A radiation-induced cellular stress activates HIV and induces killing of infected cells

辐射引起的细胞应激会激活艾滋病毒并诱导杀死受感染的细胞

• 批准号: 9326140
• 负责人: Fatah Kashanchi
• 金额: $ 22.8万
• 依托单位: GEORGE MASON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-08-15 至 2019-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9326140
• 关键词: Agonist; Anti-Retroviral Agents; Apoptosis; Autophagocytosis; Binding; CASP1 gene; CD4 Positive T Lymphocytes; Calpain; Cell Cycle; Cell Death; Cell Line; Cell Survival; Cells; Cellular Stress; Cessation of life; Chronic; Cultured Cells; Cyclins; DNA Damage; DNA Polymerase II; Data; Dose; Epigenetic Process; Genetic Transcription; Genome; Goals; HDAC1 gene; HIV; HIV-1; Highly Active Antiretroviral Therapy; Human; Human immunodeficiency virus test; Immune response; Immune system; Individual; Induction of Apoptosis; Interleukin-7; Methods; Mus; Nuclear; Organ; Outcome; Patients; Pharmaceutical Preparations; Phosphorylation; Plasma; Population; Positive Transcriptional Elongation Factor B; Production; Proteasome Inhibitor; Publishing; RNA; RNA Polymerase II; Radiation; Rest; Reverse Transcriptase Polymerase Chain Reaction; Roentgen Rays; S Phase; Scheme; Shock; Signal Transduction; Specificity; Stress; T-Lymphocyte; TP53 gene; Testing; Therapeutic; Tissues; Transcriptional Activation; Transferase; Viral; Viral Proteins; Viral reservoir; Virus; Virus Replication; Western Blotting; bryostatin; cancer therapy; chromatin immunoprecipitation; cytotoxic; humanized mouse; in vivo; inhibitor/antagonist; irradiation; killings; macrophage; promoter; repaired; response; transcription factor; viral RNA

HIV-1 can be preserved in the long-lived resting CD4+ T cells which form a viral reservoir in infected individuals. This reservoir may persist for many years even though the patients are usually treated with highly active antiretroviral therapy (HAART), and viral population can be recovered once HAART is stopped. Thus, the selective activation of the latently HIV-infected cells resulting in the replication of proviral genome is critical to make infected cells recognized by immune system. Our long term goal is to find the method of reactivation of HIV from latency and killing of infected cells that does not kill or destroy non-infected quiescent cells and subsequently target and eradicate the persistent HIV-1 reservoirs. We have preliminary data showing that a well-characterized stress signal, such as irradiation (IR), activated inappropriate entry of the HIV infected T cells to S phase of the cell cycle and increased viral transcription and eventual apoptosis of infected cells. Importantly, the parental uninfected cells did not demonstrate this response to the same irradiation dose. Our recently published data elucidated this phenomenon and indicated (1) increase of HIV-1 transcription via epigenetic mechanisms after the IR-induced DNA damage, as evidenced by the presence of RNA polymerase II and reduction of HDAC1 and methyl transferase SUV39H1 on the HIV-1 promoter; (2) elevated level of intracellular HIV-1 RNA and increased expression of viral proteins in the IR-treated HIV-1 infected quiescent CD4+ T cells; (3) enhancement of transcription activation in latently HIV-1infected macrophages treated with PKC agonist bryostatin 1 after IR; (4) higher death of irradiated HIV-1 chronically-infected cells via increased phosphorylation of Ser46 in p53 that is responsible for apoptosis induction. Finally, (5) exposure of HIV-1 infected humanized mice with undetectable viral RNA level to IR resulted in a significant increase of HIV-1 RNA in plasma and certain tissue viral reservoirs. We hypothesize, that the cellular stress induced by low IR doses reactivates HIV-1 transcription from latently infected cells resulting in nuclear accumulation of Tat, activation of HIV-1 genome expression and enhanced apoptosis of infected, but not uninfected, cells. Our two aims include; A) To identify the mechanism of HIV-1 reactivation in response to therapeutic X-ray doses in latently infected T cells and to assess IR effect on the Tat activated HIV-1 transcription; and B) To decipher mechanism of enhanced apoptosis of HIV-1 infected cells in response to IR doses. The expected outcomes of these proposed studies include elucidation of the mechanisms that determine HIV-1 reactivation and apoptosis in latently infected cells and tissues in response to the X ray IR. We also expect to determine optimal IR dose that does not destroy uninfected cells, but induces HIV-1 reactivation and apoptosis of latently infected cells.

HIV-1可以保存在长寿命的静息CD4+ T细胞中，该细胞在感染中形成病毒储量 个人。即使患者通常接受治疗，该水库也可能持续多年 使用高活性的抗逆转录病毒疗法（HAART），并且可以回收病毒种群 哈特停止了。因此，潜在的HIV感染细胞的选择性激活导致 病毒基因组的复制对于使被免疫系统识别的感染细胞至关重要。我们的 长期目标是从潜伏期和杀死感染细胞中找到HIV重新激活的方法 这不会杀死或破坏未感染的静态细胞，然后靶向并消除 持续的HIV-1水库。我们有初步数据表明，特征良好的压力 信号，例如照射（IR），激活了HIV感染的T细胞的不适当进入 细胞周期并增加了感染细胞的病毒转录和最终凋亡。重要的是， 父母未感染的细胞未证明对相同辐照剂量的这种反应。我们的 最近发布的数据阐明了这一现象，并指出了（1）HIV-1的增加 IR诱导的DNA损伤后通过表观遗传机制的转录，这证明了 RNA聚合酶II的存在以及HDAC1和甲基转移酶SUV39H1的降低 HIV-1启动子； （2）细胞内HIV-1 RNA的水平升高，病毒的表达增加 IR处理的HIV-1感染静态CD4+ T细胞中的蛋白质； （3）增强转录 IR后用PKC激动剂Bryostatin 1处理的潜在HIV-1感染的HIV-1感染的巨噬细胞激活； （4） 通过增加的Ser46磷酸化在慢性的HIV-1慢性感染细胞中死亡 负责凋亡诱导的p53。最后，（5）暴露于HIV-1感染人性化的 IR无法检测到的病毒RNA水平的小鼠导致HIV-1 RNA在 血浆和某些组织病毒储层。我们假设，低IR诱导的细胞应激 剂量使来自潜在感染细胞的HIV-1转录重新激活，导致核积累 TAT，HIV-1基因组表达的激活和受感染但未感染的凋亡增强， 细胞。我们的两个目标包括： a）确定响应于 治疗性X射线剂量在潜在感染的T细胞中，并评估IR对TAT激活HIV-1的影响 转录； b）为破译HIV-1感染细胞凋亡增强的机理 对IR剂量的反应。这些提出的研究的预期结果包括阐明 确定HIV-1重新激活和凋亡的机制 对X射线IR的响应。我们还期望确定不会破坏的最佳红外剂量 未感染的细胞，但诱导了潜在感染细胞的HIV-1重新激活和凋亡。