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

基本信息

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
项目类别：
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&apos s Disease Alzheimer&apos 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

项目摘要

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件递送至大脑以治疗神经退行性疾病抽象的 CRISPR 技术作为精确基因编辑的变革工具包可能会解决许多棘手的神经退行性疾病然而，我们之前将基于 CRISPR 的基因编辑元件传递到大脑的效率非常低。率先使用聚焦超声 (FUS) 技术来实现无创、针对大脑的递送。证明 FUS 与单分散、充气微泡结合可以提供治疗有效负载，包括小分子、蛋白质和腺相关病毒（AAV），通过静脉注射到达大脑的特定区域此外，我们还重新设计了 Polyplex 系统。提高非病毒 CRISPR 介导的基因编辑在体外和体内的效率和特异性。初步研究中，我们建议整合组织特异性（FUS）和细胞特异性（AAV 和非病毒）的优点 Polyplex 载体）递送系统，使 CRISPR 能够递送至大脑及其疾病相关细胞类型。该项目的目标是开发一种非侵入性、FUS 介导的技术，用于传递 AAV 载体和非病毒将 CRISPR 元件携带到大脑的多聚体，并评估对两种主要神经退行性疾病的功效，我们建议在 UG3 发展阶段追求三个具体目标和一个目标。 UH3 演示阶段的第四个目标：(1) 通过确定的微泡成分、声学特性来优化 FUS 系统参数并优化携带 CRISPR 敲除、抑制以及激活元件的 AAV 载体 (2) 开发高效的 CRISPR 递送非病毒复合物系统 (3) 疗效评价及疗效评价通过优化的 AAV 和非病毒 CRISPR 传递系统，确保阿尔茨海默病和帕金森病模型的安全性； (4)评估优化后的递送系统在NHP模型中的递送效率和安全性。仅依靠颅内注射将药物输送到大脑，该项目将是第一个开发该项目的成功将激发新的战略。治疗仍然具有挑战性甚至无法治愈的神经退行性疾病的方法。