Role of cell polarity regulators in HIV spreading

细胞极性调节剂在 HIV 传播中的作用

基本信息

批准号：
10013784
负责人：
HEINRICH GOTTLINGER
金额：
$ 53.28万
依托单位：
UNIV OF MASSACHUSETTS MED SCH WORCESTER
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-01-01 至 2024-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10013784
关键词：
Actins Antigen-Presenting Cells CD4 Positive T Lymphocytes CDC42 gene Cell Line Cell Polarity Cells Complex Cues Cytoskeleton Data Drug Targeting Eukaryotic Cell Exocytosis FMNL1 gene Family Filopodia Golgi Apparatus Guanosine Triphosphate Phosphohydrolases HIV HIV-1 Human Infection Investigation Knock-out Life Cycle Stages Mediating Membrane Microtubule-Organizing Center Microtubules Modeling Molecular Monomeric GTP-Binding Proteins PAK-1 kinase PARD6A gene Peripheral Blood Mononuclear Cell Phosphotransferases Polymerase Process Protein Kinase Proteins Public Health Reporting Role Secretory Vesicles Signal Pathway Site Surface T-Lymphocyte Vesicle Viral Virion Virus Yeasts cdc42 GTP-Binding Protein cytokine experimental study immunological synapse formation insight man polarized cell polymerization reconstitution response synaptogenesis trafficking transmission process virological synapse

项目摘要

ABSTRACT HIV-1-infected T cells can form stable conjugates with non-infected T cells in a process known as “virological synapse” formation. This process is reminiscent of the formation of an “immunological synapse”, during which CD4+ T cells rapidly polarize the actin cytoskeleton, the microtubule-organizing center (MTOC), and cytokine- containing vesicles towards antigen-presenting cells. The polarization of CD4+ T cells during immunological synapse formation depends on the RHO family GTPase CDC42, a molecular switch that has a key role in the establishment of polarity in eukaryotic cells. We have now observed that CDC42 is critical for the efficient spreading of HIV-1 in several T cell lines and in primary cells. However, our data also imply that CDC42 is dispensable for the completion of a single cycle of replication. Together, our observations implicate CDC42 in the cell-to-cell transmission of HIV-1. CDC42 stimulates the formation of membrane extensions, such as filopodia, through effectors that mediate the polarization of the actin cytoskeleton, and HIV-1 can exploit filopodial bridges to spread from cell to cell. Thus, our data let us to propose a working model in which CDC42 is crucial for the formation of intercellular extensions that facilitate the transfer of HIV-1 between CD4+ T cells. In support of this model, we have observed that CDC42 is required for the formation of HIV-1-induced membrane extensions by MOLT-3 cells. An alternative working model is that CDC42 is required for the polarized trafficking of HIV-1 virion components to the virologic synapse. We propose to directly examine the roles of CDC42 in HIV-1 cell-to-cell transmission and virological synapse formation, and to determine whether HIV-1 regulates the activity of CDC42. We also propose to examine the roles of CDC42 effectors that regulate localized actin assembly and polarized trafficking in HIV-1 spreading. Among these effectors are F-BAR proteins that connect to actin polymerization machinery, as does the F-BAR protein PACSIN2, which we have recently implicated in the cell-to-cell transmission of HIV-1. Notably, our preliminary results indicate that certain CDC42 effectors, including the CDC42-regulated actin polymerase FMNL1 and a putative CDC42 effector that controls polarized exocytosis, have crucial roles in HIV-1 replication. The proposed studies have the potential to yield fundamental new insights into the mechanism of an important but poorly understood mode of HIV-1 transmission. Of particular significance would be the identification of a kinase downstream of CDC42 as being critical for HIV-1 spreading, since protein kinases constitute one of the most important groups of drug targets.

抽象的 HIV-1感染的T细胞可以与未感染的T细胞形成稳定的结合物，这一过程被称为“病毒学” 这个过程让人想起“免疫突触”的形成。 CD4+ T 细胞快速极化肌动蛋白细胞骨架、微管组织中心 (MTOC) 和细胞因子含有针对抗原呈递细胞的囊泡的免疫学过程中 CD4+ T 细胞的极化。突触的形成依赖于 RHO 家族 GTPase CDC42，这是一种分子开关，在突触形成过程中发挥着关键作用。真核细胞极性的建立。我们现在观察到 CDC42 对于 HIV-1 在几种 T 细胞系中的有效传播至关重要然而，我们的数据也表明 CDC42 对于完成单个细胞是可有可无的。总之，我们的观察表明 CDC42 参与了 HIV-1 的细胞间传播。 CDC42 通过效应器刺激膜延伸的形成，例如丝状伪足介导肌动蛋白细胞骨架的极化，HIV-1 可以利用丝状伪足桥从细胞中传播因此，我们的数据让我们提出了一个工作模型，其中 CDC42 对于细胞的形成至关重要。促进 HIV-1 在 CD4+ T 细胞之间转移的细胞间延伸为了支持这一模型，我们。观察到 CDC42 是 MOLT-3 形成 HIV-1 诱导的膜延伸所必需的另一种工作模式是 CDC42 是 HIV-1 病毒颗粒极化运输所必需的。病毒突触的组成部分。我们建议直接检查 CDC42 在 HIV-1 细胞间传播和病毒学中的作用突触形成，并确定 HIV-1 是否调节 CDC42 的活性。研究 CDC42 效应器在 HIV-1 中调节局部肌动蛋白组装和极化运输的作用这些效应器中包括与肌动蛋白聚合机制相连的 F-BAR 蛋白。 F-BAR 蛋白 PACSIN2，我们最近发现它与 HIV-1 的细胞间传播有关。值得注意的是，我们的初步结果表明某些 CDC42 效应器，包括 CDC42 调节的肌动蛋白聚合酶 FMNL1 和控制极化胞吐作用的推定 CDC42 效应子在 HIV-1 复制。拟议的研究有可能对机制产生基本的新见解 HIV-1 传播的重要但知之甚少的模式尤其重要。鉴定出 CDC42 下游的激酶对于 HIV-1 传播至关重要，因为蛋白激酶是最重要的药物靶点之一。