Novel Assays for Measuring Susceptibility to Inhibitors that Target HIV1 Virion A

用于测量针对 HIV1 病毒粒子 A 的抑制剂敏感性的新方法

• 批准号: 7688713
• 负责人: CHRISTOS J PETROPOULOS
• 金额: $ 29.96万
• 依托单位: MONOGRAM BIOSCIENCES, INC.
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-03-01 至 2011-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7688713
• 关键词: Amendment; Amino Acid Sequence; Anti-Retroviral Agents; Base Sequence; Biological Assay; Capsid; Cells; Chemistry; Clinic; Clinical; Clinical Research; Clinical Trials; Code; Codon Nucleotides; Collection; DNA; DNA Restriction Enzymes; DNA Sequence; Data; Data Reporting; Development; Digestion; Drug resistance; Engineering; Environment; Epitopes; Evaluation; Funding; Gagging; Generations; Genes; Genetic; Genomics; Goals; Guidelines; HIV; HIV drug resistance; HIV vaccine; HIV-1; HIV-1 Reverse Transcriptase; HIV-1 protease; Immune; Immune response; Infection; Integrase; Integrase Inhibitors; Laboratories; Ligation; Luciferases; Measurable; Measures; Minor; Molecular Cloning; Monitor; Mutation; New Agents; Nucleic acid sequencing; Nucleocapsid; Nucleotides; Oligonucleotides; Parents; Patient Selection; Patients; Peptide Hydrolases; Performance; Pharmaceutical Preparations; Phase; Phase I Clinical Trials; Phase II Clinical Trials; Plasma; Plasmids; Population; Predisposition; Preparation; Procedures; Process; Protease Inhibitor; Proteins; Quality Control; RNA-Directed DNA Polymerase; Reaction; Recombinants; Reporter; Reporter Genes; Reporting; Reproducibility; Research; Resistance; Reverse Transcriptase Polymerase Chain Reaction; SP1 gene; Sampling; Sensitivity and Specificity; Sequence Analysis; Site; Site-Directed Mutagenesis; Software Tools; Source; Specific qualifier value; Specificity; Staging; System; T-20; Technology; Testing; Transfection; Treatment Protocols; Vaccine Design; Validation; Variant; Viral; Viral Load result; Viral Vector; Virion; Virus; Virus Assembly; Virus Replication; base; clinical practice; commercialization; cost; design; disorder subtype; drug candidate; env Genes; experience; fitness; gag Gene Products; high throughput screening; improved; inhibitor/antagonist; mutant; novel; pre-clinical; prospective; public health relevance; recombinant virus; research study; resistance mutation; restriction enzyme; tool; validation studies; vector

DESCRIPTION (provided by applicant): The overall goal of this project is to develop novel, robust, high-throughput assays to evaluate the susceptibility of to inhibitors of HIV-1 Gag and protease proteins, with particular emphasis on virion assembly and maturation. These assays will be based on proven technologies underlying Monogram Bioscience's widely used PhenoSense HIVTM and GeneSeq HIVTM drug resistance tests, i.e. recombinant virus-based and nucleic acid sequencing-based assays first developed to evaluate inhibitors of HIV-1 protease and reverse transcriptase. Currently, several assembly/maturation inhibitors are undergoing evaluation in pre-clinical and clinical studies, e.g. bevirimat (Panacos), vivecon (Myriad Genetics). Consequently, reliable, high-throughput assays are needed to support the development efforts of this promising new class of antiretroviral drugs, and eventually, their use in routine clinical practice. Since their implementation in the Monogram Clinical Reference Laboratory and commercialization in 2000, the PhenoSense HIV and GeneSeq HIV assays have been used to characterize in excess of 150,000 patient samples. In addition, both assays have been used extensively to support phase III evaluations of antiretroviral drug candidates over the past eight years, both to characterize resistance to new agents and to guide the selection of optimized background regimens. In 2003, Monogram successfully adapted the PhenoSense and GeneSeq technologies to evaluate inhibitors of HIV-1 entry (e.g. enfuvirtide, maraviroc, vivcriviroc, ibalizumab, PRO-140), and this year we completed validations for two new assays to monitor resistance to integrase inhibitors (e.g. raltegravir, elvitegravir). This application describes a straightforward, tried and true strategy to develop PhenoSense and GeneSeq HIV drug resistance assay systems that are capable of evaluating inhibitors of HIV-1 Gag and protease proteins; specifically inhibitors that target virus assembly and maturation. Because these new assay systems will include the immuno- dominant Gag region, they arelikely to prove valuable to the study of cellular immune responses. Such studies may prove valuable to vaccine design and evaluation. PUBLIC HEALTH RELEVANCE: Successful completion of the funding proposal will result in the development and validation of new genotypic and phenotypic assays that will enable the characterization of viruses that are resistant to HIV-1 assembly and maturation inhibitors and will help guide antiretroviral treatment decisions for HIV-1 infected patients. The assay will also greatly advance the genotypic and phenotypic characterization of gag-specific CTL escape variants and will prove valuable to the design and development of improved HIV vaccines and immune-modulating therapies.

描述(由申请人提供)：该项目的总体目标是开发新的、强大的、高通量的检测方法，以评估HIV-1 Gag和蛋白酶蛋白抑制剂的敏感性，特别强调病毒粒子的组装和成熟。这些检测将基于Mongraph Bioscience广泛使用的PhenoSense HIVTM和GeneSeq HIVTM耐药性测试所依据的成熟技术，即最初为评估HIV-1蛋白酶和逆转录酶抑制剂而开发的基于重组病毒和核酸测序的分析方法。目前，几种组装/成熟抑制剂正在进行临床前和临床研究中的评估，如bevirimat(Panacos)，viveon(Myriad Genetics)。因此，需要可靠、高通量的分析来支持这类有前景的新型抗逆转录病毒药物的开发工作，并最终将其用于常规临床实践。自从在Mongraph临床参考实验室实施并于2000年商业化以来，PhenoSense HIV和GeneSeq HIV检测已经被用于鉴定超过15万个患者样本。此外，在过去八年中，这两种检测方法被广泛用于支持抗逆转录病毒候选药物的第三阶段评估，既用于表征对新药物的耐药性，也用于指导优化背景方案的选择。2003年，Mongraph成功地采用了PhenoSense和GeneSeq技术来评估HIV-1进入的抑制剂(例如恩福韦肽、马拉韦罗、vivcriviroc、ibalizumab、PRO-140)，今年我们完成了两种新的检测方法的验证，以监测对整合酶抑制剂(例如raltegravir、elvitegravir)的耐药性。本申请描述了一种简单、可靠的策略来开发PhenoSense和GeneSeq HIV耐药性测试系统，该系统能够评估HIV-1 Gag和蛋白酶蛋白的抑制剂；特别是针对病毒组装和成熟的抑制剂。由于这些新的检测系统将包括免疫主导的Gag区域，它们很可能被证明对细胞免疫反应的研究有价值。这些研究可能被证明对疫苗设计和评估有价值。与公共卫生相关：供资提案的成功完成将导致新的基因和表型分析的开发和验证，这些分析将能够表征对艾滋病毒-1组装和成熟抑制剂具有抗药性的病毒，并将有助于指导艾滋病毒-1感染者的抗逆转录病毒治疗决定。该分析还将极大地提高Gag特异性CTL逃逸变异体的基因型别和表型特征，并将被证明对改进的HIV疫苗和免疫调节疗法的设计和开发有价值。