Suppressing Inflammation by Blocking IKK Oligomer

通过阻断 IKK 寡聚物抑制炎症

基本信息

批准号：
10573218
负责人：
GOURISANKAR GHOSH
金额：
$ 52.93万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN DIEGO
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-02-15 至 2027-01-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10573218
关键词：
Affect Architecture Arthritis Autoimmune Autoimmunity Binding Biophysics Black race Cartoons Catalytic Domain Cells Chemicals Collagen Arthritis Complex Coupled Defect Disease Event Failure Goals Heart IkappaB kinase In Vitro Industrialization Infection Inflammation Inflammatory Innate Immune Response Light Link Malignant Neoplasms Modeling Molecular Conformation Molecular Weight Mutation NF-kappa B Pathogenicity Peptides Pharmaceutical Preparations Phosphorylation Phosphotransferases Play Point Mutation Predisposition Process Protein Kinase Regulation Research Personnel Role Scaffolding Protein Serine Signal Pathway Signal Transduction Site Specificity Stimulus Syndrome Testing Therapeutic Transcriptional Activation Ubiquitin Zinc Fingers adaptive immune response dimer drug development efficacy testing experimental study extracellular fighting in vivo inhibitor intermolecular interaction monomer mouse model mutant novel strategies potency testing protein complex protein protein interaction receptor response scaffold systemic inflammatory response transcription factor

项目摘要

Uncontrolled activation of the IkappaB kinase (IKK), the key cellular regulator of NF-κB, causes a variety of disorders, including susceptibility to pathogenic infection, autoimmunity, inflammation-induced malignancies, and inflammatory syndromes. However, targeting the IKK-NF-kB signaling pathway has not yielded any new strategies for fighting inflammatory illnesses. The lack of understanding of how IKK becomes activated in response to various stimuli is the fundamental reason for our failure to exploit this unambiguous target. The IKK complex is made up of three subunits, catalytic IKK1 (also known as IKKa) and IKK2/b kinases, which form a heterodimer, and the dimeric scaffolding protein NEMO (NF-κB Essential Modulator), which is stably bound to the heterodimer. In vitro and in cells, this fundamental tetrameric unit of the IKK complex (IKK1:IKK2:NEMO2), emerges as significantly higher molecular weight multimers. The catalytic activation of IKK2, referred to as canonical signaling, requires linear (M1-linked), K63-linked, or mixed poly-ubiquitin chains (Ub-chain) that engage noncovalently with the NEMO subunits. The mechanism by which this binding information is transferred from NEMO:Ub-chain interaction to yield IKK2 subunit phosphorylation and subsequent activation of IKK is unknown. We hypothesize that multimerization via dimer-dimer interaction is required for IKK2 activation and that the tetramer interface stabilizes the native IKK complex, allowing NEMO to undergo conformational changes upon binding to the Ub-chain. The kinase domain of the IKK2 subunit is activated as a result of structural alterations in NEMO. Disease-causing NEMO mutations or mutations at the tetramer interface do not support the assembly and Ub-chain-dependent NEMO conformational changes required for IKK activation. In support of our hypothesis, we have already shown that a short peptide segment derived from NEMO interacts with IKK2 in a signal-dependent manner and that IKK multimerization requires short homologous peptide segments within IKK1 and IKK2. Under this proposal, we will achieve the following specific aims: AIM 1. We will characterize the dimer-dimer interface which is required for IKK multimerization. We will use disease-causing NEMO mutants and IKK multimerization-defective mutants to test how the structural plasticity of NEMO is linked to IKK2 activation. AIM 2. We will determine if the IKK1- and IKK2-derived peptides disrupt dimer-dimer interaction and IKK2 activation in cells and in vivo. We will test if the IKK-derived peptides independently and in combination with the NEMO-derived peptide can ameliorate systemic inflammation and collagen-induced arthritis in mouse models.

NF-κB的关键细胞调节剂IKAPPAB激酶（IKK）的不受控制激活会引起多种疾病，包括对病原感染，自身免疫性，炎症引起的恶性肿瘤和炎症综合症的易感性。但是，针对IKK-NF-KB信号通路途径并未产生与炎症性疾病作斗争的任何新策略。缺乏对IKK如何因各种刺激而被激活的理解是我们未能利用这一明确目标的基本原因。 IKK 复合物由三个亚基IKK1（也称为IKKA）和IKK2/B激酶组成，它们形成异二聚体，以及二聚体脚手架蛋白Nemo（NF-κB必需调节剂），它们稳定地结合了与杂二聚体。在体外和细胞中，这种IKK复合物的基本四聚体单位（IKK1：IKK2：NEMO2）出现的分子量多聚体明显更高。 IKK2的催化激活，称为规范信号，需要线性（M1连接），K63连接或混合多泛素链（UB链）与Nemo亚基无比互动。这些结合信息从NEMO：UB链相互作用传递以产生IKK2亚基磷酸化和随后的IKK激活的机制尚不清楚。我们假设IKK2激活和四聚体界面稳定了天然IKK复合物，从而使Nemo在与UB链结合时会发生构象变化。 IKK2亚基的激酶结构域由于NEMO的结构改变而被激活。在四聚体界面处引起疾病的NEMO突变或突变不支持IKK激活所需的组装和UB链依赖性的NEMO构象变化。为了支持我们的假设，我们已经表明，源自Nemo的短肽段与IKK2相互作用信号依赖性的方式和IKK多层化需要在IKK1和IKK2中短的同源肽段。在此提案下，我们将实现以下特定目标：AIM 1。我们将表征IKK多层化所需的二聚体二聚体界面。我们将使用引起疾病的NEMO突变体和IKK多层化缺陷突变体来测试NeMo的结构可塑性如何与IKK2激活有关。 AIM 2。我们将确定IKK1-和IKK2衍生的Petides是否破坏了细胞和体内IKK2的二聚体二聚体相互作用和IKK2激活。我们将测试IKK衍生的Petides是否独立并与Nemo衍生的Petides结合使用可以改善小鼠模型中的全身感染和胶原蛋白诱导的关节炎。