Screening for rAAV transduction enhancers

筛选 rAAV 转导增强子

• 批准号: 9255016
• 负责人: Terry L. Bowlin
• 金额: $ 30万
• 依托单位: MICROBIOTIX, INC
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-01-27 至 2018-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9255016
• 关键词: Alkaline Phosphatase; Alpha Cell; Animal Model; Antibodies; Binding; Biological Assay; CCR5 gene; CD8-Positive T-Lymphocytes; Capsid; Cell Culture Techniques; Cell Line; Cell Nucleus; Cells; Complementary DNA; Dengue Infection; Detection; Development; Dose; Drug Design; Embryo; Engineering; Enhancers; Fluorescence; Funding; Gene Delivery; Gene Transduction Agent; Genome; Glycoproteins; Goals; Grant; HIV; HIV-1; Human; Immune response; In Vitro; Infection; Influenza; Intramuscular; Laboratories; Lead; Libraries; Ligands; Macaca; Measurement; Measures; Mediating; Nuclear; Phase; Production; Prophylactic treatment; Proteins; Public Health; Recombinant adeno-associated virus (rAAV); Reporter; Reporter Genes; Research Institute; Research Proposals; Role; Route; SIV; Safety; Shapes; Signal Transduction; TLR9 gene; Therapeutic; Time; Transgenes; Treatment Cost; Vaccine Therapy; Vaccines; Viral; Viral Genome; Virus; adaptive immune response; base; cost; cytotoxic; design; ds-DNA; gene therapy; high throughput screening; malaria infection; mimetics; multicatalytic endopeptidase complex; neutralizing antibody; particle; phase 2 study; programs; protein expression; response; scaffold; screening; small molecule; trafficking; transgene expression; vector; virus envelope

ABSTRACT Recombinant adeno-associated virus (rAAV) is a promising gene delivery vector for therapeutic and vaccine applications, but its low transduction efficacy currently limits its utility. The primary reasons for low transduction efficacy are as follows: (i) inefficient trafficking of the rAAV genome to the nucleus, (ii) inefficient host cell-mediated synthesis of double-stranded DNA from the single-stranded genome, and (iii) Toll like receptor (TLR) 9 responses. The goal of this proposal is to identify small molecules that increase the transduction efficacy of rAAV transduction by modulating any one of the barriers. One immediate application will be the use of these compounds in the rAAV-based gene-therapy against HIV developed in Dr. Michael Farzan’s laboratory at The Scripps Research Institute. The rAAV vector expresses an antibody-like immunoadhesin (eCD4-Ig), a fusion of CD4-Ig with a small CCR5-mimetic sulfopeptide, which binds to the HIV-1/simian immunodeficiency virus (SIV) envelope glycoprotein and protects macaques from infection. Increased transduction efficacy will decrease both the cost of treatment and the adaptive immune response to the viral capsid and expressed transgene that limits the use of rAAV vectors in humans. However, the significance of our studies goes beyond HIV-1 prophylaxis. These small molecules will be useful for other rAAV-based vaccines and therapeutics, such as vaccines developed for influenza, malaria, and dengue infection, or for the long-term delivery of biologics. In Phase I, we will (i) develop a high-throughput screening (HTS) assay to identify compounds that enhance rAAV transduction, (ii) identify transduction enhancers, (iii) broadly evaluate the mode of action, and prioritize hits for a Phase II study. Promising scaffolds will be subjected to a rational drug design program in Phase II, and will be advanced to IND enabling studies.

抽象的 重组腺相关病毒（RAAV）是一种有前途的基因递送载体 和疫苗应用，但其低转移效率目前限制了其效用。主要 低转移效率的原因如下：（i）RAAV基因组效率低下 到核，（ii）宿主细胞介导的双链DNA的效率低下的宿主细胞介导的合成 单链基因组和（iii）Toll像受体（TLR）9反应。该提议的目标 是确定小分子，以提高RAAV转导的转移效率 调节任何一个障碍。一种即时应用是使用这些 基于RAAV的基因治疗中针对HIV的化合物在Michael Farzan博士的 斯克里普斯研究所的实验室。 RAAV载体表达类似抗体的 免疫粘附素（ECD4-Ig），CD4-Ig与小CCR5模拟硫化物的融合 结合HIV-1/Simian免疫缺陷病毒（SIV）包膜糖蛋白并保护 感染的猕猴。提高的转移效率将降低 治疗和对病毒衣壳的适应性免疫反应，并表达转基因 限制在人类中使用Raav载体。但是，我们的研究的意义超出了 HIV-1预防。这些小分子将对其他基于RAAV的疫苗和 治疗剂，例如为影响力，疟疾和登革热感染而开发的疫苗，或 生物制剂的长期交付。在第一阶段，我们将（i）开发高通量筛选（HTS） 测定确定增强RAAV转导的化合物的测定，（ii）识别转导 增强剂（iii）广泛评估行动方式，并优先考虑II期研究的命中。 有希望的脚手架将在第二阶段进行合理的药物设计计划，并将 先进的研究促进研究。