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.

摘要