Standardizable Whole Blood Culture for Evaluating HIV Eradication Strategies

用于评估 HIV 根除策略的标准化全血培养

• 批准号: 9139798
• 负责人: Heidi A Kay
• 金额: $ 41.64万
• 依托单位: JERICHO SCIENCES, LLC
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-08-10 至 2018-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9139798
• 关键词: AIDS/HIV problem; Acquired Immunodeficiency Syndrome; Anti-Retroviral Agents; Antiviral Agents; Benefits and Risks; Biological Assay; Biological Markers; Blood; Blood Cells; Blood Volume; Blood donor; Blood specimen; Budgets; Cell Therapy; Cells; Chronic Disease; Clinic; Clinical; Clinical Research Associate; Clinical Treatment; Clinical Trials; Collection; Data; Detection; Development; Devices; Diagnostic; Disease; Disease remission; Drug Design; Drug resistance; Ensure; Evaluation; Genetic Transcription; Genotype; Guidelines; HIV; HIV Infections; HIV-1; Head; Hour; Human immunodeficiency virus test; Immune; Individual; Infection; Inflammatory; Intervention; Knowledge; Lead; Legal patent; Letters; Measures; Mediating; Mission; Modeling; National Institute of Allergy and Infectious Disease; Pathway interactions; Patient Monitoring; Patient risk; Patients; Pharmaceutical Preparations; Phase; Preclinical Drug Evaluation; Production; Proliferation Marker; Protocols documentation; Proviruses; Reproducibility; Research; Sampling; Science; Sensitivity and Specificity; Small Business Innovation Research Grant; Source; System; T-Lymphocyte; Technology; Testing; Therapeutic; Therapeutic Effect; Therapeutic Intervention; Therapeutic Uses; Translating; Tube; Viral; Viral reservoir; Virion; Virus; Virus Activation; Whole Blood; Work; clinically relevant; curative treatments; cytokine; design; drug development; effective therapy; experience; immunological status; immunoregulation; individual patient; inhibitor/antagonist; meetings; next generation; novel diagnostics; peripheral blood; pre-clinical; programs; prospective; public health relevance; purge; research clinical testing; response; sample collection; skills; success; therapeutic target; therapy development; tool

 DESCRIPTION (provided by applicant): The impetus for new "curative" approaches for HIV-AIDS lacks adequate clinical tools to monitor patient responses to therapeutics targeting the persistent viral reservoir. We therefore propose to develop a standardizable, integrated, clinically-relevant whole blood culture assay to evaluate HIV eradication therapeutics ex vivo by directly measuring responses of primary peripheral blood cells under internally- referenced patient-specific matched control and therapeutic conditions. The proposed system will quantify single-cycle virus production from infected primary blood cells comprising the entire peripheral blood immunological compartment. The relative difference in virus production under latency-activating conditions versus unstimulated matched cultures represents the target replication-competent provirus residing in latently- infected primary blood cells. The culture assay will support ex vivo testing of latency-activating, cytolytic, immunomodulatory and other "curative" therapeutic approaches required by the FDA for therapeutic success. This standardizable, readily-accessible whole blood culture assay for testing HIV eradication strategies will provide an enabling, next-generation research, clinic and patient-management platform. The Jericho team proposes to validate the feasibility of developing such a platform here via this ambitious Phase I SBIR project (expanded scope and budget). The envisioned integrated blood-collection system for HIV-infected whole blood culture is a closed device requiring less than 10.0 mL of donor blood per collection tube, directly obtained by a phlebotomist using a standard blood draw. After up to 72 hours of incubation at 37⁰C in a proprietary media, viable cells, soluble factors and virions can be co-collected and measured from these primary blood cultures to provide correlative biomarkers. The proposed product will allow for comparison of different test conditions (e.g., latency activators) and therapeutics (e.g., suppressive drugs, cytokines) across multiple internally-referenced culture tubes, specific to the infection status and immunological response of the individual. We propose to demonstrate the feasibility of standardizing and developing this platform to evaluate endpoints for HIV eradication strategies by characterizing virus production, cellular markers, and soluble factors from 45 HIV-infected donor whole blood sample collections. We will first quantify and characterize reproducibility of our stimulated virus production culture system within a coefficient of variance ≤14% between replicate, matched collection tubes. We will then demonstrate the potential utility of the characterized platform using two selected compounds known to stimulate latently-infected cells. Preclinical therapeutic optimization translates directly to clinical monitoring and patient management, having identified relevant conditions and associated biomarkers using the proposed whole blood culture assay platform. As HIV is now moving from a manageable chronic disease to a "cure," novel diagnostics will be increasingly required to ensure that the best treatments are used to optimize efficacy and to reduce patient risks.

 描述（由申请人提供）：HIV-AIDS新“治愈”方法的动力缺乏足够的临床工具来监测患者对靶向持久性病毒储库的治疗的反应。因此，我们建议开发一种可重复使用的、集成的、临床相关的全血培养测定法，以通过在内部参考的患者特异性匹配对照和治疗条件下直接测量原代外周血细胞的反应来离体评价HIV根除治疗剂。所提出的系统将量化单循环病毒生产从感染的原代血细胞，包括整个外周血免疫室。潜伏激活条件下的病毒产生与未刺激的匹配培养物相比的相对差异代表了存在于潜伏感染的原代血细胞中的靶复制能力前病毒。培养试验将支持FDA要求的潜伏激活、细胞溶解、免疫调节和其他“治愈性”治疗方法的离体测试，以获得治疗成功。这种用于测试HIV根除策略的可扩展、易于获得的全血培养检测方法将提供一个支持的下一代研究、临床和患者管理平台。Jericho团队建议通过这个雄心勃勃的第一阶段SBIR项目（扩大范围和预算）来验证开发这样一个平台的可行性。设想的用于HIV感染的全血培养的集成血液采集系统是一种封闭装置，每个采集管需要少于10.0 mL的供体血液，由采血师使用标准抽血直接获得。在37 ° C下在专有培养基中孵育长达72小时后，可以从这些原代血液培养物中共同收集和测量活细胞、可溶性因子和病毒体，以提供相关的生物标志物。申报产品将允许比较不同的测试条件（例如，潜伏激活剂）和治疗剂（例如，抑制药物、细胞因子），特异性针对个体的感染状态和免疫应答。我们建议证明标准化和开发该平台的可行性，以评估艾滋病毒根除策略的终点，通过表征45个艾滋病毒感染的供体全血样本采集的病毒产生，细胞标志物和可溶性因子。我们将首先量化和表征我们的刺激病毒的再现性， 生产培养系统的重复匹配采集管之间的变异系数≤14%。然后，我们将使用已知刺激潜伏感染细胞的两种选定化合物来证明表征平台的潜在效用。临床前治疗优化直接转化为临床监测和患者管理，使用拟议的全血培养测定平台鉴定相关病症和相关生物标志物。随着艾滋病毒正在从一种可控制的慢性疾病转变为一种“治愈”疾病，将越来越需要新的诊断方法，以确保使用最佳治疗方法来优化疗效并降低患者风险。