Pediatric Latent HIV Reservoir Characterization and Quantification Assay

儿科潜伏 HIV 储库特征和定量分析

• 批准号: 10761022
• 负责人: Janet L Huie
• 金额: $ 30万
• 依托单位: JAN BIOTECH, INC.
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10761022
• 关键词: AIDS clinical trial group; Acute; Adult; Aftercare; Age; Antibodies; Base Sequence; Binding; Biological Assay; Biological Markers; Biotechnology; Birth; Blood; Blood Volume; Blood specimen; CD4 Positive T Lymphocytes; Calibration; Cell Count; Cells; Characteristics; Chemicals; Child; Childhood; Clinical; Clinical Trials; Cohort Studies; Combined Modality Therapy; Complementary DNA; Consult; Detection; Development; Disease remission; Doctor of Philosophy; Evaluation; Fluorescence; Fluorescence-Activated Cell Sorting; HIV; HIV Infections; HIV-1; Human; Individual; International; International Maternal Pediatric Adolescent AIDS Clinical Trials; Interruption; Marketing; Measurement; Measures; Medicine; Methods; Modernization; Monitor; Oligonucleotides; Participant; Performance; Peripheral Blood Mononuclear Cell; Persons; Phase; Physicians; Population; Preparation; Public Health; RNA; Recombinants; Reproducibility; Resource-limited setting; Resources; Sampling; Small Business Innovation Research Grant; Specificity; Technology; Testing; Time; Treatment Efficacy; United States National Institutes of Health; Universities; Validation; amplification detection; antiretroviral therapy; clinically relevant; cohort; commercialization; cost; curative treatments; design; detection limit; improved; interest; internal control; latent HIV reservoir; medical schools; molecular diagnostics; multiplex assay; pediatric human immunodeficiency virus; professor; transmission process

PROJECT SUMMARY ABSTRACT Public Health Problem Modern antiretroviral therapy (ART) combinations greatly improve survival of HIV infected individuals and reduce HIV transmission. However, there are still 38 million people living with HIV (PWH) worldwide and of those 1.8 million are children under the age of 15, highlighting the imperative need for curative therapies. In limited resource areas, where pediatric HIV is most common, the use of new assays that require less blood volumes, lower cost and less labor-intensive methods are necessary to facilitate clinical trials aimed at reducing the size of the latent reservoir and/or to achieve a cure or ART-free remission. The ultimate value of the SBIR Phase I project will demonstrate the ability of our technology to characterize and quantify the pediatric HIV-1 reservoir, which remains a critical unmet need in the HIV cure field. Issues with Current Solutions & How Product Meets Unmet Needs RNAamp allows direct detection of RNA, which provides the most accurate measure of the HIV replication competent reservoir. The assay is proven to be faster and have better reliability and quantification than HIV reservoir assays based on QVOA, RT-qPCR, RT-ddPCR, iSCA and IPDA. RNA target amplification and detection is accomplished by oligonucleotide-templated photoreduction using short, sequence-specific profluorophore and catalytic probes. Reproducibility is outstanding (4% variance between replicates) and Limit of Detection (LoD) is 1 HIV-infected cells/1 x 106 PBMCs or 1-10 RNA copies/mL, using a standard blood draw (10-30 mL). Jan Biotech’s RNAamp quick, direct RNA test, with its use of small blood samples, ease of sample preparation, reliability and sensitivity, quantitative ability and short time to result, is ideally suited for characterization and quantification of the rebound-competent pediatric HIV reservoir. Summary of Approach Jan Biotech has developed an assay with a prediction accuracy of 0.92 for time to rebound after treatment interruption for early/acute-treated adult participants. The assay, RNAamp, utilizes nonezymatic, chemical amplification of cell-associated HIV RNA targets to provide quick, sensitive, and quantitative measurement of the HIV reservoir. No other assay, including QVOA, IPDA, iSCA, RT-qPCR, or RT-ddPCR, provided a prediction accuracy of greater than 0.2 for the same adult participants. This proposal is to develop RNAamp for cross- subtype (i.e., non-B subtype) HIV reservoir detection, specifically for pediatric testing, for which the use of small samples easily tested by RNAamp is crucial. Pediatric HIV-infected individuals are typically treated during early HIV infection, often treated at birth, and physician-monitored or spontaneous treatment interruption is more frequent in this population. Pediatric treatment efficacy and treatment interruption clinical trials typically include non-B subtype HIV-infected participants. 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 and Harris Gelbard, MD, PhD, University of Rochester Medical School. Phase I Specific Aims Specific Aim 1: Develop HIV-1 cross-subtype (CS) RNAamp multiplexed assay Specific Aim 2: Correlate CS-RNAamp to Pol RT-qPCR using non-B subtype HIV-infected cells Specific Aim 3: Pediatric human sample testing using CS-RNAamp HIV reservoir characterization Market after Phase II Completion Both Gilead and Merck are commercially interested in the technology. Regulatory approval will be sought for commercialization.

