A non-viral CRISPR-mediated genome editing delivery platform as a potential therapy for neurogenetic diseases

非病毒 CRISPR 介导的基因组编辑传递平台作为神经遗传疾病的潜在疗法

• 批准号: 10739113
• 负责人: Elizabeth Mara Berry-Kravis
• 金额: $ 501.9万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-20 至 2026-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10739113
• 关键词: 17 year old; Acceleration; Adopted; Adult; Angelman Syndrome; Animals; Antisense Oligonucleotides; Antisense RNA; Area; Biodistribution; Brain; C-terminal; CRISPR/Cas technology; Cells; Chemicals; Child; Chromosomes; Clinic; Clinical; Clinical Treatment; Clinical Trials; Clinical Trials Design; Clustered Regularly Interspaced Short Palindromic Repeats; Communities; Complex; Contracts; Custom; Data; Development; Disease; Dose; Drug Kinetics; Enrollment; Event; Family; Foundations; Funding; Future; Genes; Genetic; Genetic Diseases; Genome; Genomics; Goals; Guide RNA; Human; Human Chromosomes; Impairment; Individual; Infrastructure; Intrathecal Injections; Investments; Manuals; Mediating; Modeling; Modification; Molecular; Monkeys; Mus; Mutate; Neurodevelopmental Disorder; Neurologic; Neurons; Organoids; Outcome Measure; Patient Recruitments; Patients; Penetration; Perinatal mortality demographics; Pharmacology Study; Pharmacology and Toxicology; Phase; Phase I Clinical Trials; Phenotype; Production; Proteins; Protocols documentation; Rahman Syndrome; Regulatory Affairs; Research; Ribonucleoproteins; Rodent; Route; Safety; Signal Transduction; Structure; Surface; Syndrome; System; Tail; Technology; Therapeutic; Time; Topoisomerase Inhibitors; Toxicology; UBE3A gene; United States National Institutes of Health; Universities; Untranslated RNA; Virus; Work; adverse event monitoring; base editing; behavioral study; cell type; clinical application; cost; derepression; design; effective intervention; experience; first-in-human; gain of function; gene delivery system; gene repression; genome editing; humanized mouse; immunogenicity; imprint; improved; in vivo; induced pluripotent stem cell; knock-down; mouse model; neurobehavioral; neurogenetics; novel; novel strategies; patient retention; pediatric patients; phase 1 study; pre-Investigational New Drug meeting; pre-clinical; preclinical study; primary outcome; programs; rare genetic disorder; response; safety assessment; somatic cell gene editing; success; tool; trial design

Genome editing holds great promise for the treatment of many genetic diseases; however its application in the clinic has been slow due to the lack of the safe delivery tools and significant cost and time investment required to custom-develop individual therapies. In our SCGE program phase 1 study, we developed a chemically modified ribonucleoprotein (cRNP)-based gene delivery system that specifically targets neuronal cells throughout the brain after intrathecal (IT) administration. The overarching goal of this application is to apply this novel gene editing technology towards the treatment of two severe neurodevelopmental disorders (NDD): Angelman syndrome (AS) and H1-4 syndrome (HIST1H1E syndrome). AS is a devastating neurodevelopmental disease caused by the deficiency of the maternal and brain specific imprinting UBE3A gene in human chromosome 15q11-q13 region. The structure of UBE3A is intact in the paternal chromosome in all AS cases but transcriptionally repressed by a non-coding and antisense RNA of UBE3A (UBE3A-ATS) mediated mechanism. It has been shown convincingly that reduction of UBE3A-ATS by antisense oligo (ASO), topoisomerase inhibitors, and virus delivered Cas9 gene editing can de-repress the expression of UBE3A and correct the abnormal neurological phenotypes in AS mouse models. H1-4 syndrome is caused by a gain of function mechanism due to a mutated protein with aberrant C-terminal frameshift tail (CFT). H1-4 syndrome has similar but milder clinical features than AS. There is no effective intervention for H1-4 syndrome. Thus, a long- term molecular therapy for AS and H1-4, as well as other NDDs is urgently needed. In our preclinical study using a well validated AS mouse model, we demonstrated that a single IT delivery of Ube3a antisense-targeting RNP/Cas9efficiently de-represses the expression of Ube3a from the paternal chromosome, leading to correction of neurobehavioral phenotypes. Similarly, the knockdown of H1-4 CFT rescue the abnormal phenotypes in H1- 4 humanized mice. We propose our cRNP-based platform for the treatment of AS and H1-4 syndrome utilizing the same genome editor (CRISPR), delivery system (cRNPs), route (IT), target cell type (neurons), therapeutic mechanism (genetic inactivation) and overall trial design. We have assembled an outstanding team from Yale and Rush University with strong and complementary expertise in the areas of preclinical, IND enabling studies, and clinical trials. The success of this study will lead to the first ever gene editing based therapy for AS and H1- 4. More importantly, it will support a paradigm shift for genome editing; rapidly expanding the number of neurogenetic diseases treated by in vivo gene editing and accelerating the transition of genome editing technology into clinical applications.

