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年世卫组织指南建议对所有艾滋病毒感染者进行抗逆转录病毒治疗（ART）， 到2020年，扩大抗逆转录病毒治疗的努力将使超过3000万成年人和超过170万儿童接受抗逆转录病毒治疗，费用几乎为 530亿美元。因此，接受基于蛋白酶抑制剂（PI）的替代ART的患者数量 一线治疗方案失败后的死亡率也将增加。资源有限地区患者接受基于PI的ART的研究 一些国家在治疗12个月和24个月时病毒学失败率很高， 基于PI的治疗方案失败后，会积累耐药突变（DRM）， 治疗方案。资源限制往往限制了收集基因型信息的能力， 与基于耐药性的患者分层相比，药物转换已被证明是成本效益低的 基因型因此，迫切需要为PI失败的患者提供负担得起的HIV基因分型选择， 聚焦基因分型可以通过以下方式显著增强患者护理：1）优化NRTI骨架 仅NRTI耐药的PI方案失败的患者;或2）防止失败的患者转换ART 没有阻力，直到遵守问题得到纠正。阿尔达图的泛简并放大与适应 （PANDAA™）技术是一种新颖的点突变测定法，其使得能够进行廉价且高通量聚焦的点突变测定。 基因型耐药性检测，这种方法可以节省国家ART计划的成本。熊猫 通过去除次级多态性来补偿患者内和患者间的高HIV基因组变异性， 最大限度地减少它们对qPCR灵敏度/特异性的影响，并首次实现qPCR用于HIV基因分型。 可行性研究表明，PANDAA：1）检测NNRTI和NRTI耐药HIV变异体， 灵敏度>99%; 2）不依赖于HIV亚型; 3）可多重同时量化耐药性 在多个基因组位置。Aldatu开创了PANDAA的商业开发，并建立了 一种试剂制剂，其允许在热稳定的样品中产生基于PANDAA的诊断， 准备格式。在这个二期项目中，阿尔达图将把PANDAA技术应用到PANDAA的开发中 PIDR+，一种快速、低成本、热稳定的检测方法，用于检测基于PI的ART失败患者的耐药性 该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 为了实现本文提出的目标，我们将1）实验验证PANDAA试剂的设计，以量化突变 与蛋白酶抑制剂抗性相关，包括10%的病毒准种; 2）建立广泛的， 用于PANDAA PIDR+验证的耐药和药物敏感HIV-1分离株的协作熟练度小组; 3)使用HIV耐药性基因分型的既定性能标准评估PANDAA PIDR+， 在GMP条件下生产PANDAA PIDR+;以及4）验证最终用户，多地点实施 PANDAA PIDR+具有高度可重复性。首款经GMP认证的PANDAA PIDR+检测试剂盒 Aldatu将准备在这个快速增长的诊断市场机会中占据重要份额。