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.

基因组编辑对许多遗传疾病的治疗有很大的希望。但是它在由于缺乏安全的交付工具以及所需的大量成本和时间投资，诊所一直很慢定制开发个人疗法。在我们的SCGE计划1阶段研究中，我们开发了一个化学的基于改性的核糖核蛋白（CRNP）基因递送系统，该系统专门针对神经元细胞鞘内（IT）给药后整个大脑。此应用程序的总体目标是应用此新型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（Hist1H1E综合征）。正如毁灭性的神经发育由人类和大脑特异性印记的Ube3a基因缺乏引起的疾病染色体15q11-Q13区域。在所有情况下但是通过ube3a（ube3a-ats）介导的非编码和反义RNA的转录抑制机制。令人信服地表明，反义寡素（ASO）减少UBE3A-ATS，拓扑异构酶抑制剂和传递CAS9基因编辑的病毒可以抑制UBE3A的表达和纠正AS模型中的异常神经表型。 H1-4综合征是由增益引起的由于突变的蛋白质引起的功能机制，具有异常的C末端移码尾（CFT）。 H1-4综合征具有类似但临床温度较温和的特征。 H1-4综合征没有有效的干预。因此，长期迫切需要AS和H1-4以及其他NDD的术语分子疗法。在我们使用的临床前研究中一个经过良好验证为鼠标模型的，我们证明了UBE3A反义靶向的单个IT输送 RNP/cas9有效地脱氧于父亲染色体的表达，导致校正神经行为表型。同样，H1-4 CFT的敲低挽救了H1-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综合征同一基因组编辑器（CRISPR），输送系统（CRNP），路线（IT），靶细胞类型（神经元），治疗性机理（遗传失活）和整体试验设计。我们已经组建了一支来自耶鲁大学的杰出团队以及在临床前，有能力研究领域的强大和互补专业知识的Rush University，和临床试验。这项研究的成功将导致AS和H1-的第一个基于基因编辑的疗法 4。更重要的是，它将支持基因组编辑的范式转变；迅速扩大数量通过体内基因编辑治疗和加速基因组编辑过渡的神经遗传疾病技术进入临床应用。