Analysis and Therapeutic Targeting of the Linear-Ubiquitination Pathway in Hodgkin Lymphoma

霍奇金淋巴瘤线性泛素化途径的分析和治疗靶向

• 批准号: 10210535
• 负责人: Yibin Yang
• 金额: $ 42.78万
• 依托单位: RESEARCH INST OF FOX CHASE CAN CTR
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-06-01 至 2026-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10210535
• 关键词: Address; Antigens; Apoptosis; B cell differentiation; B-Lymphocytes; Biological Assay; Biology; C-terminal; CRISPR library; CRISPR screen; Cell Survival; Cell surface; Cells; Clinical; Coculture Techniques; Complex; Development; Diagnosis; Disease; Enzymes; Gene Expression Profiling; Genes; Growth; Hodgkin Disease; Human; Human Herpesvirus 4; Immune; Immune checkpoint inhibitor; Immune response; Immune system; Immunotherapy; In Vitro; In complete remission; Inflammatory; Interleukin-13; Intervention; Knowledge; Lead; Libraries; Lymphoma; Malignant - descriptor; Malignant lymphoid neoplasm; Mediating; Minority Groups; Molecular; Mus; Mutation; Outcome; Pathogenesis; Pathologic; Pathway interactions; Patients; Peripheral Blood Mononuclear Cell; Phenotype; Process; Production; Proteins; Proteomics; Public Health; Recurrence; Reed-Sternberg Cells; Refractory; Refractory Disease; Relapse; Role; STAT6 gene; Sampling; Signal Transduction; Survival Rate; System; T-Cell Proliferation; TNFRSF8 gene; Testing; Therapeutic; Translating; Tumor Escape; Ubiquitin; Ubiquitination; Xenograft Model; Zinc Fingers; base; cell growth; cytokine; efficacy evaluation; humanized mouse; immune checkpoint; immune clearance; improved; in vivo; in vivo evaluation; inhibitor/antagonist; insight; loss of function; molecular pathology; mutant; mutational status; neoplastic cell; novel; novel strategies; novel therapeutic intervention; prevent; programmed cell death ligand 1; programs; rare cancer; response; screening; small molecular inhibitor; targeted treatment; therapeutic development; therapeutic target; therapeutically effective; transcriptome sequencing; treatment strategy; tumor; tumor growth; tumor-immune system interactions; young adult

PROJECT SUMMARY Hodgkin lymphoma (HL) is the most common (6,000 to 7,000 new cases per year) lymphoma subtype in young adulthood. Although there has been great progress during the last few decades, the survival rate for patients diagnosed at an advanced stage or with relapsed/refractory disease remains low. The current understanding of the biology of the disease has been translated into the development and approval of therapeutic agents that target HL-specific antigens or immune checkpoint pathways. However, the number of patients in complete remission has been low, and relapse frequently develops, leading to poor outcomes. Thus, a clearer understanding of the molecular pathology of HL is necessary to develop new treatment strategies. HL is characterized by a minority population of malignant Hodgkin and Reed/Sternberg (HRS) cells in a background of dense inflammatory cells. HRS cells have lost their B cell phenotype, however, and escaped from BCR- mediated apoptosis and immune elimination. Therefore, two major questions remain: (a), how do HRS cells escape the control of the immune system; (b), how do they survive despite the absence of BCR expression? To address these gaps in knowledge, we applied an unbiased high throughput CRISPR screening, and identified an essential role for the linear ubiquitin chain assembly complex (LUBAC) in HL pathogenesis. Our preliminary studies demonstrated that LUBAC activity promotes HRS cell survival and immune escape, which significantly overlaps with mutation status of the most recurrent genetically altered gene in HL, A20. Clinically, LUBAC activity is consistently elevated in most primary HL cases, and this is correlated with low A20 expression. Moreover, using RNA-seq analysis, we identified a set of LUBAC-regulated genes in HL that overlapped significantly with signatures reflecting NF-κB and JAK-STAT activities, as well as TH2 cytokines and cell surface immunosuppressive molecules. Unexpectedly, our BioID proteomic screening reveals a CD30-LUBAC complex in HL, suggesting the role of LUBAC-A20 axis in the CD30 mediated NF-κB signaling, TH2 cytokine production, and STAT6 activation. Finally, a highly specific LUBAC small molecular inhibitor shows promising activity against HL in vitro and in a mouse xenograft model. Altogether, these findings provide strong support for our hypothesis that the LUBAC-A20 axis regulates HL pathogenesis, and that targeting LUBAC could be a novel therapeutic strategy in this disease. In this study, we will: 1) investigate the mechanistic basis by which the LUBAC-A20 axis supports HRS survival and proliferation; 2) evaluate how linear-ubiquitin-dependent signaling regulates the molecular circuitry that drives tumor immune escape of HL; and 3) exploit the therapeutic potential of targeting LUBAC to provide novel intervention strategies for both targeted and immune therapies in HL. These studies promise to reveal critical insights into the molecular circuitry that drives this lymphoid cancer, as well as provide unique opportunities for the development of novel strategies for both targeted and immune therapies to treat HL.

