Project 2 - Membrane trafficking in EBV malignancies: Implication of deubiquitinase UCH-L1

项目 2 - EBV 恶性肿瘤中的膜运输：去泛素酶 UCH-L1 的影响

• 批准号: 9073484
• 负责人: JOSEPH S PAGANO
• 金额: $ 30.88万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-05-01 至 2021-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9073484
• 关键词: Affect; B-Cell Lymphomas; B-Lymphocytes; Binding; Biochemical; Biogenesis; Carcinoma; Cell Communication; Cell Culture Techniques; Cell physiology; Cells; Cellular Structures; Complex; Computer Simulation; Controlled Study; Data; Deubiquitinating Enzyme; Deubiquitination; Development; Disease; Ensure; Epithelial; Epithelial Cells; Epstein-Barr virus LMP-1 protein; FGF2 gene; Family; Genes; Human; Human Herpesvirus 4; Human Herpesvirus 8; Human Papillomavirus; Hydrolase; Infection; Intracellular Membranes; Link; Lymphoid Cell; Lymphoma; Lymphomagenesis; MMP9 gene; Malignant Neoplasms; Malignant lymphoid neoplasm; Mediating; Membrane; Membrane Microdomains; Membrane Protein Traffic; Membrane Proteins; Mesenchymal; Metastatic Carcinoma; Molecular; N-Cadherin; Nanotubes; Neoplasm Metastasis; Oncogenes; Oncogenic Viruses; Outcome; Pathogenesis; Pathway interactions; Patients; Physiological; Play; Preclinical Drug Evaluation; Primary Neoplasm; Process; Production; Property; Proteins; Role; Sorting - Cell Movement; Stromal Cells; Stromal Neoplasm; Structure; System; Testing; Tissues; Tube; Ubiquitin; Ubiquitin C; Vascular Endothelial Growth Factors; Vesicle; Viral; abstracting; base; cancer cell; cell transformation; enzyme activity; exosome; extracellular; humanized mouse; in vivo; inhibitor/antagonist; interest; late endosome; membrane biogenesis; microvesicles; mouse model; novel; prevent; reactivation from latency; response; small molecule inhibitor; trafficking; tumor; tumor microenvironment; tumor progression; tumorigenesis; tumorigenic; ubiquitin C-terminal hydrolase

Project 2 Project Summary Abstract The implication of extracellular membrane vesicles such as exosomes, and membrane protrusions such as tunneling nanotubes, in intercellular trafficking networks opens a new perspective in understanding tumor development and progression. The ubiquitin system is one of the central regulators of biogenesis and function of exosomes and nanotubes, and infection with tumor viruses including Epstein-Barr Virus (EBV) results in deregulation of cellular functions by manipulation of this system during cell transformation. Studies in recent years clearly demonstrate that the spectrum of potential functions of the small evolutionarily conserved family of Ubiquitin C-terminal Hydrolases (UCHs) is much wider than was suspected. Among them, UCH-l1 is of special interest: recent studies including ours demonstrate that this unique deubiquitinase is closely involved not only in cell transformation and in primary tumor formation, but is a main regulator of cancer progression as well. Based directly on our previous studies and substantial preliminary data, we hypothesize that UCH-L1 is a major regulator of ubiquitin-dependent processes of intra- and inter-cellular trafficking in EBV-positive cancers. In Aim I, we will analyze how distinct biochemical functions of UCH-L1 are required for exosome biogenesis and sorting, and how the EBV major oncogene, Latent Membrane Protein 1 (LMP1), is involved in these processes. In Aim II, based on our recent discovery that N-cadherin is highly expressed in EBV-driven B-cell lymphomas, and co-localizes with UCH-L1 in these cells, we will explore the role of N-cadherin-based complexes in intercellular trafficking of pro-metastatic factors produced by EBV-positive cancer cells. The results in this Aim will clarify how tunneling nanotubes and exosomes produced by EBV-transformed cells change the tumor microenvironment by transferring pro-invasive factors to tumor stromal tissues. In Aim III we will investigate whether inhibition of UCH-L1 activity with specific small-molecule inhibitors that have anti- tumorigenic effects in cell culture will also be active in vivo against EBV B-cell lymphogenesis in humanized mice. We will determine whether UCH-L1 DUB activity is required for EBV-induced immortalization of normal human B-cells, and whether inhibition of UCH-L1 affects expression and function of EBV genes in latently infected cells as well as during viral reactivation. EBV is tightly linked to several highly invasive malignancies of lymphoid and epithelial origin; treatment of patients with these malignancies poses unique challenges, and outcomes remain poor. Since recent study has demonstrated a profound anti-metastatic effect of such UCH-L1 inhibitor in a mouse model of invasive carcinoma, specific inhibitors of UCH-L1 enzymatic activity may offer an adjunct to existing therapies.

项目2 项目摘要摘要 细胞外膜囊泡，如外切体，以及膜突起，如 在细胞间运输网络中隧道传输纳米管为理解肿瘤打开了一个新的视角 发展和进步。泛素系统是生物发生和功能的中央调节系统之一 外切体和纳米管，以及感染包括EB病毒(EBV)在内的肿瘤病毒导致 在细胞转化过程中通过操纵这一系统来解除对细胞功能的调节。近年来的研究 年清楚地表明，进化保守的小家庭的势函数谱 泛素C末端水解酶(UCH)的范围比人们想象的要广泛得多。其中，UCH-L1是 特别兴趣：最近的研究包括我们的研究表明，这种独特的脱泛素酶密切相关 不仅在细胞转化和原发肿瘤形成中，而且是癌症进展的主要调节因子，如 井。直接基于我们之前的研究和大量的初步数据，我们假设UCH-L1是一种 EBV阳性癌症细胞内和细胞间转运的泛素依赖过程的主要调节因子。 在目标I中，我们将分析UCH-L1的不同生化功能对于外切体的生物发生是如何需要的 以及EBV的主要癌基因潜伏膜蛋白1(LMP1)是如何参与这些过程的 流程。在AIM II中，基于我们最近发现N-钙粘蛋白在EBV驱动的B细胞中高表达 淋巴瘤，并与UCH-L1在这些细胞中共定位，我们将探索基于N-钙粘附素的作用 EBV阳性癌细胞产生的促转移因子的细胞间转运复合体。这个 这一目标的结果将阐明隧道传输纳米管和外切体是如何由EBV转化的细胞产生的 通过将促侵袭因子转移到肿瘤间质组织来改变肿瘤微环境。在AIM III中，我们 将调查是否使用具有抗-L1活性的特定小分子抑制剂来抑制UCH-L1活性 细胞培养中的致瘤作用也将在体内活跃起来，以对抗人源化的EBV B细胞淋巴生成 老鼠。我们将确定UCH-L1 DUB活性是否为EBV诱导的正常人永生化所必需的 人类B细胞，以及抑制UCH-L1是否潜伏影响EBV基因的表达和功能 在感染细胞和病毒重新激活期间也是如此。EBV与几种高度侵袭性的恶性肿瘤密切相关 淋巴和上皮起源；这些恶性肿瘤患者的治疗构成了独特的挑战， 结果仍然不佳。由于最近的研究表明这种UCH-L1具有深刻的抗转移作用 在小鼠浸润性癌模型中，UCH-L1酶活性的特异性抑制剂可能提供一种 作为现有疗法的补充。