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

基本信息

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
项目类别：
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

项目摘要

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.

项目概要摘要公共卫生问题在美国，1,144,500 名 13 岁及以上人群感染艾滋病毒，其中约 180,900 人感染艾滋病毒 (15.8%) 其他已感染但未确诊的人对于大多数艾滋病毒阳性者来说，现有的治疗只能是有效的。控制艾滋病毒感染并延缓艾滋病毒的进展有很大的副作用，并且无法预防。至少 50% 的患者患有 HIV-1 相关的神经认知障碍。 PrEP（暴露前预防）对于降低艾滋病毒感染风险和传播来说是一项令人兴奋的进展，但它仍然存在尚不清楚何时停止 PrEP 治疗是安全的。 SBIR 二期项目的最终价值将证明我们的技术有能力预测实现目标的时间治疗中断后病毒载量反弹，这仍然是艾滋病毒治疗领域未满足的一个关键需求。当前解决方案的问题以及产品如何满足未满足的需求目前量化潜在储备的方法包括定量病毒生长 (Q-VOA)、PCR 检测 RT-PCR 是一种时间和资源密集型程序，而 RT-PCR 可用于检测病毒。 RNA 为每毫升 20-50 个病毒颗粒，从而缩短获得 QVOA 结果的时间，普遍适用用于测量病毒载量，但不能直接检测具有复制能力的潜伏 HIV 感染细胞。 Biotech 的 HIV qLDR 具有明显的优势，包括：1）无需检测即可检测血液中的潜伏储存细胞激活；2) 直接检测 RNA，以提高其他测定中失去的定量能力；3) 扩增，从而减少了 RNA 纯化的需要以及随之而来的 RNA 损失 4) 短片段的检测； RNA 片段，即使在 RNase 降解酶存在的情况下也能提供有效的检测；5) Jan Biotech 的 qLDR PNA 探针能够使用与高度保守的区域互补的短序列 HIV-1 基因组，可广泛检测遗传多样性的 HIV-1 亚型。方法总结 I 期 qLDR 探针可区分外周血中潜伏和激活的 HIV 感染细胞，这对于精确测量具有复制能力的潜在储存库的水平具有很大的前景。第二阶段的具体目标将证明该检测方法能够预测病毒载量反弹后的时间使用离体和体内样品进行治疗中断评估该测定法的实用性。治疗的临床终点指导中断决策并促进艾滋病毒缓解和治愈方法的发现。合作者和独特资源 Jan Biotech, Inc. 拥有分子诊断开发方面的专业知识，将咨询医学博士、理学硕士 Jonathan Li 哈佛医学院医学助理教授朱迪思 (Judith) 参与艾滋病临床试验组 (ACTG) 研究。 ACTG 网络副主席、医学博士、理学硕士 Currier 将监督治疗的样本收集中断研究，A5345；Q-VOA交叉验证测试将由南方研究院进行。第二阶段具体目标具体目标 1：具有复制能力的潜在病毒库的 HIV qLDR 预测任务 1：qLDR 探针组的优化任务 2：治疗中断 (TI) 后的离体反弹具体目标 2：分析治疗中断期间 HIV 反弹的 HIV qLDR 预测任务 1：分析治疗中断 (ATI) 后病毒反弹的 HIV qLDR 预测任务 2：HIV qLDR 与 ATI 研究中使用的其他检测方法的比较具体目标 3：HIV qLDR 的临床前验证任务 1：验证 qLDR 性能特征任务 2：QVOA 验证二期竣工后的市场吉利德和 Bionor 都对该技术产生商业兴趣，并期待产生的结果将寻求第二阶段和 ACTG 研究的监管批准以进行商业化。