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: 9359351
• 负责人: JOSEPH S PAGANO
• 金额: $ 31.7万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/9359351
• 关键词: Affect; B-Cell Lymphomas; B-Lymphocytes; Binding; Biochemical; Biogenesis; Carcinoma; Cell Culture Techniques; Cell physiology; Cells; Cellular Structures; Communication; Complex; Computer Simulation; Controlled Study; Data; Deubiquitinating Enzyme; Deubiquitination; Development; Disease; Ensure; Epithelial; Epithelial Cells; Epstein-Barr Virus latency; Epstein-Barr virus LMP-1 protein; FGF2 gene; 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 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; Tumorigenicity; Ubiquitin; Ubiquitin C; Ubiquitin family; Vascular Endothelial Growth Factors; Vesicle; Viral; 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; response; small molecule inhibitor; trafficking; transforming virus; 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