RNA-mediated excision of the HIV-1 genome from latently infected cells in CNS

RNA介导的中枢神经系统潜伏感染细胞中HIV-1基因组的切除

• 批准号: 8995716
• 负责人: Wenhui Hu
• 金额: $ 39万
• 依托单位: TEMPLE UNIV OF THE COMMONWEALTH
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-03-15 至 2017-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8995716
• 关键词: Ablation; Acquired Immunodeficiency Syndrome; Adenoviruses; Aging; Aging-Related Process; Anti-Retroviral Agents; Astrocytes; Bioinformatics; Biological Assay; Biological Models; Biology; Bone Diseases; Brain; Cell Culture Techniques; Cell model; Cells; Centers for Disease Control and Prevention (U.S.); Central Nervous System Diseases; Chromosomes; Clinical Research; Comorbidity; Complementary RNA; DNA; DNA Sequence; Detection; Development; Dideoxy Chain Termination DNA Sequencing; Disease; Drug resistance; Employment; Enzyme-Linked Immunosorbent Assay; Epidemic; Excision; Exhibits; Gene Expression; Gene Mutation; Gene Targeting; Genes; Genetic Transcription; Genome; Genotype; Guide RNA; HIV-1; Heart failure; Hela Cells; Human; In Vitro; Individual; Infection; Knowledge; Laboratories; Lentivirus Vector; Long-Term Survivors; Luciferases; Mediating; Microglia; Modeling; Molecular; Molecular Genetics; Neuropathogenesis; Outcome; Outcome Study; Patients; Pharmaceutical Preparations; Phenotype; Population; Production; Property; Proteins; Quality of life; RNA; Reporter; Reporter Genes; Reporting; Rest; Restriction fragment length polymorphism; Risk; Solid; Southern Blotting; Specificity; Survival Rate; System; Technology; Therapeutic; Uncertainty; United States; Viral; Viral Genes; Viral Genome; Viral Proteins; Viral reservoir; Virus; Virus Diseases; Virus Replication; antiretroviral therapy; arm; base; drug resistant virus; experience; genetic analysis; genetic approach; genome editing; improved; interest; macrophage; nerve stem cell; nervous system disorder; neurobehavioral disorder; neurocognitive disorder; novel; novel strategies; nuclease; preclinical study; promoter; prophylactic; public health relevance; resistant strain; screening; tool; vaccine development; viral DNA

DESCRIPTION (provided by applicant): According to the CDC, greater than 1.1 million people in the United States and more than 35 million people worldwide are infected with HIV-1. While the introduction of combined antiretroviral therapy, cART, has greatly improved survival rates among AIDS patients, a substantial portion of HIV-1 infected individuals remain at risk for the development of full blown AIDS as a result of reactivation of latently infected cells, partly due t nonadherence to medication and emergence of drug resistant viruses. Moreover, HIV-1 positive long term survivors continue to develop comorbidities including an accelerated aging process, neurocognitive disorders, heart failure, and others. From the virological point of view, as none of the current treatments suppress viral gene transcription, it is suspected that low, yet continuous, levels of viral early proteins with regulatory and pathogenic activities may contribute to the development of these quality of life threating illnesses. Sadly, none of the efforts toward the development of vaccines against HIV-1 have shown promising outcomes. Thus, curing of AIDS by eradicating the HIV-1 genome in infected subjects requires a novel strategy that is specific, highly effective, sustained, and irreversible. Recently, we have adapted a genetic approach using the clustered regulatory interspaced short palindromic repeat-assisted system (Cas) and a short complementary single-stranded RNA, called guide RNA or gRNA, which specifically targets the U3 region of the HIV-1 LTR promoter and precisely excises a segment of the viral regulatory sequence required for its expression. In addition, the employment of single and multiplex gRNA in our Cas system show promising results that include eradication of the entire HIV-1 genome in latently infected microglial cells, thus abrogating viral gene expression and transcription. Based on this preliminary observation, we propose to develop an RNA-guided Cas9 that acts as molecular scissors and, by disrupting various regions of the LTR and/or removing the entire viral genome, abrogates reactivation of the virus in macrophages, microglia and astrocytes which serve as the viral reservoir in the brain. Furthermore, we will explore the feasibility of our single and multiplex Cas9 system for use as a prophylactic compound in in vitro HIV-1 infection culture models. The outcome of this molecular genetic and virological approach will provide a solid platform for developing preclinical and clinical studies toward the treatment f AIDS and its associated neurological and neurobehavioral disorders.

描述（由申请人提供）：根据美国疾病控制与预防中心的数据，美国有超过110万人感染了HIV-1，全世界有超过3500万人感染了HIV-1。虽然采用抗逆转录病毒联合疗法（cART）大大提高了艾滋病患者的存活率，但由于潜伏感染的细胞重新激活，部分原因是不坚持服药和出现耐药病毒，很大一部分艾滋病毒-1感染者仍然面临发展为全面艾滋病的风险。此外，HIV-1阳性的长期幸存者继续出现合并症，包括加速衰老过程、神经认知障碍、心力衰竭等。从病毒学的角度来看，没有