Trib1 in NF-kappaB Signaling: Insights into MALT1 Regulation and Leukemia

NF-kappaB 信号传导中的 Trib1：深入了解 MALT1 调节和白血病

• 批准号: 8783330
• 负责人: Kelly Rome
• 金额: $ 4.27万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-01 至 2017-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8783330
• 关键词: A20 protein; Acute Myelocytic Leukemia; Apoptosis; Binding; Biological Assay; Bone Marrow; Cell Proliferation; Cell Survival; Cell physiology; Cells; Co-Immunoprecipitations; Collaborations; Data; Dependence; Development; Developmental Process; Diagnosis; Disease; Drosophila genus; Genetic; Hematologic Neoplasms; Hematopoietic; Homologous Gene; Human; Knowledge; Leukemic Cell; Link; Lymphocyte; Maintenance; Maps; Mediating; Mediator of activation protein; Monoubiquitination; Mus; Myelogenous; NF-kappa B; Oncogenes; Oncogenic; Pathogenesis; Pathway interactions; Patients; Peptide Hydrolases; Phosphorylation; Polyubiquitination; Proteins; Proteomics; Publishing; Regulation; Reporter; Role; Sampling; Signal Transduction; Small Interfering RNA; Specificity; Structural Biologist; Structure; Therapeutic; Therapeutic Intervention; Ubiquitin; Ubiquitination; Universities; Wild Type Mouse; Work; base; cell motility; improved; inhibitor/antagonist; insight; leukemia; mouse model; mutant; novel; outcome forecast; overexpression; progenitor; protein function; public health relevance; reconstitution; research study; retroviral transduction; scaffold; small molecule; ubiquitin-protein ligase

DESCRIPTION (provided by applicant): Through the proposed work, I will characterize the role of Tribbles homolog 1 (Trib1) in NF?B signaling, and identify the implications of this role in the pathogenesis of Acute Myeloid Leukemia (AML). We identified MALT1 as a Trib1interacting protein in a screen to identify novel Trib binding partners. Trib1 overexpression in the Jurkat Tcell line enhanced NF?B activity in a NF?Bresponsive reporter assay. Given the function of MALT1 in mediating NF?B signaling, I hypothesize that Trib1 enhances NF?B activation by regulating MALT1 activity. Published work from my lab identified Trib1 as an oncogene in AML. Retroviral transduction of Trib1 in bone marrow progenitors induces AML in reconstituted mice. Furthermore, Trib1 is highly expressed in multiple human AML patient subsets. I hypothesize that Trib1 promotes AML pathogenesis by enhancing activation of proinflammatory NF?B signaling. In Aim 1, I will assess the role of the Trib1:MALT1 interaction in Trib1 regulation of NF?B. I will first link Trib1 to the MALT1mediated NF?B pathway by examining the ability of Trib1 to enhance signaling in the context of MALT1 knockdown or MALT1 deficiency. Furthermore, I will assess the ability of Trib1 to potentiate MALT1induced activation of NF?B independently of stimulation. To confirm the role of the Trib1:MALT1 interaction in Trib1mediated NF?B regulation, I will generate a binding mutant of Trib1 that maintains structural integrity but abrogates the ability to bind MALT1. Both MALT1 scaffolding and protease function are regulated by ubiquitination. Given the association of Trib1 with E3 ubiquitin ligases, I hypothesize that Trib1 enhances MALT1 activity by regulating the ubiquitination status of MALT1. I will use tagged wildtype and mutant ubiquitin constructs to identify the effect of Trib1 on MALT1 ubiquitination. In Aim 2, I will determine the role of NF?B i Trib1mediated AML. I will first assess NF?B activity in Trib1mediated mouse leukemic cells, as well as in human AML samples selected for high Trib1 expression. To determine the dependence of these leukemic cells on NF?B, I will treat cells with NF?B inhibitors and examine cell survival, selfrenewal, proliferation and apoptosis. I will further assess the effect of NF?B inhibition on both induction and maintenance of Trib1mediated AML in our mouse model. I will determine the role of the Trib1:MALT1 interaction in AML by analyzing AML induction in the context of MALT1 deficiency/inhibition or by the Trib1 binding mutant developed in Aim 1. This work will provide critical insight into the pathogenesis of AML and may identify novel targets for therapeutic intervention.

描述（由申请人提供）：通过拟议的工作，我将表征的作用Tribbles同系物1（Trib 1）在NF？B信号，并确定这一作用的影响， 急性骨髓性白血病（AML）的发病机制。我们在筛选中鉴定了MALT 1作为Trib 1相互作用蛋白，以鉴定新的Trib结合伴侣。在Jurkat T细胞系中Trib 1过表达增强NF？NF中的B活性？Bresponsive报告基因测定。鉴于MALT 1在介导NF？B信号，我假设Trib 1增强NF？通过调节MALT 1活性激活B。我的实验室发表的工作将Trib 1确定为AML中的致癌基因。骨髓祖细胞中Trib 1的逆转录病毒转导在重建小鼠中诱导AML。此外，Trib 1在多种人类AML患者亚群中高度表达。我假设Trib 1通过增强促炎性NF的激活来促进AML的发病机制？B信令。在目的1，我将评估的作用Trib 1：MALT 1的相互作用，在Trib 1的NF？B。我将首先链接Trib 1的MALT 1介导的NF？B途径的研究，通过检测Trib 1在MALT 1敲低或MALT 1缺陷的情况下增强信号传导的能力。此外，我将评估Trib 1的能力，以加强MALT 1诱导的NF？B与刺激无关。为了证实Trib 1：MALT 1相互作用在Trib 1介导的NF？B调节，我将产生一个结合突变的Trib 1，保持结构完整性，但废除的能力，结合MALT 1。MALT 1支架和蛋白酶功能都受泛素化的调节。鉴于Trib 1与E3泛素连接酶的关联，我推测Trib 1通过调节MALT 1的泛素化状态来增强MALT 1的活性。我将使用标记的野生型和突变体泛素结构，以确定Trib 1对MALT 1泛素化的影响。在目标2中，我将确定NF的作用？B i Trib 1介导的AML。首先，我们来看看NF？Trib 1介导的小鼠白血病细胞以及选择高Trib 1表达的人AML样品中的B活性。确定这些白血病细胞对NF的依赖性？B，我会用NF处理细胞？B抑制剂和检查细胞存活、自我更新、增殖和凋亡。我将进一步评估NF？在我们的小鼠模型中，B抑制Trib 1介导的AML的诱导和维持。我将通过分析MALT 1缺陷/抑制背景下的AML诱导或Aim 1中开发的Trib 1结合突变体来确定Trib 1：MALT 1相互作用在AML中的作用。这项工作将为AML的发病机制提供重要的见解，并可能为治疗干预确定新的靶点。