Mitochondrial Dysfunction in HAART: Point of Care Tests

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 gt90% 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.