Real-Time Spliced-RNA Detection to Quantify Latent HIV-Infected Cells in HAART Patients

实时剪接 RNA 检测可量化 HAART 患者中潜伏的 HIV 感染细胞

• 批准号: 9409647
• 负责人: Janet L Huie
• 金额: $ 100万
• 依托单位: JAN BIOTECH, INC.
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-03-01 至 2020-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9409647
• 关键词: 13 year old; AIDS clinical trial group; Acquired Immunodeficiency Syndrome; Adverse effects; Africa; Aftercare; Anti-Retroviral Agents; Biological Assay; Biotechnology; Blood; Blood specimen; CD4 Positive T Lymphocytes; Cause of Death; Cell Count; Cell Line; Cells; Centers for Disease Control and Prevention (U.S.); Characteristics; Clinical; Clinical Trials; Collaborations; Combined Modality Therapy; Consult; DNA; Detection; Development; Diagnostic; Disease remission; Enzymes; Evaluation; Genome; HIV; HIV Infections; HIV Seropositivity; HIV-1; HIV-associated neurocognitive disorder; Highly Active Antiretroviral Therapy; Human; Individual; Interruption; Letters; Measurement; Measures; Medicine; Methods; Monitor; Participant; Patients; Performance; Pharmacotherapy; Phase; Preclinical Testing; Predictive Analytics; Preparation; Procedures; Process; Public Health; RNA; RNA Splicing; RNA purification; Reaction; Research; Research Institute; Resources; Reverse Transcriptase Polymerase Chain Reaction; Ribonucleases; Risk; Sampling; Small Business Innovation Research Grant; Technology; Testing; Time; Universities; Validation; Viral; Viral Load result; Virion; Virus; Vulnerable Populations; assay development; base; commercialization; high throughput screening; improved; in vivo; interest; medical schools; molecular diagnostics; peripheral blood; pre-clinical; pre-exposure prophylaxis; preclinical evaluation; prevent; professor; sample collection; transmission process; treatment adherence; viral RNA; viral rebound

PROJECT SUMMARY ABSTRACT Public Health Problem In the U.S., 1,144,500 people aged 13 years and older are living with HIV infection, with approximately 180,900 (15.8%) others infected but undiagnosed. For most HIV-positive individuals, available treatments can only control HIV infection and delay progression to AIDS. HAART has substantial side effects and fails to prevent at least 50% of patients from developing HIV-1 associated neurocognitive disorders. Pre-exposure prophylaxis (PrEP) has been an exciting development for reduction of HIV acquisition risk and transmission, but it remains unclear when it is safe to discontinue PrEP treatment. The ultimate value of the SBIR Phase II project will demonstrate the ability of our technology to predict time to viral load rebound after treatment interruption, which remains a critical unmet need in the HIV cure field. Issues with Current Solutions & How Product Meets Unmet Needs Current methods of quantifying the latent reserve include the quantitative viral outgrowth assay (Q-VOA), PCR and RT-PCR. Q-VOA is a time and resource intensive procedure while RT-PCR can be used to detect viral RNA to 20-50 virus particles per mL and thus reduces the time to result of QVOA and is generally applicable for measuring viral load, but does not directly detect replication-competent latent HIV-infected cells. Jan Biotech’s HIV qLDR offers clear advantages including: 1) detection of latent reservoir cells in the blood without activation; 2) direct detection of RNA to improve quantitative ability that is lost in other assays; 3) nonenzymatic amplification, which relieves the need for RNA purification and its attendant loss of RNA; 4) detection of short segments of RNA, providing effective detection even in the presence of RNase degradative enzymes; 5) Jan Biotech’s qLDR PNA probes enable the use of short sequences complementary to highly conserved regions of the HIV-1 genome, which allows for breadth of detection across the highly genetically diverse HIV-1 subtypes. Summary of Approach The Phase I qLDR probe sets differentiated between latent and activated HIV-infected cells in peripheral blood, which has great promise for accurate measurement of the level of the replication-competent latent reservoir. The Phase II Specific Aims will demonstrate the ability of the assay to predict time to viral load rebound after treatment interruption using ex vivo and in vivo samples. Testing will evaluate the utility of the assay as a clinical endpoint to guide treatment interruption decisions and facilitate HIV remission and cure discovery. Collaborators and Unique Resources Jan Biotech, Inc., with expertise in molecular diagnostic development, will consult with Jonathan Li, MD, MMSc Assistant Professor of Medicine, Harvard Medical School, in a AIDS Clinical Trials Group (ACTG) study. Judith Currier, MD, MSc, Vice Chair of the ACTG Network will oversee sample collection from the treatment interruption study, A5345; Q-VOA cross-validation testing will be performed by Southern Research Institute. Phase II Specific Aims Specific Aim 1: HIV qLDR prediction of replication-competent latent reservoir Task 1: Optimization of qLDR probe sets Task 2: Ex vivo rebound after treatment interruption (TI) Specific Aim 2: HIV qLDR prediction of HIV rebound during Analytical Treatment Interruption Task 1: HIV qLDR prediction of Viral Rebound after Analytical Treatment Interruption (ATI) Task 2: Comparison of HIV qLDR to other assays used in ATI study Specific Aim 3: Preclinical Validation of HIV qLDR Task 1: Validation of qLDR performance characteristics Task 2: QVOA validation Market after Phase II Completion Both Gilead and Bionor are commercially interested in the technology and look forward to results generated from the Phase II and the ACTG study. Regulatory approval will be sought for commercialization.

