BH3-only protein targeting by HHV-8

HHV-8 仅靶向 BH3 蛋白

• 批准号: 9193611
• 负责人: John Nicholas
• 金额: $ 40.5万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-01-01 至 2018-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9193611
• 关键词: Agonist; Antiviral Agents; Apoptosis; Apoptotic; B lymphoid malignancy; BH3 Domain; BIM Bcl-2-binding protein; Binding; Biological; Biological Assay; CCR8 gene; Cell Survival; Cells; Coupled; Custom; Defense Mechanisms; Detection; Development; Disease; Endothelial Cells; Etiology; Future; G Protein-Coupled Receptor Signaling; G-Protein-Coupled Receptors; G-substrate; GTP-Binding Proteins; Genetic Transcription; HIV; Homologous Gene; Human Herpesvirus 8; Immune; In Vitro; Individual; Infection; Kaposi Sarcoma; Laboratories; Lead; Link; Lytic; Lytic Phase; Mediating; Mediator of activation protein; Methods; Modeling; Multicentric Angiofollicular Lymphoid Hyperplasia; Pathology; Pathway interactions; Peptides; Phage Display; Pharmaceutical Preparations; Pharmacology; Phosphorylation; Physiologic pulse; Post-Translational Regulation; Process; Production; Protein Binding Domain; Proteins; Public Health; Reagent; Receptor Signaling; Regulation; Reporting; Research; Role; Signal Pathway; Signal Transduction; Stress; System; Testing; Therapeutic; Transcriptional Regulation; Ubiquitination; Variant; Viral; Viral Load result; Viral Physiology; Viral Proteins; Virus; Virus Diseases; Virus Inhibitors; Virus Receptors; Virus Replication; chemokine; chemokine receptor; co-infection; disorder prevention; experimental study; gene product; genetic resistance; inhibitor/antagonist; lytic replication; novel; prevent; primary effusion lymphoma; protein function; public health relevance; small hairpin RNA; small molecule inhibitor; viral interferon regulatory factor; viral interferon regulatory factor-1

DESCRIPTION (provided by applicant): Human herpesvirus 8 (HHV-8) is associated with endothelial Kaposi's sarcoma (KS) and B cell malignancies primary effusion lymphoma and multicentric Castleman's disease, all of which occur predominantly in the context of HIV coinfection. Both latent and lytic gene products are believed to contribute to these pathologies, and productive replication and increased viral loads are associated with KS. Understanding the processes required for successful lytic replication will provide opportunities for targeting essential viral functions to block virus replication and thereby treat or prevent HHV-8 associated disease. Key to replication by HHV-8 and other viruses are viral mechanisms that limit stress-induced apoptotic signaling that serves as an innate defense mechanism against virus infection. HHV-8 encodes several proteins that inhibit such pro-apoptotic pathways. These proteins include viral interferon regulatory factor-1 (vIRF-1), viral G protein coupled receptor (vGPCR), and viral chemokines vCCL-1 and vCCL-2, all of which have been demonstrated to both inhibit apoptosis and be required for efficient virus replication. This laboratory has identified inhibitor interactions of vIRF-1 with replication-induced pro-apoptotic BH3-only proteins (BOPs) Bim and Bid and revealed the critical importance of controlling these proteins for efficient virus replicaton to occur. Suppression of Bim or Bid expression via transduced shRNAs leads to very significant increases in virus replication, and disruption of vIRF-1:BOP interactions inhibits virus production and promotes apoptosis in lytically infected cells. In addition to Bim and Bid, vIRF-1 targets other BOPs (Bik, Bmf, Hrk, Noxa), via their functional BH3 domains, indicating the biological importance of these BOPs also and the need to inhibit their activities during lytic replication. Furthermore, the viral chemokine receptor (vGPCR) and CCR8 agonists vCCL-1 and vCCL-2 activate signaling pathways that lead to suppression of Bim expression. Thus, chemokine receptor signaling is likely to enhance HHV-8 productive replication in part via BOP regulation. This application proposes to: (1) elucidate the mechanisms of vGPCR and vCCL suppression of Bim, an identified major inhibitor of HHV-8 replication; (2) identify peptide and pharmacological inhibitors of vIRF-1:BOP interactions to specifically inhibit this mechanism of BOP regulation; (3) determine the antiviral activities of lytic cycle-induced BOPs and the individual and combined contributions, via BOP control, of vCCLs, vGPCR, and vIRF-1 to HHV- 8 productive replication. The project comprises a focused analysis of BOP function and targeting by vCCLs, vGPCR and vIRF-1 during HHV-8 lytic replication and assessment of inhibitory reagents and methods that could potentially be developed for antiviral purposes.

描述（由申请人提供）：人疱疹病毒8（HHV-8）与内皮kaposi的肉瘤（KS）和B细胞恶性肿瘤有关原发性积液淋巴瘤和多中性骑士氏病，所有这些都主要出现在HIV共同体的背景下。据信潜在基因和裂解基因产物都会有助于这些病理，而生产性复制和增加的病毒载量与KS相关。了解成功的裂解复制所需的过程将为靶向基本病毒功能以阻止病毒复制，从而为治疗或预防HHV-8相关疾病提供机会。 HHV-8和其他病毒复制的关键是病毒机制，这些机制限制了应激诱导的凋亡信号传导，它是针对病毒感染的先天防御机制。 HHV-8编码几种抑制这种促凋亡途径的蛋白质。这些蛋白质包括病毒干扰素调节因子-1（VIRF-1），病毒G蛋白偶联受体（VGPCR）和病毒趋化因子VCCL-1和VCCL-2，所有这些都已抑制凋亡，并且需要有效地凋亡。病毒复制。该实验室已经确定了VIRF-1与复制诱导的促凋亡BH3蛋白（BOPS）BIM和BIM的抑制剂相互作用，并揭示了控制这些蛋白质以进行有效的病毒复制品发生的至关重要性。通过转导的SHRNA抑制BIM或投标表达，导致病毒复制的显着增加，而破坏了VIRF-1：BOP相互作用抑制了病毒的产生 并促进溶血感染细胞的凋亡。除了BIM和BID外，VIRF-1还通过其功能性BH3域靶向其他BOP（BIK，BMF，HRK，NOXA），这表明这些BOPS的生物学重要性也需要抑制其在裂解复制过程中的活性。此外，病毒趋化因子受体（VGPCR）和CCR8激动剂VCCL-1和VCCL-2激活了导致BIM表达抑制的信号通路。因此，趋化因子受体信号传导可能会通过BOP调节部分增强HHV-8的生产复制。该应用建议：（1）阐明了BIM的VGPCR和VCCL抑制的机制，BIM是HHV-8复制的主要抑制剂； （2）确定VIRF-1的肽和药理抑制剂：BOP相互作用以特异性抑制这种BOP调节机制； （3） 通过BOP控制，VCCL，VGPCR和VIRF-1对HHV-8的生产性复制，确定裂解循环诱导的BOP的抗病毒活性以及个人和组合的贡献。该项目包括对HHV-8裂解复制过程中VCCL，VGPCR和VIRF-1靶向的重点分析，并评估抑制性试剂和方法可能是出于抗病毒目的而开发的。