Engineered AAV vectors for combinatorial treatment of rare genetic brain diseases

用于罕见遗传性脑部疾病组合治疗的工程 AAV 载体

• 批准号: 10414342
• 负责人: VIRGINIA Eunice KIMONIS
• 金额: $ 60万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10414342
• 关键词: Engineered; AAV; vectors; combinatorial; treatment

Project Summary AAV-based gene therapy requires the development of safe, efficient, and target-specific vectors. AAV-mediated gene therapy for peripheral tissues (blood and skeletal muscle) has made great strides, focused mainly on gene replacement for loss of function diseases. However, there has been little research on dominantly inherited CNS disorders that are caused by toxic gene products. We have assembled a world class, multi-disciplinary academic research team supported by our industrial partners to develop innovative AAV-based gene knockdown and replacement treatments for rare genetic diseases including Spinocerebellar Ataxia Type 7 (SCA7) and valosin- containing protein (VCP) multisystem proteinopathy. We respond to the RFA Project Objectives and propose three Specific Aims. In Aim 1, we will design and manufacture new AAV vectors with improved critical quality attributes (safety, efficacy, target specificity) for gene therapy. We propose innovative neuron specific gene delivery, temporal control of gene expression and reduced immune responses in the CNS. We have constructed AAV vectors that express EGFP and mRNA barcodes for improved screening to support our proposed AAV treatments. In Aim 2, we will develop advanced quantitative analytics by combining next-generation sequencing and bar-coded AAVs for efficient assessment of in vivo gene delivery in the mouse model. We will screen and compare AAV-mediated gene expression with different capsid variants using different promoters / enhancers. Single-cell RNAseq also will be used to assess immunological responses of selected AAV vectors by different administration routes. In Aim 3, to effectively treat the SCA7 and VCP diseases, we will develop novel AAV vectors that simultaneously knockdown toxic gene products while replacing normal gene products that are codon optimized to be unaffected by knockdown. This proposed combinatorial treatment will establish a proof-of- concept for many other dominant inherited diseases, where loss of normal allele expression due to non-specific silencing causes its own problems. We have well developed models of the SCA7 and VCP diseases for Aim 3 studies. Our team has great expertise in developing, manufacturing and applying AAV vectors in basic research and preclinical application. Our published work and preliminary data establish the feasibility and key methodologies for the proposed research. Critically, we have an established viral production facility and distribution platform operating out of our UCI Center for Neural Circuit Mapping that supports viral reagent design, validation, and manufacturing.

项目摘要 基于AAV的基因治疗需要开发安全、高效和靶向特异性的载体。aav介 外周组织（血液和骨骼肌）的基因治疗已经取得了长足的进步，主要集中在基因治疗方面。 替代功能丧失的疾病。然而，对显性遗传CNS的研究很少 由有毒基因产物引起的疾病我们聚集了一个世界级的，多学科的学术 在我们的工业合作伙伴的支持下，我们的研究团队开发了创新的基于AAV的基因敲除， 罕见遗传疾病的替代治疗，包括脊髓小脑共济失调7型（SCA 7）和valosin- 多系统蛋白质病（VCP）我们响应RFA项目目标，并建议 三个具体目标。在目标1中，我们将设计和制造具有改进的关键质量的新的AAV载体 基因治疗的属性（安全性、有效性、靶点特异性）。我们提出了创新的神经元特异性基因 递送、基因表达的时间控制和CNS中免疫应答的降低。我们已经构建 表达EGFP和mRNA条形码的AAV载体，用于改进筛选以支持我们提出的AAV 治疗。在目标2中，我们将通过结合下一代测序技术， 和条形码化的AAV用于在小鼠模型中有效评估体内基因递送。我们会筛选， 比较使用不同启动子/增强子的具有不同衣壳变体的AAV介导的基因表达。 单细胞RNAseq还将用于评估所选AAV载体通过不同的免疫应答。 给药途径。在目标3中，为了有效地治疗SCA 7和VCP疾病，我们将开发新的AAV， 同时敲低毒性基因产物同时替换正常基因产物的载体， 优化为不受击倒的影响。这种组合疗法将建立一个证明- 许多其他显性遗传性疾病的概念，其中由于非特异性基因突变导致正常等位基因表达的丧失， 沉默本身就有问题。我们已经为Aim 3开发了SCA 7和VCP疾病的良好模型 问题研究我们的团队在基础研究中开发，制造和应用AAV载体方面具有丰富的专业知识 和临床前应用。我们已发表的工作和初步数据建立了可行性和关键 建议的研究方法。关键是，我们有一个成熟的病毒生产设施， 在我们的UCI神经回路映射中心外运行的分销平台，支持病毒试剂设计， 验证和制造。