High-content analysis and Kinetic Image Cytometry identify toxicity and epigenetic effects of HIV antiretrovirals on human iPSC-neurons and primary neural precursor cells

Smith, Alyson S.; Ankam, Soneela; Farhy, Chen; Fiengo, Lorenzo; Basa, Ranor; Gordon, Kara L.; Martin, Charles T.; Terskikh, Alexey V.; Jordan‐Sciutto, Kelly L.; Price, Jeffrey H.; McDonough, Patrick M.

doi:10.1016/j.vascn.2022.107157

Cited by 9 publications

(9 citation statements)

References 112 publications

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“…Our observation of ETR-induced synapse loss is in line with the neural toxicity observed by the same group ( Robertson et al, 2012 ). Work from Smith et al has shown that prolonged, 7 days exposures to elvitegravir, DTG/TDF/FTC, EVG/TDF/FTC, and TDF/FTC can reduce synapse density relative to controls in hiPSC-derived neurons but did not observe synapse loss or neuronal toxicity within 24 h ( Smith et al, 2022 ). The use of primary rodent neurons versus hiPSC-derived neurons and the shorter incubation time may account for the lack of synapse loss detected in the assay described here.…”

Section: Discussionmentioning

confidence: 99%

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Antiretroviral drugs from multiple classes induce loss of excitatory synapses between hippocampal neurons in culture

McMullan,

Gansemer,

Thayer

2024

Front. Pharmacol.

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Introduction: Antiretroviral (ARV) drugs have improved prognoses for people living with HIV. However, HIV-associated neurocognitive disorders (HAND) persist despite undetectable viral loads. Some ARVs have been linked to neuropsychiatric effects that may contribute to HAND. Synapse loss correlates with cognitive decline in HAND and synaptic deficits may contribute to the neuropsychiatric effects of ARV drugs.Methods: Using an automated high content assay, rat hippocampal neurons in culture expressing PSD95-eGFP to label glutamatergic synapses and mCherry to fill neuronal structures were imaged before and after treatment with 25 clinically used ARVs.Results and Discussion: At a concentration of 10 μM the protease inhibitors nelfinavir and saquinavir, the non-nucleoside reverse transcriptase inhibitors etravirine and the 8-OH metabolite of efavirenz, the integrase inhibitor bictegravir, and the capsid inhibitor lenacapavir produced synaptic toxicity. Only lenacapavir produced synapse loss at the nanomolar concentrations estimated free in the plasma, although all 4 ARV drugs induced synapse loss at Cmax. Evaluation of combination therapies did not reveal synergistic synaptic toxicity. Synapse loss developed fully by 24 h and persisted for at least 3 days. Bictegravir-induced synapse loss required activation of voltage-gated Ca2+ channels and bictegravir, etravirine, and lenacapavir produced synapse loss by an excitotoxic mechanism. These results indicate that select ARV drugs might contribute to neuropsychiatric effects in combination with drugs that bind serum proteins or in disease states in which synaptic function is altered. The high content imaging assay used here provides an efficient means to evaluate new drugs and drug combinations for potential CNS toxicity.

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Section: Discussionmentioning

confidence: 99%

“…Clearly human neurons might express unique drug sensitivities lacking in synaptic networks derived from rodent neurons. Studies using hiPSC-derived neuron cultures, which have largely intact signaling networks ( Autar et al, 2022 ; Smith et al, 2022 ; Asher et al, 2023 ), could also increase translatability of such research.…”

Section: Discussionmentioning

confidence: 99%

Antiretroviral drugs from multiple classes induce loss of excitatory synapses between hippocampal neurons in culture

McMullan,

Gansemer,

Thayer

2024

Front. Pharmacol.

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“…On the one hand, it could be a search for drugs inducing cancer cell differentiation, or rejuvenating drugs. On the other hand, fluorescence-based screening could be used as a safety test to reveal the hidden epigenetic toxicity of compounds that are under development or already in use [ 99 ]. However, the computation-rich MIEL strategy works with fixed cells, thus missing the dynamic picture of epigenetic remodeling.…”

Section: Discussionmentioning

confidence: 99%

Studying Chromatin Epigenetics with Fluorescence Microscopy

Stepanov

Besedovskaia

Moshareva

et al. 2022

IJMS

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Epigenetic modifications of histones (methylation, acetylation, phosphorylation, etc.) are of great importance in determining the functional state of chromatin. Changes in epigenome underlay all basic biological processes, such as cell division, differentiation, aging, and cancerous transformation. Post-translational histone modifications are mainly studied by immunoprecipitation with high-throughput sequencing (ChIP-Seq). It enables an accurate profiling of target modifications along the genome, but suffers from the high cost of analysis and the inability to work with living cells. Fluorescence microscopy represents an attractive complementary approach to characterize epigenetics. It can be applied to both live and fixed cells, easily compatible with high-throughput screening, and provide access to rich spatial information down to the single cell level. In this review, we discuss various fluorescent probes for histone modification detection. Various types of live-cell imaging epigenetic sensors suitable for conventional as well as super-resolution fluorescence microscopy are described. We also focus on problems and future perspectives in the development of fluorescent probes for epigenetics.

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“…The introduction of highly active antiretroviral therapy (HAART) led to a remarkable reduction of HIV associated dementia (HAND) and control of HIV replication in the CNS [191,192]. However, in the post-HAART period, there has been a rise in milder forms of dementia possibly associated to toxicity related to the continuous use of ART [193][194][195][196][197]. Even in the context of plasma viral suppression, some PLWH experience CSF/plasma discordance, meaning that HIV RNA levels are higher in CSF than in plasma, raising concerns about drug penetrance in the brain and CSF viral escape [176,[198][199][200][201][202].…”

Section: Hiv Infection and Maintenance Of Reservoirs In The Brainmentioning

confidence: 99%

HIV Latency in Myeloid Cells: Challenges for a Cure

et al. 2022

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The use of antiretroviral therapy (ART) for Human Immunodeficiency Virus (HIV) treatment has been highly successful in controlling plasma viremia to undetectable levels. However, a complete cure for HIV is hindered by the presence of replication-competent HIV, integrated in the host genome, that can persist long term in a resting state called viral latency. Resting memory CD4+ T cells are considered the biggest reservoir of persistent HIV infection and are often studied exclusively as the main target for an HIV cure. However, other cell types, such as circulating monocytes and tissue-resident macrophages, can harbor integrated, replication-competent HIV. To develop a cure for HIV, focus is needed not only on the T cell compartment, but also on these myeloid reservoirs of persistent HIV infection. In this review, we summarize their importance when designing HIV cure strategies and challenges associated to their identification and specific targeting by the “shock and kill” approach.

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High-content analysis and Kinetic Image Cytometry identify toxicity and epigenetic effects of HIV antiretrovirals on human iPSC-neurons and primary neural precursor cells

Cited by 9 publications

References 112 publications

Antiretroviral drugs from multiple classes induce loss of excitatory synapses between hippocampal neurons in culture

Antiretroviral drugs from multiple classes induce loss of excitatory synapses between hippocampal neurons in culture

Studying Chromatin Epigenetics with Fluorescence Microscopy

HIV Latency in Myeloid Cells: Challenges for a Cure

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