基因组编辑为治疗许多遗传疾病带来了巨大的希望;然而，它在遗传疾病中的应用 由于缺乏安全的分娩工具，需要投入大量的费用和时间，诊所的工作进展缓慢 定制开发个性化疗法。在我们的SCGE项目第一阶段研究中，我们开发了一种化学方法， 基于修饰的核糖核蛋白（cRNP）的基因递送系统，其特异性靶向神经元细胞 在鞘内（IT）给药后在整个脑中。此应用程序的首要目标是应用此 新的基因编辑技术用于治疗两种严重的神经发育障碍（NDD）： Angelman综合征（AS）和H1-4综合征（HIST 1H 1 E综合征）。AS是一种破坏性的神经发育疾病， 人类母体和大脑特异性印迹UBE 3A基因缺陷引起的疾病 染色体15 q11-q13区域。在所有AS病例中，UBE 3A在父亲染色体中的结构是完整的 但被UBE 3A的非编码和反义RNA（UBE 3A-ATS）介导的转录抑制， 机制已经令人信服地表明，通过反义寡核苷酸（阿索）减少UBE 3A-ATS， 拓扑异构酶抑制剂和病毒递送的Cas9基因编辑可以去抑制UBE 3A的表达， 纠正AS小鼠模型的异常神经表型。H1-4综合征是由获得 由于具有异常C-末端移码尾（CFT）的突变蛋白质而导致的功能机制。H1-4综合征 临床特征与AS相似但较轻。H1-4综合征尚无有效的干预措施。因此，一个长期的- 目前，AS和H1-4以及其他NDD的长期分子治疗是迫切需要的。在我们的临床前研究中， 一个经过充分验证的AS小鼠模型，我们证明了Ube 3a反义靶向的单次IT递送 RNP/Cas9有效地去抑制来自父本染色体的Ube 3a的表达，导致校正 神经行为表型类似地，H1-4 CFT的敲低挽救了H1- 4中的异常表型。 4只人源化小鼠。我们提出了基于cRNP的平台，用于治疗AS和H1-4综合征， 相同的基因组编辑器（CRISPR）、递送系统（cRNP）、途径（IT）、靶细胞类型（神经元）、治疗性 机制（基因失活）和总体试验设计。我们召集了一支来自耶鲁的优秀团队 和拉什大学在临床前，IND使能研究领域拥有强大的互补专业知识， 和临床试验。这项研究的成功将导致有史以来第一个基于基因编辑的AS和H1治疗。 4.更重要的是，它将支持基因组编辑的范式转变;快速扩大基因组编辑的数量。 通过体内基因编辑和加速基因组编辑的转变来治疗神经遗传性疾病 技术应用于临床。