iTRAQ-based proteomic study on monocyte cell model discovered an association of LAMP2 downregulation with HIV-1 latency.

IF 2.1 3区生物学 Q3 BIOCHEMICAL RESEARCH METHODS Proteome Science Pub Date : 2024-05-15 DOI:10.1186/s12953-024-00230-3

Lin Yin, Qimin Wang, Siyuan Liu, Jun Chen, Yujiao Zhang, Lingqing Lu, Hongzhou Lu, Zhigang Song, Lijun Zhang

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Abstract

Background: Patients with immunodeficiency virus-1 (HIV-1) infection are challenging to be cured completely due to the existence of HIV-1 latency reservoirs. However, the knowledge of the mechanisms and biomarkers associated with HIV-1 latency is limited. Therefore, identifying proteins related to HIV-1 latency could provide new insights into the underlying mechanisms of HIV-1 latency, and ultimately contribute to the eradication of HIV reservoirs.

Methods: An Isobaric Tags for Relative and Absolute Quantification (iTRAQ)-labeled subcellular proteomic study was performed on an HIV-1 latently infected cell model (U1, a HIV-1-integrated U937 cell line) and its control (U937). Differentially expressed proteins (DEPs) were analyzed using STRING-DB. Selected DEPs were further evaluated by western blotting and multiple reaction monitoring technology in both cell model and patient-derived cluster of differentiation 4 (CD4)⁺ T cells. Finally, we investigated the relationship between a specific DEP lysosome-associated membrane glycoprotein 2 (LAMP2) and HIV-1 reactivation by panobinostat or lysosome regulation by a lysosomotropic agent hydroxychloroquine in U1 and U937 cells.

Results: In total, 110 DEPs were identified in U1 cells comparing to U937 control cells. Bioinformatics analysis suggested associations of the altered proteins with the immune response and endosomal/lysosomal pathway. LAMP2, leukocyte surface antigen CD47, CD55, and ITGA6 were downregulated in HIV-1 latent cells. Downregulated LAMP2 was further confirmed in resting CD4⁺ T cells from patients with latent HIV-1 infection. Furthermore, both HIV-1 reactivation by panobinostat and stimulation with hydroxychloroquine upregulated LAMP2 expression.

Conclusions: Our results indicated the involvement of the endosomal/lysosomal pathway in HIV-1 latency in macrophage cell model. The down-modulation of LAMP2 was associated with HIV latency, and the restoration of LAMP2 expression accompanied the transition of viral latency to active infection. This study provides new insights into the mechanism of HIV-1 latency and potential strategies for eradicating HIV-1 reservoirs by targeting LAMP2 expression.

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基于 iTRAQ 的单核细胞模型蛋白质组学研究发现，LAMP2 的下调与 HIV-1 潜伏期有关。

背景：由于 HIV-1 潜伏库的存在，免疫缺陷病毒-1（HIV-1）感染患者很难被完全治愈。然而，人们对与 HIV-1 潜伏期相关的机制和生物标志物的了解十分有限。因此，鉴定与HIV-1潜伏期相关的蛋白质可以为了解HIV-1潜伏期的内在机制提供新的视角，并最终为根除HIV病毒库做出贡献：方法：对HIV-1潜伏感染细胞模型（U1，一种HIV-1整合的U937细胞系）及其对照（U937）进行了等位基因标记的亚细胞蛋白质组学研究。使用 STRING-DB 分析了差异表达蛋白（DEPs）。在细胞模型和患者来源的分化群 4 (CD4)+ T 细胞中，通过 Western 印迹和多反应监测技术进一步评估了所选的 DEPs。最后，我们在 U1 和 U937 细胞中研究了特定 DEP 溶酶体相关膜糖蛋白 2（LAMP2）与泛比诺司他激活 HIV-1 或羟氯喹溶酶体调节剂调节溶酶体之间的关系：结果：与 U937 对照细胞相比，U1 细胞中共鉴定出 110 个 DEPs。生物信息学分析表明，改变的蛋白质与免疫反应和内体/溶酶体途径有关。LAMP2、白细胞表面抗原CD47、CD55和ITGA6在HIV-1潜伏细胞中下调。下调的 LAMP2 在潜伏 HIV-1 感染者的静息 CD4+ T 细胞中得到了进一步证实。此外，帕诺比诺司他（panobinostat）和羟氯喹（hydroxychloroquine）刺激的HIV-1再激活都会上调LAMP2的表达：我们的研究结果表明，在巨噬细胞模型中，内体/溶酶体途径参与了HIV-1潜伏。结论：我们的研究结果表明，在巨噬细胞模型中，内体/溶酶体途径参与了HIV-1的潜伏，LAMP2的下调与HIV潜伏有关，而LAMP2表达的恢复则伴随着病毒潜伏向活动感染的转变。这项研究提供了关于HIV-1潜伏机制的新见解，以及通过靶向LAMP2表达消除HIV-1储库的潜在策略。

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来源期刊

Proteome Science 生物-生化研究方法

CiteScore

2.90

自引率

0.00%

发文量

审稿时长

4.5 months

期刊介绍： Proteome Science is an open access journal publishing research in the area of systems studies. Proteome Science considers manuscripts based on all aspects of functional and structural proteomics, genomics, metabolomics, systems analysis and metabiome analysis. It encourages the submissions of studies that use large-scale or systems analysis of biomolecules in a cellular, organismal and/or environmental context. Studies that describe novel biological or clinical insights as well as methods-focused studies that describe novel methods for the large-scale study of any and all biomolecules in cells and tissues, such as mass spectrometry, protein and nucleic acid microarrays, genomics, next-generation sequencing and computational algorithms and methods are all within the scope of Proteome Science, as are electron topography, structural methods, proteogenomics, chemical proteomics, stem cell proteomics, organelle proteomics, plant and microbial proteomics. In spite of its name, Proteome Science considers all aspects of large-scale and systems studies because ultimately any mechanism that results in genomic and metabolomic changes will affect or be affected by the proteome. To reflect this intrinsic relationship of biological systems, Proteome Science will consider all such articles.