项目摘要 霍奇金淋巴瘤（HL）是年轻人中最常见的（每年6，000至7，000例新发病例）淋巴瘤亚型。 成年虽然在过去的几十年里有了很大的进步，但患者的生存率 诊断为晚期或复发/难治性疾病的患者仍然很少。目前了解的 该疾病的生物学已经转化为治疗剂的开发和批准， 靶向HL特异性抗原或免疫检查点途径。然而，完成的患者人数 缓解率很低，经常复发，导致不良结果。因此，更清晰 对HL的分子病理学的理解对于开发新的治疗策略是必要的。HL是 特征在于背景中的恶性霍奇金和里德/斯滕贝格（HRS）细胞的少数群体 密集的炎症细胞。然而，HRS细胞已经失去了它们的B细胞表型，并从BCR-1中逃脱。 介导的细胞凋亡和免疫消除。因此，两个主要问题仍然存在：（a），HRS细胞如何 逃脱免疫系统的控制;（B），尽管缺乏BCR表达，它们如何存活？到 为了解决这些知识差距，我们应用了无偏见的高通量CRISPR筛选，并确定了 线性泛素链组装复合物（LUBAC）在HL发病机制中的重要作用。我们的初步 研究表明，LUBAC活性促进HRS细胞存活和免疫逃逸， 与HL A20中最常见的遗传改变基因的突变状态重叠。临床上，LUBAC活性 在大多数原发性HL病例中持续升高，这与低A20表达相关。此外，委员会认为， 使用RNA-seq分析，我们在HL中鉴定了一组LUBAC调控的基因，这些基因与 反映NF-κB和JAK-STAT活性以及TH 2细胞因子和细胞表面的特征 免疫抑制分子出乎意料的是，我们的BioID蛋白质组学筛选揭示了CD 30-LUBAC复合物 在HL中，提示LUBAC-A20轴在CD 30介导的NF-κB信号传导，TH 2细胞因子产生， STAT 6激活。最后，一种高度特异性的LUBAC小分子抑制剂显示出有希望的活性， HL在体外和小鼠异种移植模型中。总之，这些发现为我们的假设提供了强有力的支持 LUBAC-A20轴调节HL发病机制，靶向LUBAC可能是一种新的治疗方法， 在这种疾病的战略。在这项研究中，我们将：1）调查的机制基础，LUBAC-A20 轴支持HRS的生存和增殖; 2）评估线性泛素依赖性信号转导如何调节HRS的生存和增殖。 驱动HL的肿瘤免疫逃逸的分子电路;和3）利用靶向的治疗潜力 LUBAC为HL的靶向和免疫治疗提供新的干预策略。这些研究 有望揭示驱动这种淋巴癌的分子回路的关键见解，并提供 为HL的靶向和免疫治疗开发新策略提供了独特的机会。