Enhancing cone survival in retinitis pigmentosa through cell-specific therapeutic CRISPR editing of a roxadustat target

通过 roxadustat 靶标的细胞特异性治疗性 CRISPR 编辑来增强色素性视网膜炎中视锥细胞的存活

• 批准号: 10421156
• 负责人: JAMES Bryant HURLEY
• 金额: $ 54.55万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-01 至 2027-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10421156
• 关键词: Ablation; Adult; Aerobic; Affect; Age; Anemia; Blindness; Blood capillaries; CRISPR therapeutics; Carbon; Cell Shape; Cells; Cessation of life; Choroid; Citric Acid Cycle; Clinical; Clinical Trials; Clustered Regularly Interspaced Short Palindromic Repeats; Cone; Coupling; Data; Dependovirus; Disease; Doctor of Philosophy; Drug Kinetics; Enzymes; European; Functional disorder; Funding; Future; Gene Mutation; Genes; Genetic; Glucose; Glycolysis; Goals; Human; Inherited; Interphase Cell; Investigational Drugs; Investigational New Drug Application; Knock-out; Knowledge; Lead; Mediating; Medicine; Metabolic; Metabolic Pathway; Metabolism; Mission; Modeling; Modification; Mus; Mutation; Neurodegenerative Disorders; Oral; Organoids; Outcome; Pathogenesis; Patients; Pharmacology; Phase; Photoreceptors; Process; Procollagen-Proline Dioxygenase; Production; Proteins; Public Health; RPE65 protein; Research; Retina; Retinal Cone; Retinal Diseases; Retinal Dystrophy; Retinitis Pigmentosa; Rod; Role; Safety; Source; Starvation; Structure of retinal pigment epithelium; Subgroup; Teenagers; Testing; Therapeutic; Tissues; Toxic effect; Translating; Treatment Cost; Ubiquitination; Vertebrate Photoreceptors; Vision; Vision research; aerobic glycolysis; antagonist; base; clinically significant; cost; experimental study; gene therapy; hemangioblastoma; innovation; metabolome; mouse model; normoxia; novel; novel therapeutics; photoreceptor degeneration; preservation; promoter; repaired; response; retinal rods; therapeutic genome editing; therapeutic target; therapeutically effective; tool; vector; Ablation; Adult; Aerobic; Affect; Age; Anemia; Blindness; Blood capillaries; CRISPR therapeutics; Carbon; Cell Shape; Cells; Cessation of life; Choroid; Citric Acid Cycle; Clinical; Clinical Trials; Clustered Regularly Interspaced Short Palindromic Repeats; Cone; Coupling; Data; Dependovirus; Disease; Doctor of Philosophy; Drug Kinetics; Enzymes; European; Functional disorder; Funding; Future; Gene Mutation; Genes; Genetic; Glucose; Glycolysis; Goals; Human; Inherited; Interphase Cell; Investigational Drugs; Investigational New Drug Application; Knock-out; Knowledge; Lead; Mediating; Medicine; Metabolic; Metabolic Pathway; Metabolism; Mission; Modeling; Modification; Mus; Mutation; Neurodegenerative Disorders; Oral; Organoids; Outcome; Pathogenesis; Patients; Pharmacology; Phase; Photoreceptors; Process; Procollagen-Proline Dioxygenase; Production; Proteins; Public Health; RPE65 protein; Research; Retina; Retinal Cone; Retinal Diseases; Retinal Dystrophy; Retinitis Pigmentosa; Rod; Role; Safety; Source; Starvation; Structure of retinal pigment epithelium; Subgroup; Teenagers; Testing; Therapeutic; Tissues; Toxic effect; Translating; Treatment Cost; Ubiquitination; Vertebrate Photoreceptors; Vision; Vision research; aerobic glycolysis; antagonist; base; clinically significant; cost; experimental study; gene therapy; hemangioblastoma; innovation; metabolome; mouse model; normoxia; novel; novel therapeutics; photoreceptor degeneration; preservation; promoter; repaired; response; retinal rods; therapeutic genome editing; therapeutic target; therapeutically effective; tool; vector

