Mitochondrial Dysfunction in HAART: Point of Care Tests

描述(申请人提供)：用于治疗艾滋病毒/艾滋病的高效抗逆转录病毒疗法(HAART)有严重的副作用，包括线粒体毒性引起的代谢并发症，可能危及生命，并限制有效性和患者依从性。不良代谢影响现在由熟练的临床医生管理，他们密切监测患者的临床症状，并相应地调整治疗方法。然而，改进早期毒性检测可以帮助医生在对患者风险较小的情况下管理副作用，并避免出现临床症状，其中一些症状是持续的，而且只有部分可逆。线粒体氧化磷酸化(OXPHOS)蛋白水平是HAART毒性的早期生物标志物，因为OXPHOS通常提供90%的细胞能量。在第一阶段开发的一套试纸免疫分析测试已经用于临床研究，表明外周脂肪和外周血单核细胞(PBMC)中的OXPHOS缺陷与HAART诱导的脂肪萎缩相关。这些检测是高度定量的，但简单快速，因为基本平台类似于许多应用程序中使用的，如家用验孕试剂盒。第二阶段的工作将改进测试，增加内部加载控制，以便于比率读出，并将试纸合并到坚固的一次性盒式磁带中。这些改进将简化样品处理需求和数据分析，允许在护理点应用。它还表明，这种测试可以用于容易采集的组织，包括脸颊拭子、指刺全血和尿液沉渣。第二阶段的工作包括临床研究，以确定这些容易取样的组织中的OXPHOS缺陷是否与HAART(脂肪萎缩和周围神经病变)的不利代谢影响的临床症状相关，如果是，是否如我们的初步研究中强烈建议的那样，OXPHOS缺陷是否先于症状出现。确凿的结果将极大地支持这些测试的实用性，以帮助指导治疗，将HAART的长期副作用降至最低。此外，由于线粒体能量产生对细胞功能至关重要，OXPHOS障碍也与许多其他情况有关。第一代氧磷酸盐试纸已经用于遗传性线粒体疾病、神经退行性疾病(帕金森病和弗里德里希S共济失调)以及其他代谢疾病的研究，如败血症和癌症。这些测试在上述每个领域和其他领域都有潜在的诊断/治疗作用。这些测试现在还被用于制药合同药物安全筛选，以确定新治疗药物的有丝分裂毒性副作用，不仅包括新的抗逆转录病毒药物，而且还有许多其他药物，如抗生素和抗癌药物，对这些药物来说，有丝分裂毒性是一个既定的风险。因此，这些测试将在第三阶段商业化和销售：1)帮助指导HAART和其他代谢状况的诊断/治疗辅助工具，2)研究HAART和其他线粒体疾病不利代谢影响的分子病理学的研究工具，以及3)用于药物安全性筛选的有丝分裂毒性和线粒体生物发生分析。我们正在进行简单、廉价的试纸测试，可以对容易获得的样本(脸颊拭子、指尖采血和尿液)进行测试，以监测HAART的不良反应，并帮助指导治疗，将毒副作用降至最低。因为这些测试有助于测量患者S分解和利用食物中能量的能力，它们也可以用来监测许多其他疾病中已知的类似干扰，包括帕金森氏病、癌症、败血症和遗传病，还可以用来筛选类似毒性作用的新治疗药物，避免在患者身上使用它们。

DESCRIPTION (provided by applicant): Highly Active Anti-Retroviral Therapy (HAART) used to treat HIV/AIDS has serious side-effects, including metabolic complications from mitochondrial toxicities that can be life-threatening, and limit effectiveness and patient compliance. Adverse metabolic effects are now managed by adept clinicians who monitor patients closely for clinical symptoms and adjust therapies accordingly. However, improved earlier detection of toxicities could help physicians manage side effects with less risk to patients and avoid onset of clinical symptoms, some of which are persistent and only partially reversible. Mitochondrial Oxidative Phosphorylation (OXPHOS) protein levels are good candidates for early biomarkers of HAART toxicity as OXPHOS normally makes >90% of cellular energy. A set of dipstick immunoassay tests developed in Phase I have been used in clinical studies to show that OXPHOS deficits in both peripheral fat and Peripheral Blood Mononucleated Cells (PBMCs) correlate with HAART-induced lipoatrophy. The tests are highly quantitative, yet simple and rapid as the basic platform is similar to that used in many applications such as home-use pregnancy test kits. Phase II work will improve the tests by adding an internal loading control to facilitate ratiometric readout, and incorporating the dipsticks into robust single-use cassettes. The improvements will streamline sample processing needs and data analysis, allowing application at point of care. It has also been shown that the tests can be used with easily sampled tissues, including cheek swabs, fingerprick whole blood and urinary sediment. Phase II work includes clinical research to determine if OXPHOS deficits in these easily sampled tissues correlate with clinical symptoms of adverse metabolic effects of HAART (lipoatrophy and peripheral neuropathy) and if so, whether or not OXPHOS deficits precede onset of symptoms as strongly suggested in our pilot study. Confirmatory results would greatly support utility of the tests to help guide therapy to minimize long-term side effects of HAART. Moreover, because mitochondrial energy production is essential for cellular function, OXPHOS disorders are involved in many other conditions. The 1st generation OXPHOS dipstick tests are already being used in studies of inherited mitochondrial diseases, neurodegenerative disorders (PD and Friedreich s Ataxia), and other metabolic disorders such as sepsis and cancer. The tests have potential diagnostic/theranostic utility in each of these and other areas. The tests are also now being used in Pharmaceutical contract drug safety screening to identify mitotoxic side effects of new therapeutic drugs, including not just new antiretrovirals, but many other drugs such as antibiotics and anti-cancer drugs, for which mitotoxicity is an established risk. Therefore, the tests will be commercialized and marketed in Phase III as: 1) diagnostic/theranostic aids to help guide HAART and other metabolic conditions, 2) research tools to study the molecular pathology of the adverse metabolic effects of HAART and other mitochondrial disorders, and 3) mitotoxicity and mitochondrial biogenesis assays for Pharmaceutical drug safety screening.RELEVANCE: Highly Active Anti-Retroviral Therapy (HAART) used to treat HIV/AIDS has serious adverse side-effects that can be life-threatening and limit effectiveness of treatment. We are making simple, inexpensive dipstick tests that can be used with easily obtained samples (cheek swab, fingerprick blood and urine) to monitor the adverse effects of HAART and help guide therapy to minimize toxic side-effects. Because the tests help measure a patient s ability to break down and use energy in food, they can also be used to monitor similar disturbances known to occur in many other conditions, including Parkinsons disease, cancer, sepsis and inherited diseases, and also to screen new therapeutic drugs for similar toxic effects and avoid their use in patients.