CAGE-Seq Reveals that HIV-1 Latent Infection Does Not Trigger Unique Cellular Responses in a Jurkat T Cell Model
CAGE-Seq Reveals that HIV-1 Latent Infection Does Not Trigger Unique Cellular Responses in a Jurkat T Cell Model
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CAGE-Seq 揭示 HIV-1 潜伏感染不会在 Jurkat T 细胞模型中引发独特的细胞反应
DOI：
10.1128/jvi.02394-20
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发表时间：
2021
影响因子：
5.4
通讯作者：
Verdin
中科院分区：
文献类型：
--
作者：
Matsui Hiroyuki;Shirakawa Kotaro;Konishi Yoshinobu;Hirabayashi Shigeki;Sarca Anamaria Daniela;Fukuda Hirofumi;Nomura Ryosuke;Stanford Emani;Horisawa Yoshihito;Kazuma Yasuhiro;Matsumoto Tadahiko;Yamazaki Hiroyuki;Murakawa Yasuhiro;Battivelli Emilie;Verdin
HIV-1的治疗目前因我们无法特异性识别和靶向潜伏感染细胞而停滞不前。HIV-1病毒RNA/DNA或病毒蛋白被细胞机制识别，并在病毒产生细胞中诱导干扰素应答，但潜伏感染细胞的变化仍不清楚。HIVGKO包含HIV-1启动子下的绿色荧光蛋白（GFP）报告基因和内部延伸因子α（EF 1 α）启动子下的单体Kusabira橙子2（mKO 2）报告基因。这种病毒构建体能够直接鉴定生产性和潜伏性HIV-1感染的细胞。在这项研究中，我们的目的是确定特定的细胞转录反应触发的HIV-1的进入和整合使用帽分析基因表达（CAGE）。我们在未感染和潜伏和生产性感染的细胞中对CAGE标签进行深度测序，并比较其差异表达的转录起始位点（TSS）谱。与未感染的细胞或潜伏感染的细胞相比，病毒产生细胞具有与T细胞活化和凋亡相关的差异表达的TSS。令人惊讶的是，与未感染的细胞相比，潜伏感染的细胞仅具有33个差异表达的TSS。其中，SPP 1和APOE在潜伏感染的细胞中下调。在Jurkat T细胞中SPP 1或APOE敲低增加了对HIVGKO感染的易感性，表明它们具有抗病毒特性。磷脂酰肌醇3-激酶（PI 3 K）/哺乳动物雷帕霉素靶蛋白（mTOR）通路的组分MLST 8、4 EBP和RPS 6在生产性感染细胞中是显著的TSS，与潜伏感染细胞相比，S6激酶（S6 K）磷酸化增加，表明mTOR通路活性在建立潜伏库中起作用。这些发现表明，HIV-1进入和整合不触发独特的转录反应时，感染成为latent.IMPORTANCELatent HIV-1感染是建立早在第一次病毒暴露，仍然是最重要的障碍，在获得治愈HIV-1感染。在这里，我们使用基因表达的帽分析（CAGE）来比较潜伏感染细胞与未感染或生产性感染细胞的转录景观。我们发现，潜伏感染的细胞和未感染的细胞表现出非常相似的转录谱。我们的数据表明，当感染仍然潜伏时，T细胞不能识别进入的病毒成分或整合的HIV-1基因组。这些发现应该指导未来的研究扩大我们的方法来识别和靶向潜伏的HIV-1感染细胞。
The cure for HIV-1 is currently stalled by our inability to specifically identify and target latently infected cells. HIV-1 viral RNA/DNA or viral proteins are recognized by cellular mechanisms and induce interferon responses in virus-producing cells, but changes in latently infected cells remain unknown. HIVGKOcontains a green fluorescent protein (GFP) reporter under the HIV-1 promoter and a monomeric Kusabira orange 2 (mKO2) reporter under the internal elongation factor alpha (EF1α) promoter. This viral construct enables direct identification of both productively and latently HIV-1-infected cells. In this study, we aim to identify specific cellular transcriptional responses triggered by HIV-1 entry and integration using cap analysis of gene expression (CAGE). We deep sequenced CAGE tags in non-infected and latently and productively infected cells and compared their differentially expressed transcription start site (TSS) profiles. Virus-producing cells had differentially expressed TSSs related to T-cell activation and apoptosis compared to those of non-infected cells or latently infected cells. Surprisingly, latently infected cells had only 33 differentially expressed TSSs compared to those of non-infected cells. Among these, SPP1 and APOE were downregulated in latently infected cells. SPP1 or APOE knockdown in Jurkat T cells increased susceptibility to HIVGKOinfection, suggesting that they have antiviral properties. Components of the phosphatidylinositol 3-kinase (PI3K)/mammalian target of rapamycin (mTOR) pathway, MLST8, 4EBP, and RPS6, were significant TSSs in productively infected cells, and S6 kinase (S6K) phosphorylation was increased compared to that in latently infected cells, suggesting that mTOR pathway activity plays a role in establishing the latent reservoir. These findings indicate that HIV-1 entry and integration do not trigger unique transcriptional responses when infection becomes latent.IMPORTANCELatent HIV-1 infection is established as early as the first viral exposure and remains the most important barrier in obtaining the cure for HIV-1 infection. Here, we used cap analysis of gene expression (CAGE) to compare the transcriptional landscape of latently infected cells with that of non-infected or productively infected cells. We found that latently infected cells and non-infected cells show quite similar transcriptional profiles. Our data suggest that T cells cannot recognize incoming viral components or the integrated HIV-1 genome when infection remains latent. These findings should guide future research into widening our approaches to identify and target latent HIV-1-infected cells.
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影响因子：
16.8
作者：
Pollizzi KN;Powell JD
通讯作者：
Powell JD
影响因子：
4.6
作者：
Cinti A;Le Sage V;Milev MP;Valiente-Echeverría F;Crossie C;Miron MJ;Panté N;Olivier M;Mouland AJ
通讯作者：
Mouland AJ
DOI：
10.1073/pnas.95.15.8869
发表时间：
1998-07-21
影响因子：
11.1
作者：
Chun, TW;Engel, D;Fauci, AS
通讯作者：
Fauci, AS
影响因子：
30.3
作者：
Baxter AE;Niessl J;Fromentin R;Richard J;Porichis F;Charlebois R;Massanella M;Brassard N;Alsahafi N;Delgado GG;Routy JP;Walker BD;Finzi A;Chomont N;Kaufmann DE
通讯作者：
Kaufmann DE
树突状细胞可有效清除 TCR 刺激之外的潜在 HIV-1，激活 PI3K-Akt-mTOR 通路
影响因子：
11.1
作者：
T. van Montfort;Renée M. Van der Sluis;G. Darcis;D. Beaty;K. Groen;A. Pasternak;G. Pollakis;Monique A. Vink;E. M. Westerhout;Mohamed Hamdi;M. Bakker;Boas van der Putten;S. Jurriaans;Jan H Prins;R. Jeeninga;Adri A. M. Thomas;D. Speijer;B. Berkhout
通讯作者：
B. Berkhout