Alcohol-and-HIV-Induced Lysosomal Dysfunction Regulates Extracellular Vesicles Secretion in Vitro and in Liver-Humanized Mice.
Alcohol-and-HIV-Induced Lysosomal Dysfunction Regulates Extracellular Vesicles Secretion in Vitro and in Liver-Humanized Mice.
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DOI：
10.3390/biology10010029
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发表时间：
2021-01-05
期刊：
影响因子：
4.2
通讯作者：
Osna NA
中科院分区：
文献类型：
--
作者：
Dagur RS;New-Aaron M;Ganesan M;Wang W;Romanova S;Kidambi S;Kharbanda KK;Poluektova LY;Osna NA
饮酒会增加艾滋病毒感染者的肝损伤;然而，其机制仍然难以捉摸。通过利用酒精（乙醇）暴露的HIV感染的人肝细胞肝细胞和乙醇喂养的肝脏人源化小鼠模型（免疫缺陷小鼠肝脏含有人肝细胞），我们表明酒精和HIV的组合增强了氧化应激并损害了溶酶体活性，这反过来又刺激了致病性纳米细胞外囊泡。这些囊泡非常重要，因为操纵它们的数量和内容物将消除感染在其他细胞类型中的传播。背景资料：酒精滥用在HIV-1感染者中很常见，并通过诱导肝细胞中的氧化应激和溶酶体功能障碍显著增强HIV诱导的肝损伤的严重程度。我们假设，细胞外囊泡（EV）在肝细胞和肝脏人源化小鼠模型中的释放增加与溶酶体功能障碍有关。研究方法：本研究在原代人肝细胞和人肝癌RLWXP-GFP（经CYP 2 E1和XPack-GFP稳定转染的Huh 7.5细胞）细胞中进行，并在乙醇喂养的肝脏人源化延胡索酰乙酰乙酸水解酶（Fah）-/-、Rag 2-/-、常见细胞因子受体γ链敲除（FRG-KO）小鼠中进行验证。用HIV-1ADA病毒感染细胞和小鼠。结果如下：我们观察到与溶酶体活性和溶酶体相关膜蛋白1表达降低相关的EV分泌增加。原代人肝细胞的下一代RNA测序揭示了63个差异表达的基因，其中13个下调和50个上调基因在酒精-HIV治疗组中。通过免疫途径分析对差异表达基因的上游调节因子进行分析，鉴定了影响下游基因的转录调节因子，这些基因与氧化应激增加、溶酶体相关疾病和功能以及EV生物发生相关。我们的体外研究结果得到了人肝细胞移植人源化小鼠体内研究的证实，表明人肝细胞产生的大量EV及其分泌到血清中与HIV感染和乙醇饮食引发的氧化应激增加和溶酶体活性降低有关。结论：HIV和乙醇代谢诱导的EV释放受肝细胞中溶酶体状态的严格控制，并参与双重损伤诱导的肝损伤的发展。
Consumption of alcohol increases liver damage in HIV people; however, the mechanisms remain elusive. By utilizing alcohol (ethanol) exposed HIV-infected human hepatocytes liver cells and ethanol-fed liver humanized mouse model (immunodeficient mice livers harbor human hepatocytes), we show that the combination of alcohol and HIV enhances oxidative stress and impairs lysosome activity, which in turn stimulates pathogenic nanosized extracellular vesicles. These vesicles are of great importance, as manipulating their numbers and contents will eliminate the spread of infection in other cell types. 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 RLWXP-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-1ADA 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.
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影响因子：
46.9
作者：
Azuma, Hisaya;Paulk, Nicole;Grompe, Markus
通讯作者：
Grompe, Markus
影响因子：
16
作者：
Lötvall J;Hill AF;Hochberg F;Buzás EI;Di Vizio D;Gardiner C;Gho YS;Kurochkin IV;Mathivanan S;Quesenberry P;Sahoo S;Tahara H;Wauben MH;Witwer KW;Théry C
通讯作者：
Théry C
DOI：
10.1097/qai.0000000000000756
发表时间：
2015-11-01
期刊：
Journal of acquired immune deficiency syndromes (1999)
影响因子：
--
作者：
Hubert A;Subra C;Jenabian MA;Tremblay Labrecque PF;Tremblay C;Laffont B;Provost P;Routy JP;Gilbert C
通讯作者：
Gilbert C
影响因子：
14.9
作者：
Babicki S;Arndt D;Marcu A;Liang Y;Grant JR;Maciejewski A;Wishart DS
通讯作者：
Wishart DS
影响因子：
6.1
作者：
Godoy, Patricio;Hewitt, Nicola J.;Albrecht, Ute;Andersen, Melvin E.;Ansari, Nariman;Bhattacharya, Sudin;Bode, Johannes Georg;Bolleyn, Jennifer;Borner, Christoph;Boettger, Jan;Braeuning, Albert;Budinsky, Robert A.;Burkhardt, Britta;Cameron, Neil R.;Camussi, Giovanni;Cho, Chong-Su;Choi, Yun-Jaie;Rowlands, J. Craig;Dahmen, Uta;Damm, Georg;Dirsch, Olaf;Teresa Donato, Maria;Dong, Jian;Dooley, Steven;Drasdo, Dirk;Eakins, Rowena;Ferreira, Karine Sa;Fonsato, Valentina;Fraczek, Joanna;Gebhardt, Rolf;Gibson, Andrew;Glanemann, Matthias;Goldring, Chris E. P.;Jose Gomez-Lechon, Maria;Groothuis, Geny M. M.;Gustavsson, Lena;Guyot, Christelle;Hallifax, David;Hammad, Seddik;Hayward, Adam;Haeussinger, Dieter;Hellerbrand, Claus;Hewitt, Philip;Hoehme, Stefan;Holzhuetter, Hermann-Georg;Houston, J. Brian;Hrach, Jens;Ito, Kiyomi;Jaeschke, Hartmut;Keitel, Verena;Kelm, Jens M.;Park, B. Kevin;Kordes, Claus;Kullak-Ublick, Gerd A.;LeCluyse, Edward L.;Lu, Peng;Luebke-Wheeler, Jennifer;Lutz, Anna;Maltman, Daniel J.;Matz-Soja, Madlen;McMullen, Patrick;Merfort, Irmgard;Messner, Simon;Meyer, Christoph;Mwinyi, Jessica;Naisbitt, Dean J.;Nussler, Andreas K.;Olinga, Peter;Pampaloni, Francesco;Pi, Jingbo;Pluta, Linda;Przyborski, Stefan A.;Ramachandran, Anup;Rogiers, Vera;Rowe, Cliff;Schelcher, Celine;Schmich, Kathrin;Schwarz, Michael;Singh, Bijay;Stelzer, Ernst H. K.;Stieger, Bruno;Stoeber, Regina;Sugiyama, Yuichi;Tetta, Ciro;Thasler, Wolfgang E.;Vanhaecke, Tamara;Vinken, Mathieu;Weiss, Thomas S.;Widera, Agata;Woods, Courtney G.;Xu, Jinghai James;Yarborough, Kathy M.;Hengstler, Jan G.
通讯作者：
Hengstler, Jan G.