, the minimum number of cells that must be edited to save vision, and the general safety of the therapeutic strategy. These aims will be met through the use of AAV5 to deliver an engineered meganuclease that specifically targets the dominant P23H allele while saving the wildtype allele that differs by only one base. This reagent will be evaluated in a validated, rapidly progressing swine model that harbors a human RHO P23H transgene. A previously optimized dose that produces a durable response will be delivered via subretinal injection and the animals will be followed for 32 weeks to repeatedly measure effectiveness over time via clinical eye exam, Full-Field Electroretinogram (ffERG), and Optical Coherence Tomography (OCT). At the end of the experiment, eyes will be examined via immunohistochemistry (IHC) to determine the regional, minimum number of rods that must be edited to save cone function and therefore to save vision. These experiments will be performed at two different stages in disease progression that are hypothesized to represent the last treatable state and the first non-treatable state so that guidance can be provided for criteria of enrollment of patients in a future clinical trial. In parallel, animals will be similarly treated and then sacrificed at two weeks post injection to determine the range of adjacent tissues to which the AAV5 vector will migrate. These animals will also be used to determine the level of editing that occurs in non-intended tissues, including the germline. All animals will be used to obtain data on general physiological response to the therapy.

项目概要这项工作的广泛、长期目标是建立一个评估安全性和安全性的标准体细胞基因组编辑技术的功效。这些目标将在由 RHO 引起的视网膜色素变性实验性治疗的背景 P23H 变体。该提案的具体目标涉及治疗的细化该疗法的窗口，必须编辑以挽救视力的最小细胞数量，以及治疗策略的总体安全性。这些目标将通过使用 AAV5 来提供专门针对显性 P23H 等位基因，同时保留仅相差一个碱基的野生型等位基因。这试剂将在经过验证的、快速进展的猪模型中进行评估，该模型包含人 RHO P23H 转基因。先前优化的剂量可产生持久的效果反应将通过视网膜下注射传递，并且动物将被跟踪 32 数周内通过临床眼科检查、全视野反复测量有效性视网膜电图 (ffERG) 和光学相干断层扫描 (OCT)。结束时实验中，将通过免疫组织化学 (IHC) 检查眼睛以确定区域，必须编辑的最小杆数以保存锥函数和因此要保存视力。这些实验将在两个不同阶段进行假设代表最后可治疗状态的疾病进展和第一个不可治疗的状态，以便可以为入组标准提供指导未来临床试验中的患者。与此同时，动物也会受到类似的对待，然后注射后两周处死以确定邻近组织的范围 AAV5 载体将迁移到其中。这些动物也将被用来确定在非预期组织（包括种系）中发生的编辑水平。所有动物将用于获得有关治疗的一般生理反应的数据。