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

基本信息

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
项目类别：
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

项目摘要

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希望。然而它的应用在由于缺乏安全的分娩工具以及所需的大量成本和时间投入，诊所进展缓慢定制开发个体疗法。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核糖核蛋白 (cRNP) 的基因传递系统，专门针对神经元细胞鞘内（IT）给药后遍及大脑。该应用程序的总体目标是应用此用于治疗两种严重神经发育障碍（NDD）的新型基因编辑技术： Angelman 综合征 (AS) 和 H1-4 综合征（HIST1H1E 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记UBE3A基因缺陷引起的疾病染色体 15q11-q13 区域。在所有 AS 病例中，父本染色体中的 UBE3A 结构都是完整的但转录受到 UBE3A (UBE3A-ATS) 介导的非编码和反义 RNA 的抑制机制。已经令人信服地证明，通过反义寡核苷酸 (ASO) 还原 UBE3A-ATS，拓扑异构酶抑制剂和病毒传递的 Cas9 基因编辑可以去抑制 UBE3A 和纠正 AS 小鼠模型中异常的神经表型。 H1-4 综合征是由于获得功能机制归因于具有异常 C 末端移码尾 (CFT) 的突变蛋白。 H1-4 综合征有与 AS 相似但较轻的临床特征。 H1-4综合征尚无有效的干预措施。因此，一个长期的迫切需要针对 AS 和 H1-4 以及其他 NDD 的长期分子治疗。在我们的临床前研究中使用一个经过充分验证的 AS 小鼠模型，我们证明了 Ube3a 反义靶向的单次 IT 交付 RNP/Cas9有效地去抑制父本染色体上Ube3a的表达，从而导致校正神经行为表型。同样，H1-4 CFT 的敲除可以挽救 H1-4 中的异常表型。 4只人源化小鼠。我们提出基于 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）、治疗机制（基因失活）和总体试验设计。我们组建了一支来自耶鲁大学的优秀团队和拉什大学在临床前、IND研究领域拥有强大且互补的专业知识，和临床试验。这项研究的成功将带来有史以来第一个基于基因编辑的 AS 和 H1-治疗方法 4.更重要的是，它将支持基因组编辑的范式转变；数量迅速扩大体内基因编辑治疗神经遗传疾病并加速基因组编辑转型技术进入临床应用。