PT  - JOURNAL ARTICLE
AU  - Alyson S. Smith
AU  - Soneela Ankam
AU  - Chen Farhy
AU  - Lorenzo Fiengo
AU  - Ranor C.B. Basa
AU  - Kara L. Gordon
AU  - Charles T. Martin
AU  - Alexey V. Terskikh
AU  - Kelly L. Jordan-Sciutto
AU  - Jeffrey H. Price
AU  - Patrick M. McDonough
TI  - High-content analysis and Kinetic Image Cytometry identify toxic and epigenotoxic effects of HIV antiretrovirals on human iPSC-neurons and primary neural precursor cells
AID  - 10.1101/2020.09.05.284422
DP  - 2021 Jan 01
TA  - bioRxiv
PG  - 2020.09.05.284422
4099  - http://biorxiv.org/content/early/2021/01/11/2020.09.05.284422.short
4100  - http://biorxiv.org/content/early/2021/01/11/2020.09.05.284422.full
AB  - Despite viral suppression due to combination antiretroviral therapy (cART), HIV-associated neurocognitive disorders (HAND) continue to affect half of people with HIV, suggesting that certain antiretrovirals (ARVs) may contribute to HAND. We examined the effects of nucleoside/nucleotide reverse transcriptase inhibitors tenofovir disproxil fumarate (TDF) and emtricitabine (FTC) and the integrase inhibitors dolutegravir (DTG) and elvitegravir (EVG) on viability, structure, and function of glutamatergic neurons (a subtype of CNS neuron involved in cognition) derived from human induced pluripotent stem cells (hiPSC-neurons), and primary human neural precursor cells (hNPCs), which are responsible for neurogenesis. Using automated digital microscopy and image analysis (high content analysis, HCA), we found that DTG, EVG, and TDF decreased hiPSC-neuron viability, neurites, and synapses after seven days of treatment. Analysis of hiPSC-neuron calcium activity using Kinetic Image Cytometry (KIC) demonstrated that DTG and EVG also decreased the frequency and magnitude of intracellular calcium transients. Longer ARV exposures and simultaneous exposure to multiple ARVs increased the magnitude of these neurotoxic effects. Using the Microscopic Imaging of Epigenetic Landscapes (MIEL) assay, we found that TDF decreased hNPC viability and changed the distribution of histone modifications that regulate chromatin packing, suggesting that TDF may reduce neuroprogenitor pools important for CNS development and maintenance of cognition in adults. This study establishes human preclinical assays that can screen potential ARVs for CNS toxicity to develop safer cART regimens and HAND therapeutics.Competing Interest StatementThe authors have declared no competing interest.