Dissecting AAV silencing in humanized mice

解析人源化小鼠中的 AAV 沉默

基本信息

批准号：
10562376
负责人：
Aravind Asokan
金额：
$ 61.62万
依托单位：
DUKE 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：
2023-02-01 至 2027-01-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10562376
关键词：
Acute Advisory Committees Animal Model Binding Biodistribution Biological Assay Biology CRISPR-mediated transcriptional activation Capsid Cells Clinical Clinical Trials Complex Data Deposition Development Disease Dose Engineering Enzymes Epigenetic Process Event Failure Gene Expression Gene Silencing Genetic Transcription Genome Genome Components Genomic DNA Glycogen storage disease type II Hemophilia A Hemophilia B Hepatocyte Hepatocyte transplantation Hepatolenticular Degeneration Hepatotoxicity Histone Deacetylase Inhibitor Histones Human Immune response Implant In Vitro Individual Integration Host Factors Knockout Mice Lentivirus Vector Liver Liver diseases Maps Mediating Metabolic Methylation Modeling Molecular Mouse Strains Mus Organ Ornithine carbamoyltransferase deficiency Patients Pattern Pre-Clinical Model Process Promoter Regions Reporting Repression Role Serotyping Serum Steroids T cell response Therapeutic Transgenes Tropism Urea cycle disorders Variant Viral Viral Genes Viral Genome Viral Vector Virus Receptors Work Xenograft procedure adeno-associated viral vector canine model clinical translation clinically relevant derepression design experience gene product gene therapy gene therapy clinical trial genome editing genotoxicity human disease humanized mouse in vivo innovation knock-down meetings methylmalonic aciduria mouse model nonhuman primate novel pharmacologic prevent promoter response restoration small hairpin RNA therapeutic target therapeutic transgene transcription factor transgene expression uptake vector vector genome viral genomics

项目摘要

PROJECT SUMMARY The liver has emerged as a promising target for expressing therapeutic transgenes utilizing Adeno-Associated Viral (AAV) vector-mediated delivery. Despite numerous clinical trials and continued progress several challenges have been identified for AAV gene therapy. In this proposal we will dissect a major clinical hurdle, e.g. AAV transgene silencing in the human liver and molecular mechanisms underlying this phenomenon. To achieve such, we first propose to utilize and characterize a novel humanized liver model, which will allow precise and selective interrogation of the mechanisms underlying AAV transgene silencing in normal as well as diseased human hepatocytes in vivo. The second aim will build on a recent observation from our lab showing AAV transgene expression mediated by the Human Silencing Hub (HUSH) complex. We will further dissect and gather mechanistic inside on this process using human hepatocytes from methylmalonic acidemia (MMA) patients in vivo. Hence MMA will serve as a proof of concept disorder for liver directed AAV gene therapy. Finally, we will explore different vector design and pharmacological strategies to rescue AAV gene silencing.

项目概要肝脏已成为表达治疗性转基因的有希望的靶标利用腺相关病毒（AAV）载体介导的递送。尽管进行了大量的临床试验并持续取得进展 AAV 基因治疗面临一些挑战。在这个我们将剖析一个主要的临床障碍，例如人肝脏中 AAV 转基因沉默以及这一现象背后的分子机制。为了实现这一目标，我们首先建议利用并表征新型人源化肝脏模型，这将允许精确和选择性探究正常和正常情况下 AAV 转基因沉默的机制体内患病的人肝细胞。第二个目标将建立在最近的观察基础上我们的实验室展示了由人类沉默中心 (HUSH) 介导的 AAV 转基因表达复杂的。我们将利用人类进一步剖析和收集这一过程的内部机制。来自甲基丙二酸血症（MMA）患者的体内肝细胞。因此 MMA 将作为肝脏定向 AAV 基因治疗的概念验证障碍。最后，我们将探索不同的拯救 AAV 基因沉默的载体设计和药理学策略。