项目总结摘要 公共卫生问题 现代抗逆转录病毒疗法(ART)的组合极大地提高了艾滋病毒感染者的存活率，并减少了 艾滋病毒的传播。然而，全世界仍有3800万艾滋病毒(PWH)携带者，其中1.8人 100万是15岁以下的儿童，这突显了治疗方法的迫切需要。在有限的 在儿科艾滋病毒最常见的资源领域，使用需要较少血量的新检测方法， 更低的成本和更少的劳动密集型方法是必要的，以促进旨在减少规模的临床试验 和/或达到治愈或无抗逆转录病毒治疗的缓解。 SBIR第一阶段项目的最终价值将证明我们的技术能够表征和 量化儿童艾滋病毒-1宿主，这在艾滋病毒治疗领域仍然是一个关键的未得到满足的需求。 当前解决方案存在的问题以及产品如何满足未满足的需求 RNAAmp允许直接检测RNA，这提供了对HIV复制的最准确测量 称职的储集层。事实证明，该检测方法比HIV更快、更可靠、更定量 基于QVOA、RT-qPCR、RT-ddPCR、ISCA和IPDA的储层分析。RNA靶向扩增和 检测是通过使用短的、序列特异性的寡核苷酸模板光还原来完成的 前荧光团和催化探针。再现性突出(复制之间的差异为4%)和限制 使用标准血样，检测的灵敏度(LOD)为1个HIV感染细胞/1×106个PBMC或1-10个RNA拷贝/毫升 (10-30毫升)。 Jan Biotech的RNAamp快速、直接的RNA检测，具有使用少量血液样本、易于制备样本、 可靠性和敏感度、定量能力和结果时间短，非常适合表征和 量化具有反弹能力的儿科艾滋病毒储备库。 方法概述 Jan Biotech开发了一种检测方法，对治疗后反弹的时间的预测精度为0.92 对接受早期/急性治疗的成人参与者的中断。这种名为RNAamp的检测方法使用的是非酶、化学 扩增细胞相关的HIV RNA靶标以提供快速、灵敏和定量的测量 艾滋病病毒携带者。包括QVOA、IPDA、ISCA、RT-qPCR或RT-ddPCR在内的其他检测都不能提供预测 对于相同的成年参与者，准确率大于0.2。这项建议是为了开发用于交叉的RNAAmp 亚型(即非B亚型)HIV感染者的检测，专门用于儿科检测，对于其中的使用较少 容易被RNAamp检测的样本是至关重要的。儿童艾滋病毒感染者通常在早期接受治疗 艾滋病毒感染，通常在出生时接受治疗，而医生监测或自发中断治疗的情况更多 在这个人群中经常发生。儿科治疗疗效和治疗中断临床试验通常包括 非B亚型艾滋病毒感染者。 合作者和独特资源 Jan Biotech，Inc.在分子诊断开发方面拥有专业知识，将与Jonathan Li，MD，MMSc进行咨询 哈佛医学院医学助理教授，参与艾滋病临床试验小组(ACTG)的研究 哈里斯·格尔巴德，医学博士，罗切斯特大学医学院博士。 第一阶段的具体目标 具体目标1：建立HIV-1交叉亚型(CS)RNAAmp多重检测方法 特定目的2：利用非B亚型HIV感染细胞将CS-RNAAmp与Pol RT-qPCR相关联 具体目标3：使用CS-RNAamp艾滋病毒储备库特征进行儿科人体样本测试 第二期完成后的市场 吉利德和默克在商业上都对这项技术感兴趣。将寻求监管部门的批准 商业化。