High-content analysis and Kinetic Image Cytometry identify toxicity and epigenetic effects of HIV antiretrovirals on human iPSC-neurons and primary neural precursor cells.
High-content analysis and Kinetic Image Cytometry identify toxicity and epigenetic effects of HIV antiretrovirals on human iPSC-neurons and primary neural precursor cells.
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DOI：
10.1016/j.vascn.2022.107157
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发表时间：
2022-03
影响因子：
1.9
通讯作者：
McDonough PM
中科院分区：
文献类型：
--
作者：
Smith AS;Ankam S;Farhy C;Fiengo L;Basa RCB;Gordon KL;Martin CT;Terskikh AV;Jordan-Sciutto KL;Price JH;McDonough PM
尽管由于联合抗逆转录病毒治疗（cART）而抑制了病毒，但HIV相关的神经认知障碍（HAND）继续影响一半的HIV感染者，这表明某些抗逆转录病毒药物（ARV）可能导致HAND。我们研究了核苷/核苷酸逆转录酶抑制剂富马酸替诺福韦酯（TDF）和恩曲他滨（FTC）以及整合酶抑制剂度鲁特韦（DTG）和埃替拉韦（EVG）对多巴胺能神经元活力、结构和功能的影响。（一种参与认知的中枢神经系统神经元亚型），来源于人类诱导多能干细胞（hiPSC-神经元）和负责神经发生的原代人神经前体细胞（hNPC）。使用自动数字显微镜和图像分析（高含量分析，HCA），我们发现DTG、EVG和TDF在处理7天后降低hiPSC神经元活力、神经突和突触。使用动力学图像细胞计数法（KIC）分析hiPSC-神经元钙活性表明，DTG和EVG还降低了细胞内钙瞬变的频率和幅度。长期接触抗逆转录病毒药物和同时接触多种抗逆转录病毒药物增加了这些神经毒性作用的程度。使用显微镜成像表观遗传景观（MIEL）分析，我们发现，TDF降低hNPC的活力和改变的组蛋白修饰，调节染色质包装的分布，这表明TDF可能会减少神经祖细胞池重要的中枢神经系统的发展和维持成人的认知。这项研究建立了人类临床前试验，可以筛选潜在的抗逆转录病毒药物的中枢神经系统毒性，以开发更安全的cART方案和HAND疗法。
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 disoproxil 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.
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影响因子：
10.5
作者：
Dahabieh MS;Battivelli E;Verdin E
通讯作者：
Verdin E
DOI：
10.1056/nejmoa1600693
发表时间：
2016-09-01
期刊：
The New England journal of medicine
影响因子：
--
作者：
Cohen MS;Chen YQ;McCauley M;Gamble T;Hosseinipour MC;Kumarasamy N;Hakim JG;Kumwenda J;Grinsztejn B;Pilotto JH;Godbole SV;Chariyalertsak S;Santos BR;Mayer KH;Hoffman IF;Eshleman SH;Piwowar-Manning E;Cottle L;Zhang XC;Makhema J;Mills LA;Panchia R;Faesen S;Eron J;Gallant J;Havlir D;Swindells S;Elharrar V;Burns D;Taha TE;Nielsen-Saines K;Celentano DD;Essex M;Hudelson SE;Redd AD;Fleming TR;HPTN 052 Study Team
通讯作者：
HPTN 052 Study Team
影响因子：
7.7
作者：
Farhy, Chen;Hariharan, Santosh;Terskikh, Alexey V.
通讯作者：
Terskikh, Alexey V.
影响因子：
6.2
作者：
Chang, L.;Yakupov, R.;Ernst, T.
通讯作者：
Ernst, T.
影响因子：
4.3
作者：
Demir M;Laywell ED
通讯作者：
Laywell ED