PROJECT SUMMARY Retinitis pigmentosa (RP) is the most common inherited retinal dystrophy (IRD), caused by more than 3,100 mutations in 80 genes that are primarily specific to rod photoreceptors. Following major rod death phase, cone death occurs regardless of the underlying gene mutations. Currently, there exists a knowledge gap in understanding how aerobic glycolysis in photoreceptors impact the delicate “metabolic coupling” between rods and the retinal pigment epithelium (RPE) in RP. The long-term goal of this project is to develop a therapy that will preserve cone function in patients with RP. The objectives of this proposal are to investigate how metabolic dysregulation due to lactate deficiency contributes to photoreceptor death in RP, test a novel metabolome reprogramming strategy, and fulfill important safety requirements for filing a Pre-Investigational New Drug application. The hypothesis is that reprogramming rod and cone aerobic glycolysis will promote cone survival in RP independent of the underlying rod-specific gene mutations. This hypothesis has been formulated based on the applicant’s strong preliminary data. The rationale for the proposed research is that by targeting a metabolic pathway common to many of the genetically heterogeneous forms of RP, cone function may be preserved for the equivalent of 10 or more human years, which would have a tremendously positive impact on the lives of patients with RP. This hypothesis will be tested by pursuing three specific aims: 1) Investigate whether photoreceptor-specific ablation of prolyl hydroxylase domain-containing protein (Phd), a metabolic enzyme that inhibits aerobic glycolysis under normoxia, preserves cones by enhancing aerobic glycolysis in an autosomal recessive RP mouse model; 2) Assess the efficacy and feasibility of enhancing cone survival and function by ablating PHD2 in photoreceptors of a dominant RP mouse model ; 3) Establish the safety of therapeutic Phd2 editing in a WT and RP mouse model as well as human cells. Specifically, Aim 1 will determine whether enhanced aerobic glycolysis in cone photoreceptors can promote their survival in a novel genetic mouse model. Aim 2 will test the potential of gene therapy to slow photoreceptor degeneration by enhancing aerobic glycolysis in a different mouse model of RP. Lastly, Aim 3 will define the pharmacokinetics and safety of the aerobic glycolysis reprogramming vector. The approach is innovative because this will be the first example of cell-specific CRISPR-mediated precision metabolic reprogramming and because the novel therapeutic-editing vectors can be redeployed in future Phase I-IIA trials without modification. The proposed research is significant as it has the potential to dramatically lower the cost of treatment, be applicable to dividing and nondividing cells, shape ongoing CRISPR research, and ultimately define aerobic glycolysis as a safe and effective therapeutic target.

项目摘要 色素性视网膜炎（RP）是最常见的视网膜营养不良（IRD），由3100多个 主要针对杆感光体的80个基因中的突变。在大杆死亡阶段，锥 死亡都是不论基本基因突变的。目前，存在一个知识差距 了解光感受器中的有氧糖酵解如何影响杆之间细腻的“代谢耦合” RP中的视网膜色素上皮（RPE）。该项目的长期目标是开发一种疗法 RP患者将保留锥功能。该提案的目标是调查新陈代谢 由于裂缝缺乏引起的失调导致RP感光受体死亡，测试一种新型代谢组 重新编程策略，并满足提交预先投票的新药的重要安全要求 应用。假设是重编程杆和有氧糖酵解将促进锥体存活 RP独立于潜在的杆特异性基因突变。该假设已根据 申请人的强大初步数据。拟议研究的理由是，针对代谢 许多一般异构形式的RP共有的途径，可以保留锥函数 相当于10个或以上的人类年份，这将对的生活产生巨大的积极影响 RP患者。该假设将通过追求三个具体目的来检验：1）调查是否研究 含丙酰羟化酶结构蛋白（PHD）的光感受器特异性消融，一种代谢酶 抑制有氧糖酵解在常氧中，通过在常染色体中增强有氧糖酵解来保存锥 隐性RP鼠标模型； 2）评估增强锥体存活和功能的效率和可行性 在主要的RP小鼠模型的光感受器中烧蚀PHD2； 3）建立治疗性PHD2的安全性 在WT和RP小鼠模型以及人类细胞中进行编辑。具体来说，AIM 1将确定是否 锥形感光体中增强的有氧糖酵解可以在新型的遗传小鼠模型中促进其生存。 AIM 2将通过增强有氧糖酵解来测试基因疗法对降低感光受体变性的潜力 在不同的RP鼠标模型中。最后，AIM 3将定义有氧运动的药代动力学和安全性 糖酵解重编程载体。该方法具有创新性，因为这将是细胞特异性的第一个例子 CRISPR介导的精确代谢重编程，因为新型治疗载体可以 在未来的I-IIA试验中重新部署，而无需修改。拟议的研究很重要，因为它具有 明显降低治疗成本的潜力，适用于分裂和非分散细胞，形状 正在进行的CRISPR研究，并最终将有氧糖酵解定义为安全有效的治疗靶点。