项目摘要摘要 公共卫生问题 在美国，1,144,500名13岁及以上的人患有艾滋病毒感染，约有180,900人 （15.8％）其他被感染但未诊断的人。对于大多数艾滋病毒阳性的人，可用的治疗只能 控制艾滋病毒感染并延迟进展为艾滋病。 Haart具有重大副作用，无法预防 患有HIV-1相关的神经认知疾病的患者中，至少有50％的患者。暴露前预防 （PREP）是减少艾滋病毒获取风险和传播的令人兴奋的发展，但仍然是 尚不清楚何时安全停止准备治疗。 SBIR II期项目的最终价值将证明我们技术预测时间的能力 治疗中断后的病毒载荷反弹，这在HIV治疗领域仍然是至关重要的未满足需求。 当前解决方案的问题以及产品如何满足未满足的需求 量化潜在储备的当前方法包括定量病毒生长测定（Q-VOA），PCR 和RT-PCR。 Q-VOA是一个时间和资源密集型程序，而RT-PCR可用于检测病毒 RNA至20-50个病毒颗粒，因此减少了QVOA的时间，通常适用 用于测量病毒载量，但不能直接检测到具有复制能力的潜在艾滋病毒感染细胞。扬 生物技术的HIV QLDR提供明显的优势，包括：1）检测没有血液中的潜在储层细胞 激活; 2）直接检测RNA以提高其他测定中丢失的定量能力； 3）非酶 放大，可缓解RNA纯化的需求及其随之而来的RNA损失； 4）检测短 RNA的片段，即使存在RNase降解酶，也提供有效的检测； 5）1月 Biotech的QLDR PNA问题使得可以将短序列完善使用到高度组成的区域 HIV-1基因组，可以在高度遗传多样的HIV-1亚型中进行广泛的检测。 方法摘要 I阶段QLDR探针设置在周围血液中的潜在和活化的HIV感染细胞区分， 这具有准确测量复制能力的潜在储层水平的巨大希望。 第二阶段特定的目的将证明测定法预测后的病毒载荷时间的能力 使用离体和体内样品的处理中断。测试将评估测定的效用 指导治疗中断决策并促进HIV缓解和治愈发现的临床终点。 合作者和独特的资源 Jan Biotech，Inc。具有分子诊断开发方面的专业知识，将咨询Jonathan Li，MD，MMSC 哈佛医学院医学助理教授，在AIDS临床试验小组（ACTG）研究中。朱迪思 ACTG网络副主席Currier，医学博士，MSC，将监督处理中的样本收集 中断研究，A5345；南方研究所将进行Q-VOA交叉验证测试。 第二阶段特定目标 特定目的1：HIV QLDR复制能力潜在储层的预测 任务1：QLDR探针集的优化 任务2：治疗中断后的体内反弹（TI） 特定目标2：分析治疗过程中HIV反弹的HIV QLDR预测 任务1：分析治疗中断后病毒反弹的HIV QLDR预测（ATI） 任务2：HIV QLDR与ATI研究中使用的其他测定法的比较 特定目的3：HIV QLDR的临床前验证 任务1：验证QLDR性能特征 任务2：QVOA验证 第二阶段完成后市场 Gilead和Bionor都对技术感兴趣，并期待产生的结果 来自第二阶段和ACTG研究。将感觉到商业化的监管批准。