Characterizing the negative signaling in dendritic cells and macrophages to attenuate inflammation and bone destruction in rheumatoid arthritis

表征树突状细胞和巨噬细胞中的负信号传导以减轻类风湿性关节炎的炎症和骨质破坏

• 批准号: 10616608
• 负责人: YI-PING LI
• 金额: $ 33.44万
• 依托单位: TULANE UNIVERSITY OF LOUISIANA
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-02-15 至 2024-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10616608
• 关键词: Animal Model; Ankle; Arthritis; Attenuated; Autoimmune Diseases; Binding Sites; Biological; Bone Diseases; Case Study; Cell Differentiation process; Cells; ChIP-seq; Chronic; Collagen Arthritis; Cre lox recombination system; Data; Dendritic Cells; Disease; Endogenous Factors; Genes; Goals; ITGAX gene; Immune system; Inflammation; Inflammation Mediators; Inflammatory; Joints; Knockout Mice; Knowledge; Lesion; Macrophage; Malignant Neoplasms; Mediating; Modeling; Molecular; Mus; NF-kappa B; Pathologic; Pathway interactions; Patients; Peripheral; Phenotype; Phosphorylation; Pneumonia; Proto-Oncogene Proteins c-akt; Regulation; Research; Rheumatoid Arthritis; Role; STAT3 gene; Signal Pathway; Signal Transduction; Synovitis; TNF gene; Testing; Therapeutic; Therapeutic Effect; Tissues; Transgenic Organisms; Tuberculosis; ankle joint; bone; cartilage degradation; conditional knockout; disability; effective therapy; gain of function; genome-wide; insight; loss of function; mouse model; new therapeutic target; overexpression; promoter; transcriptome sequencing

The goal of this proposal is to understand the mechanisms underlying how negative endogenous signaling reduces chronic inflammation in rheumatoid arthritis (RA), which can provide novel therapeutic targets for diseases related to inflammation and tissue destruction. Current therapies for RA have serious negative effects on the patients’ immune system, and can cause malignancies, pneumonia, and tuberculosis. Positive regulators of inflammation in RA (such as NF-κB) have been well characterized, however, the negative endogenous regulators of RA-induced inflammation and bone destruction, especially in macrophages and dendritic cells in RA pathological condition remain elusive. The proposed study will characterize the negative endogenous regulators of differentiation and activity in macrophages and dendritic cells (DCs) that attenuate RA-induced inflammation and tissue destruction. We noted that Gα13 deficiency in macrophages or dendritic cells aggravated inflammation in mouse RA model. AAV mediated overexpression of local Gna13 constitutively active form (Gna13CA) not only significantly reduced bone destruction, but also dramatically inhibited inflammation in RA, indicating that Gna13CA could protect against inflammation in a mouse model of RA. Our RNA sequencing (RNA-seq) analysis showed that Gα13 may negatively regulate macrophage and DC differentiation and activation and ChIP sequencing (ChIP-seq) analysis showed the enrichment of NF-κB motif site binding on the genome-wide promoters of the genes related to inflammation in Gα13 deficient macrophages with TNFα or LPS stimulations. Our preliminary data showed that Gα13 deficiency promotes Akt activity, STAT3 phosphorylation and NF-κB signaling pathway activation in dendritic cells and macrophages. Based on our preliminary studies, we hypothesize that Gα13 is a master negative regulator that attenuates inflammation and bone destruction associated with rheumatoid arthritis through the Gα13/RhoA/AKT/IKK/NF-κB pathway in macrophages and dendritic cells. Three specific aims are proposed to test our hypothesis. In Aim 1, we will determine the role of Gα13 in inflammation and bone destruction in rheumatoid arthritis (RA) by characterizing the phenotypes and pathomechanism of conditional knockout (CKO) mouse models via loss-of-function approach in RA. In Aim 2, we will define the roles and therapeutic effects of Gα13CA in inflammation and bone destruction in RA by characterizing the phenotypes and pathomechanism of conditional transgenic overexpression (OE) mouse models via gain-of-function approach in RA. We will characterize the molecular mechanism by which Gα13 regulates inflammation and bone destruction in RA through Gα13/ RhoA/AKT/IKK/NF-κB signaling, STAT3 signaling, and TLRs signaling pathways in macrophages and dendritic cells in Aim 3. The proposed study will provide important insights into the negative regulation of the cells of the immune system to effectively target inflammation and bone destruction in RA. Knowledge gained from this study may bring forth potential superior therapeutic means for treating RA and other inflammatory bone diseases.

该提案的目的是了解负面内源信号的基础机制 减少类风湿关节炎（RA）中的慢性炎症，可以为您提供新的治疗靶点 与感染和组织破坏有关的疾病。当前的RA疗法具有严重的负面影响 在患者的免疫系统上，可能导致恶性肿瘤，肺炎和结核病。积极的监管机构 但是，RA中的炎症（例如NF-κB）已得到很好的表征，但是内源性为负 RA诱导的注射和骨骼破坏的调节剂，尤其是在巨噬细胞和树突状细胞中 RA病理状况仍然难以捉摸。拟议的研究将表征负内源性 巨噬细胞和树突状细胞（DC）分化和活性的调节剂，可减弱RA诱导的 炎症和组织破坏。我们注意到巨噬细胞或树突状细胞中的Gα13缺乏 小鼠RA模型中的聚集炎症。 AAV介导的局部GNA13的过表达 活动形式（GNA13CA）不仅显着减少了骨骼破坏，而且还大大抑制了 RA中的炎症，表明GNA13CA可以预防RA小鼠模型中的炎症。我们的 RNA测序（RNA-SEQ）分析表明，Gα13可能对巨噬细胞和DC负调节 分化，激活和芯片测序（CHIP-SEQ）分析显示了NF-κB基序的富集 位点结合在与Gα13缺陷巨噬细胞中与炎症有关的基因的全基因组启动子上结合 带有TNFα或LPS刺激。我们的初步数据表明，Gα13缺乏促进了Akt活性，STAT3 树突状细胞和巨噬细胞中的磷酸化和NF-κB信号通路激活。基于 我们的初步研究，我们假设Gα13是一种减弱注射的主负调节剂 通过Gα13/Rhoa/akt/ikk/ikk/nf-κB途径与类风湿关节炎相关的骨骼破坏 巨噬细胞和树突状细胞。提出了三个特定目标来检验我们的假设。在AIM 1中，我们将 通过表征，Gα13在类风湿关节炎（RA）中的作用在炎症和骨破坏中的作用 有条件敲除（CKO）小鼠模型的表型和病理机制通过功能丧失 RA的方法。在AIM 2中，我们将定义Gα13CA在炎症和骨骼中的作用和治疗作用 通过表征有条件转基因的表型和病理机制，通过表征RA的破坏 通过RA中的功能获取方法过表达（OE）模型。我们将表征分子 Gα13通过Gα13/ RA中调节注射和骨骼破坏的机制 rhoa/akt/ikk/ikk/nf-κB信号传导，STAT3信号传导和TLRS信号传导途径和树突状 AIM 3中的细胞。拟议的研究将为细胞的负调节提供重要的见解。 免疫系统有效靶向RA中的注射和骨骼破坏。从这项研究中获得的知识 可能会带来潜在的上级治疗手段，以治疗RA和其他炎症性骨骼疾病。