| Publication Abstract Display | | Type: Poster | | Title: Neurotoxicity screening of antiretroviral drugs with human iPSC-derived neurons. | | Authors: Hinckley S, Sherman S, Brookie M, Momper J, Ma Q, Letendre S, Ellis R, Bang A | | Date: 02-22-2016 | | Abstract:While antiretroviral therapy (ART) has become increasingly effective and well-tolerated, there remains a patient subset that experience central nervous system (CNS) side effects, and neurocognitive performance may actually improve following change in specific ART agents. Understanding the risk associated with these drugs will allow informed selection of optimal therapy. Using human induced pluripotent stem cell-derived neurons (hiPSC), we screened 10 ART drugs and generated a neurotoxicity profile based on mitochondrial membrane potential, reactive oxygen species, cell health and neurite growth.
hiPSC cortical neurons (Cellular Dynamics International ) were treated in dose response with ABC,ATV,COBI,DRV,DTG,EFV,EVG,RPV,RTV,TDF. Neurons were assayed for mitochondrial function and neurite growth using image based high content, high throughput screening. Statistical significance was determined as Z-score greater than 2 compared with vehicle control mean across replicate wells.
The majority of tested drugs demonstrated neurotoxicity: 7 caused mitochondrial toxicity and 3 affected neurite growth. Non-nucleoside reverse transcriptase inhibitor, protease inhibitor (PI) and pharmacoenhancer drugs exhibited the highest degree of mitochondrial toxicity. Only EFV was overtly cytotoxic suggesting mitochondrial dysfunction was a primary target, not a side effect of cell death. After 3 days exposure to ART drugs, morphology and cell health was assessed. DTG and EFV resulted in minor but significant neurite growth. RPV was cytotoxic at high dose but caused neurite inhibition at lower doses. Of note, neurite outgrowth or inhibition could contribute to CNS pathology. At high doses, COBI, EVG, RPV and EFV caused neurite inhibition coincident with cytoxicity suggesting the morphological effect was secondary to cell death. While PIs influenced mitochondrial function, no effect was found on morphology. In contrast, COBI, EVG, RPV and EFV presented a distinct profile with mitochondrial dysfunction followed by changes in morphology and/or cytoxicity.
We characterized toxicity of 10 ART drugs and linked the majority with impacts either on mitochondrial function, neurite growth, or both. Although immature, hiPSC neurons are human and scalable for drug screening. Further studies are needed to determine whether our in vitro assays reflect neurotoxicity in vivo, but our results suggest that increased drug concentrations in the CNS could have adverse clinical consequences. |
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