Focused Ultrasound-mediated Delivery of Gene-editing Elements to the Brain for Neurodegenerative Disorders

聚焦超声介导的基因编辑元件递送至大脑以治疗神经退行性疾病

• 批准号: 9810901
• 负责人: KAM W LEONG
• 金额: $ 79.85万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-01 至 2022-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9810901
• 关键词: Acoustics; Adverse effects; Alzheimer' s Disease; Alzheimer' s disease model; Amyloid beta-Protein; Amyloid beta-Protein Precursor; Astrocytes; Brain; Brain region; CRISPR/Cas technology; Cells; Clustered Regularly Interspaced Short Palindromic Repeats; Dependovirus; Development; Disease; Disease model; Elements; Expression Profiling; Focused Ultrasound; Gases; Gene Delivery; Generations; Genes; In Situ; In Vitro; Injections; Knock-out; Length; Mediating; Methodology; Microbubbles; Minor; Modeling; Mus; Neurodegenerative Disorders; Neurons; Parkinson Disease; Pathway interactions; Phase; Phenotype; Physiologic pulse; Plasmids; Play; Pluronics; Proteins; Proteolysis; Reporting; Rodent; Safety; Senile Plaques; Specificity; Structure; System; Techniques; Technology; Testing; Therapeutic; Tissues; Transcription Coactivator; Transcription Repressor/Corepressor; Treatment Efficacy; Viral Genome; Viral Packaging; Virion; adeno-associated viral vector; aged; base; beta-site APP cleaving enzyme 1; cell type; design; dopaminergic neuron; dosage; gene delivery system; immunogenicity; improved; improved outcome; in vivo; intravenous injection; mutant; non-viral gene delivery; nonhuman primate; novel therapeutic intervention; pressure; prevent; small molecule; success; therapeutic gene; transcription factor; vector

Focused Ultrasound-mediated Delivery of Gene-editing Elements to the Brain for Neurodegenerative Disorders Abstract CRISPR technology as a transformative toolkit for precise gene editing may tackle many intractable neurodegenerative diseases. However, delivery of CRISPR-based gene editing elements to the brain is highly inefficient. We previously pioneered the use of focused ultrasound (FUS) technology to achieve noninvasive, brain-specific delivery. We demonstrated that FUS in conjunction with monodispersed, gas-filled microbubbles could deliver therapeutic payloads, including small molecules, proteins and adeno-associated virus (AAV), to a specific region of the brain through intravenous injection in both rodent and non-human primate (NHP) models. Furthermore, we have also redesigned polyplex systems to improve the nonviral CRISPR-mediated gene editing efficiency and specificity both in vitro and in vivo. With those preliminary studies, we propose to integrate the merits of tissue-specific (FUS) and cell-specific (AAV and non-viral polyplex vectors) delivery systems to enable CRISPR delivery to the brain and its disease-related cell types. The overall objective of this project is to develop a non-invasive, FUS-mediated technology for delivering AAV vectors and non-viral polyplexes carrying CRISPR elements to the brain and to evaluate the efficacy on two major neurodegenerative disorders, Alzheimer’s and Parkinson’s diseases. We propose to pursue three specific aims in the UG3 developmental phase and a fourth aim in the UH3 demonstration phase: (1) Optimize the FUS system with defined microbubble composition, acoustic parameters and optimize the AAV vector carrying the CRISPR knockout, suppression as well as activation elements to achieve high gene editing efficacy in mouse brain; (2) Develop an efficient CRISPR delivery non-viral polyplex system with a transient expression profile for delivery into the brain via FUS technology; (3) Evaluate the therapeutic efficacy and the safety on both Alzheimer’s and Parkinson’s disease models with the optimized AAV and non-viral CRISPR delivery systems; and (4) Evaluate the delivery efficiency and safety of the optimized delivery system in the NHP model. As current CRISPR delivery to the brain relies only on intracranial injection, the proposed project will be the first study developing a noninvasive, efficient approach to achieve gene editing in the brain. Success of this project will stimulate new strategic approaches of tacking neurodegenerative disorders that remain challenging or even untreatable.

聚焦超声介导的基因编辑元素向大脑的递送，用于神经退行性疾病 抽象的 CRISPR技术作为用于精确基因编辑的变革性工具包，可能会解决许多棘手的神经退行性 疾病。但是，将基于CRISPR的基因编辑元件传递到大脑的效率高度低。我们以前 开创了使用专注的超声（FUS）技术来实现非侵入性，大脑特异性的传递。我们 证明FUS与单分散，充满气体的微泡结合使用可以提供热电载荷， 包括小分子，蛋白质和腺相关病毒（AAV），通过静脉注射到大脑的特定区域 注射啮齿动物和非人类灵长类动物（NHP）模型。此外，我们还重新设计了多链体系统 为了提高体外和体内的非病毒CRIS介导的基因编辑效率和特异性。与那些 初步研究，我们建议整合组织特异性（FUS）和细胞特异性的优点（AAV和非病毒 息肉矢量）输送系统，可以使CRISPR递送到大脑及其与疾病相关的细胞类型。总体 该项目的目的是开发一种非侵入性的FUS介导的技术，用于提供AAV媒介和非病毒 将CRISPR元素带到大脑的双流线物质，并评估两个主要神经退行性疾病的效率，即 阿尔茨海默氏症和帕金森氏病。我们建议在UG3发展阶段实现三个特定目标，并 在UH3演示阶段的第四个目标：（1）用定义的微泡组成优化FUS系统 参数并优化携带CRISPR敲除，抑制以及激活元素的AAV矢量 在小鼠脑中实现高基因编辑效率； （2）与 通过FUS技术传递到大脑的瞬态表达曲线； （3）评估治疗效率和 具有优化的AAV和非病毒CRISPR递送系统的阿尔茨海默氏病和帕金森氏病模型的安全性； （4）评估NHP模型中优化输送系统的交付效率和安全性。作为当前的CRISPR 向大脑的交付仅依赖颅内注射，拟议的项目将是第一个开发的研究 在大脑中实现基因编辑的无创，有效的方法。该项目的成功将刺激新策略 处理仍然受到挑战甚至无法治疗的神经退行性疾病的方法。