PANDAA for rapid genotyping of HIV-1 infected patients failing protease inhibitor ART in resource-limited settings

PANDAA 用于在资源有限的环境中对未能接受蛋白酶抑制剂 ART 治疗的 HIV-1 感染患者进行快速基因分型

• 批准号: 9256022
• 负责人: Iain James MacLeod
• 金额: $ 99.8万
• 依托单位: ALDATU BIOSCIENCES, INC.
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-01-17 至 2019-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9256022
• 关键词: Adherence; Adult; Anti-Retroviral Agents; Biological Assay; Biological Sciences; Child; Clinical; Communicable Diseases; Cost Savings; Country; Data; Decision Making; Detection; Development; Diagnostic; Dideoxy Chain Termination DNA Sequencing; Drug Costs; Drug resistance; Drug-sensitive; Ensure; Exhibits; Failure; Feasibility Studies; Formulation; Freeze Drying; Future; Genetic Polymorphism; Genomics; Genotype; Guidelines; HIV; HIV drug resistance; HIV resistance; HIV-1; In Vitro; Institutes; Laboratories; Measures; Molecular Cloning; Mutation; Patient Care; Patients; Performance; Pharmaceutical Preparations; Phase; Point Mutation; Positioning Attribute; Preparation; Production; Protease Inhibitor; Quality Control; RNA; Reagent; Regimen; Reproducibility; Resistance; Resources; Sampling; Sensitivity and Specificity; Site; Small Business Innovation Research Grant; Specificity; System; Target Populations; Technology; Testing; Therapeutic; Time; Treatment Failure; Validation; Variant; Vertebral column; Viral; Viral Load result; base; blind; clinical decision-making; cost; cost effective; design; drug testing; improved; innovation; low and middle-income countries; method development; non-nucleoside reverse transcriptase inhibitors; novel; patient stratification; prevent; programs; prototype; research and development; resistance mutation; thermostability; virology

The 2015 WHO guidelines recommend anti-retroviral therapy (ART) for all HIV-infected people, and massive efforts to expand ART access will result in >30M adults and >1.7M children on ART by 2020, at a cost of almost US$53B. Consequently, the number of patients prescribed alternative, protease-inhibitor (PI)-based ART following failure of first-line regimens will also increase. Studies of patients on PI-based ART in resource-limited countries have shown high rates of virologic failure at 12 and 24 months on treatment, and patients maintained on failing PI-based regimens accumulate drug resistance mutations (DRMs) that hamper current and future treatment options. Resource constraints often limit the ability to collect genotype information, and genotype-blind drug switching has been shown to be cost-ineffective when compared to stratifying patients based on resistance genotype. Thus, there is an urgent unmet need for an affordable HIV genotyping option for patients failing PI- based ART. Focused genotyping could significantly enhance patient care by: 1) optimizing the NRTI backbone in patients failing a PI-based regimen with only NRTI resistance; or 2) preventing ART switches in patients failing without resistance until adherence issues are rectified. Aldatu’s Pan-Degenerate Amplification and Adaptation (PANDAA™) technology is a novel point mutation assay that enables inexpensive and high-throughput focused genotypic resistance testing, and such an approach could be cost-saving for national ART programs. PANDAA compensates for high intra- and inter-patient HIV genomic variability by removing secondary polymorphisms, minimizing their impact on qPCR sensitivity/specificity, and enabling qPCR for HIV genotyping for the first time. Feasibility studies have demonstrated that PANDAA: 1) detects NNRTI and NRTI-resistant HIV variants with >99% sensitivity; 2) is HIV subtype-independent; and 3) can be multiplexed to simultaneously quantify resistance at multiple genomic positions. Aldatu has pioneered the commercial development of PANDAA and established a reagent formulation that allows for the production of PANDAA-based diagnostics in a thermostable, sample- ready format. In this Phase II project, Aldatu will apply the PANDAA technology to the development of PANDAA PIDR+, a rapid, low-cost, thermostable test for detection of drug resistance in patients failing a PI-based ART regimen, which can radically improve clinical decision-making in low- and middle-income countries. Through the aims proposed here, we will 1) experimentally validate the design of PANDAA reagents to quantify mutations associated with protease inhibitor resistance comprising 10% of the viral quasispecies; 2) establish an extensive, collaborative proficiency panel of drug resistant and drug sensitive HIV-1 isolates for PANDAA PIDR+ validation; 3) assess PANDAA PIDR+ using established performance criteria for HIV drug resistance genotyping and produce PANDAA PIDR+ under GMP conditions; and 4) verify that end-user, multi-site implementation of PANDAA PIDR+ is highly reproducible. The first of its kind, a validated, GMP-produced PANDAA PIDR+ test kit from Aldatu will be poised to capture a significant share of this rapidly growing diagnostic market opportunity.

2015年指南建议所有感染HIV的人的抗逆转录病毒疗法（ART），并大量 扩大艺术访问的努力将导致> 30m的成年人和> 170万儿童在2020年的艺术中，几乎为 US $ 53B。因此，处方替代蛋白酶抑制剂（PI）的ART的患者数量 一线方案失败后，也将增加。对资源有限的基于PI的ART的患者研究 国家在治疗12和24个月的病毒学衰竭率很高，患者保持 基于PI失败的方案会累积阻碍电流和未来的耐药性突变（DRM） 治疗选择。资源限制通常会限制收集基因型信息的能力和基因型盲 与基于阻力进行分层的患者相比，药物转换已被证明具有成本效益 基因型。这是针对PI-失败的患者的负担得起的HIV基因分型选项的紧急需求 基于艺术。聚焦基因分型可以通过以下方式显着增强患者护理：1）优化NRTI主链 在失败的基于PI的治疗方案的患者中，仅NRTI耐药性；或2）防止患者失败的艺术切换 在纠正依从性之前，没有抵抗。 Aldatu的泛泛化和适应 （PANDAA™）技术是一种新颖的点突变测定法，可实现廉价且高通量的重点 基因型抗性测试以及这种方法可能会节省国家艺术计划。熊达 通过去除次级多态性，可以补偿高较高的患者内和患者间的HIV基因组变异性， 最小化其对QPCR敏感性/特异性的影响，并首次启用QPCR进行HIV基因分型。 可行性研究表明，熊猫：1）检测NNRTI和NRTi耐药的HIV变体 > 99％的灵敏度； 2）是HIV亚型独立的； 3）可以多路复用以同时量化电阻 在多个基因组位置。 Aldatu开创了Pandaa的商业发展，并建立了 一种试剂公式，允许在可热稳定的样品中生产基于熊猫的诊断。 准备格式。在此第二阶段项目中，Aldatu将把熊猫技术应用于熊达的开发 PIDR+，一种快速，低成本的可热稳定测试，用于检测失败的基于PI ART的患者的耐药性 方案，可以从根本上改善低收入和中等收入国家的临床决策。通过 在这里提出的目的，我们将1）实验验证熊猫试剂的设计以量化突变 与蛋白酶抑制剂耐药性完成10％的病毒式准蛋白酶有关； 2）建立广泛的 抗药性和药物敏感的HIV-1分离株的协作能力小组进行熊猫PIDR+验证； 3）使用确定的HIV耐药性基因分型和 在GMP条件下产生pandaa pidr+； 4）验证最终用户的多站点实现 Pandaa Pidr+高度可重现。同类产品中的第一个是经过验证的GMP生产的Pandaa Pidr+测试套件 Aldatu将被毒化，以捕获这一快速增长的诊断市场机会中的很大一部分。