HIV-1 Tat endocytosis and retention in endolysosomes affects HIV-1 Tat-induced LTR transactivation in astrocytes

• Published in: The FASEB journal Vol. 36; no. 3; p. e22184
• Main Authors: Khan, Nabab, Halcrow, Peter W, Afghah, Zahra, Baral, Aparajita, Geiger, Jonathan D, Chen, Xuesong
• Format: Journal Article
• Language: English
• Published: United States 01.03.2022
• Subjects: Astrocytes - virology; Cell Line, Tumor; Endocytosis - genetics; Endosomes - genetics; Gene Expression Regulation, Viral - genetics; HIV Infections - genetics; HIV Infections - virology; HIV Long Terminal Repeat - genetics; HIV-1 - genetics; Humans; Lysosomes - genetics; Promoter Regions, Genetic - genetics; tat Gene Products, Human Immunodeficiency Virus - genetics; Transcriptional Activation - genetics; Virus Latency - genetics; Virus Replication - genetics; endolysosomes; HIV-1 latency; HIV-1 Tat; astrocytes; HIV-1 LTR transactivation
• ISSN: 1530-6860
• DOI: 10.1096/fj.202101722R

The presence of latent HIV-1 reservoirs in the periphery and brain represents a major obstacle to curing HIV-1 infection. As an essential protein for HIV-1 viral replication, HIV-1 Tat, mostly intracellular, has been implicated in latent HIV-1 infection. From HIV-1 infected cells, HIV-1 Tat is actively secreted and bystander cells uptake the released Tat whereupon it is endocytosed and internalized into endolysosomes. However, to activate the HIV-1 LTR promoter and increase HIV-1 replication, HIV-1 Tat must first escape from the endolysosomes and then enter the nucleus. Here, we tested the hypothesis that HIV-1 Tat can accumulate in endolysosomes and contribute to the activation of latent HIV-1 in astrocytes. Using U87MG astrocytoma cells expressing HIV-1 LTR-driven luciferase and primary human astrocytes we found that exogenous HIV-1 Tat enters endolysosomes, resides in endolysosomes for extended periods of time, and induces endolysosome de-acidification as well as enlargement. The weak base chloroquine promoted the release of HIV-1 Tat from endolysosomes and induced HIV-1 LTR transactivation. Similar results were observed by activating endolysosome Toll-like receptor 3 (TLR3) and TLR7/8. Conversely, pharmacological block of TLRs and knocking down expression levels of TLR3 and TLR7, but not TLR8, prevented endolysosome leakage and attenuated HIV-1 Tat-mediated HIV-1 LTR transactivation. Our findings suggest that HIV-1 Tat accumulation in endolysosomes may play an important role in controlling HIV-1 transactivation.