Department of Genomic Medicine, Lerner Research Institute, Cleveland Clinic, Cleveland, OH 44195, USA.
Center for Molecular Medicine and Genetics, Wayne State University, Detroit, MI 48202, USA.
Int J Mol Sci. 2023 Jul 31;24(15):12242. doi: 10.3390/ijms241512242.
HIV-associated cognitive dysfunction during combination antiretroviral therapy (cART) involves mitochondrial dysfunction, but the impact of contemporary cART on chronic metabolic changes in the brain and in latent HIV infection is unclear. We interrogated mitochondrial function in a human microglia (hμglia) cell line harboring inducible HIV provirus and in SH-SY5Y cells after exposure to individual antiretroviral drugs or cART, using the MitoStress assay. cART-induced changes in protein expression, reactive oxygen species (ROS) production, mitochondrial DNA copy number, and cellular iron were also explored. Finally, we evaluated the ability of ROS scavengers or plasmid-mediated overexpression of the antioxidant iron-binding protein, Fth1, to reverse mitochondrial defects. Contemporary antiretroviral drugs, particularly bictegravir, depressed multiple facets of mitochondrial function by 20-30%, with the most pronounced effects in latently infected HIV+ hμglia and SH-SY5Y cells. Latently HIV-infected hμglia exhibited upregulated glycolysis. Increases in total and/or mitochondrial ROS, mitochondrial DNA copy number, and cellular iron accompanied mitochondrial defects in hμglia and SH-SY5Y cells. In SH-SY5Y cells, cART reduced mitochondrial iron-sulfur-cluster-containing supercomplex and subunit expression and increased Nox2 expression. Fth1 overexpression or pre-treatment with N-acetylcysteine prevented cART-induced mitochondrial dysfunction. Contemporary cART impairs mitochondrial bioenergetics in hμglia and SH-SY5Y cells, partly through cellular iron accumulation; some effects differ by HIV latency.
HIV 相关认知功能障碍在联合抗逆转录病毒治疗(cART)期间涉及线粒体功能障碍,但当代 cART 对大脑慢性代谢变化和潜伏性 HIV 感染的影响尚不清楚。我们使用 MitoStress 检测法,在携带诱导型 HIV 前病毒的人小胶质细胞(hμglia)细胞系和暴露于单个抗逆转录病毒药物或 cART 的 SH-SY5Y 细胞中检测了线粒体功能。还研究了 cART 诱导的蛋白质表达、活性氧(ROS)产生、线粒体 DNA 拷贝数和细胞内铁的变化。最后,我们评估了 ROS 清除剂或质粒介导的抗氧化铁结合蛋白 Fth1 的过表达逆转线粒体缺陷的能力。当代抗逆转录病毒药物,特别是比克替拉韦,使线粒体功能的多个方面降低了 20-30%,对潜伏感染的 HIV+hμglia 和 SH-SY5Y 细胞的影响最为明显。潜伏感染的 HIV hμglia 表现出糖酵解增加。总 ROS 和/或线粒体 ROS、线粒体 DNA 拷贝数和细胞内铁的增加伴随着 hμglia 和 SH-SY5Y 细胞中线粒体缺陷。在 SH-SY5Y 细胞中,cART 降低了线粒体铁硫簇结合的超级复合物和亚基表达,并增加了 Nox2 表达。Fth1 过表达或 N-乙酰半胱氨酸预处理可预防 cART 诱导的线粒体功能障碍。当代 cART 损害了 hμglia 和 SH-SY5Y 细胞中的线粒体生物能,部分原因是细胞内铁积累;一些影响因 HIV 潜伏期而异。