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: 10405848
• 负责人: YI-PING LI
• 金额: $ 30.1万
• 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/10405848
• 关键词: Address; Animal Model; Ankle; Attenuated; Autoimmune Diseases; Binding Sites; Biological; Bone Diseases; Case Study; Cell Differentiation process; Cells; ChIP-seq; Chronic; Collagen Arthritis; Data; Dendritic Cells; Disease; Endogenous Factors; Genes; Goals; ITGAX gene; Immune system; Inflammation; Inflammation Mediators; Inflammatory; Joints; Knowledge; Lesion; 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; System; TNF gene; Testing; Therapeutic; Therapeutic Effect; Tissues; Transgenic Organisms; Tuberculosis; ankle joint; base; bone; cartilage degradation; conditional knockout; disability; effective therapy; gain of function; genome-wide; insight; loss of function; macrophage; 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内源信号的基础机制 减少类风湿关节炎（RA）的慢性炎症，可以为新颖的塔皮特靶标的目标提供 与炎症和组织破坏有关的疾病。 在患者的免疫系统上，会引起恶性肿瘤，肺炎和结核病 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引起的炎症和骨骼破坏的调节剂，尤其是在巨噬细胞和树突中 RA病理状况仍然难以捉摸。 巨噬细胞和树突状细胞（DC）中分化和活性的调节因子减弱Ra-dix的调节剂 局部组织破坏。 小鼠RA模型中的炎症加重。 活性形式（GNA13CA）不仅显着减少了骨骼破坏 RA的炎症表明GNA13CA可以防止RA的鼠标炎症。 RNA测序（RNA-SEQ）分析表明，Gα13可能对巨噬细胞和DC负调节 区分，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 通过TNFα或LPS刺激，我们的初步数据表明，Gα13缺乏症 基于树突状细胞和巨噬细胞中的NF-κB信号通路 我们的初步研究，我们假设Gα13是减轻炎症的主负调节剂 通过Gα13/rhoa/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。 通过表征，Gα13在类风湿关节炎（RA）中的作用在炎症和骨骼破坏中的作用 通过功能丧失的诱因（CKO）鼠标模型的型号和病理机制 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 过表达（OE）小鼠模型通过RA中的功能方法。 Gα13通过Gα13/ RA中定期炎症和骨骼破坏的机制 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。支撑研究将为您细胞的负调节提供重要的重要性 免疫系统有效靶向RA中的炎症和骨骼破坏。 可能会带来用于治疗RA和其他炎性骨疾病的陶器上级治疗方法。