Alcohol-and-HIV-Induced Lysosomal Dysfunction Regulates Extracellular Vesicles Secretion in Vitro and in Liver-Humanized Mice

Authors

Raghubendra Singh Dagur, Veterans Affairs Nebraska-Western Iowa Health Care System
Moses New-Aaron, Veterans Affairs Nebraska-Western Iowa Health Care System
Murali Ganesan, Veterans Affairs Nebraska-Western Iowa Health Care System
Weimin Wang, University of Nebraska Medical CenterFollow
Svetlana Romanova, University of Nebraska Medical CenterFollow
Srivatsan Kidambi, University of NebraskaFollow
Kusum K. Kharbanda, Veterans Affairs Nebraska-Western Iowa Health Care System
Larisa Y. Poluektova, University of Nebraska Medical CenterFollow
Natalia A. Osna, Veterans Affairs Nebraska-Western Iowa Health Care System

ORCID IDs

Raghubendra Singh Dagur

Moses New-Aaron

Srivatsan Kidambi

Kusum K. Kharbanda

Larisa Y. Poluektova

Natalia A. Osna

Date of this Version

1-5-2021

Document Type

Article

Citation

Dagur, R.S.; New-Aaron, M.; Ganesan, M.; Wang, W.; Romanova, S.; Kidambi, S.; Kharbanda, K.K.; Poluektova, L.Y.; Osna, N.A. Alcohol-and-HIV-Induced Lysosomal Dysfunction Regulates Extracellular Vesicles Secretion in Vitro and in Liver-Humanized Mice. Biology 2021, 10, 29. https://doi.org/10.3390/ biology10010029

Comments

CC-BY

Abstract

Background: Alcohol abuse is common in people living with HIV-1 and dramatically enhances the severity of HIV-induced liver damage by inducing oxidative stress and lysosomal dysfunction in the liver cells. We hypothesize that the increased release of extracellular vesicles (EVs) in hepatocytes and liver humanized mouse model is linked to lysosome dysfunction. Methods: The study was performed on primary human hepatocytes and human hepatoma RLW_XP-GFP (Huh 7.5 cells stably transfected with CYP2E1 and XPack-GFP) cells and validated on ethanol-fed liver humanized fumarylacetoacetate hydrolase (Fah)^-/-, Rag2^-/-, common cytokine receptor gamma chain knockout (FRG-KO) mice. Cells and mice were infected with HIV-1_ADA virus. Results: We observed an increase in the secretion of EVs associated with a decrease in lysosomal activity and expression of lysosomal-associated membrane protein 1. Next-generation RNA sequencing of primary human hepatocytes revealed 63 differentially expressed genes, with 13 downregulated and 50 upregulated genes in the alcohol–HIV-treated group. Upstream regulator analysis of differentially expressed genes through Ingenuity Pathway Analysis identified transcriptional regulators affecting downstream genes associated with increased oxidative stress, lysosomal associated disease, and function and EVs biogenesis. Our in vitro findings were corroborated by in vivo studies on human hepatocyte transplanted humanized mice, indicating that intensive EVs’ generation by human hepatocytes and their secretion to serum was associated with increased oxidative stress and reduction in lysosomal activities triggered by HIV infection and ethanol diet. Conclusion: HIV-and-ethanol-metabolism induced EVs release is tightly controlled by lysosome status in hepatocytes and participates in the development of double-insult-induced liver injury