Myeloid KLF2 regulation mechanisms in rheumatoid arthritis

类风湿性关节炎的髓系 KLF2 调节机制

• 批准号: 9241957
• 负责人: HIRANMOY DAS
• 金额: $ 33.66万
• 依托单位: TEXAS TECH UNIVERSITY HEALTH SCIS CENTER
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-04-01 至 2022-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9241957
• 关键词: American; Applications Grants; Arthritis; BMP2 gene; Biology; Blood; Cells; Cellular biology; Co-Immunoprecipitations; Collagen-Induced Arthritis; Complex; Disease; EP300 gene; Embryo; Exhibits; Experimental Arthritis; Experimental Designs; Family; Fibroblasts; Future; Genetic Transcription; Goals; Hematopoietic; Histologic; Histone Deacetylase; Histone Deacetylase Inhibitor; Human; Immune; Immune system; In Vitro; Infiltration; Inflammation; Inflammatory; Joints; K/BxN model; Knockout Mice; Knowledge; Kruppel-like transcription factors; MMP9 gene; Mediating; Molecular; Mus; Myelogenous; Myeloid Cell Activation; Myeloid Cells; NF-kappa B; Osteoclasts; PCAF gene; Pathogenesis; Pathway interactions; Patients; Pharmacology; Play; Process; Radiology Specialty; Recruitment Activity; Regulation; Reporting; Rheumatoid Arthritis; Role; Serum; Structure; TNFSF11 gene; Therapeutic; Tissues; Transcription Factor AP-1; Zinc Fingers; base; cathepsin K; chromatin immunoprecipitation; disorder control; gain of function; gene function; gene repression; in vivo; interest; loss of function; macrophage; member; microCT; monocyte; novel; osteoclastogenesis; peripheral blood; public health relevance; response; soft tissue; transcription factor

 DESCRIPTION (provided by applicant): The long-term objective of this proposal is to investigate the role of zinc finger transcriptional factor, Kruppel like factor-2 (KLF2) in the regulation of osteoclastogenesis and rheumatoid arthritis (RA)-related pathogenesis. Among the cellular components monocytes play major roles in mediating progression of RA. The monocytes migrate from the peripheral blood to the arthritic joint tissues and secrete pro-inflammatory factors, and these factors in turn mediate inflammation and recruit other immune cells, which mediate differentiation of monocytes towards osteoclasts. Since it has been reported that KLF2 can induce quiescence to the immune cells and also inactivate monocytes in response to stimulation (active monocytes are required to promote osteoclastogenesis) and because our preliminary results indicate severe arthritic joint changes in KLF2 hemizygous mice (KLF2 homozygous knock out mice are embryonically lethal), we therefore hypothesize that this transcription factor may inhibit osteoclastogenesis and thereby, pathogenesis of RA. Aim1 will determine the importance of myeloid KLF2 in regulating the pathogenesis in RA. Experimental Design: Conditional gain and loss of gene function approach will be undertaken to determine the role of KLF2 in the regulation of pathogenesis in vivo. Complementary histologic, histomorphometric, immunohistochemical, and radiologic (micro CT) studies will be used to evaluate extent of the disease. Human monocytes and joint tissues from RA and healthy controls will be studied to confirm findings. Aim 2 will examine the bases for KLF2's ability to regulate osteoclastogenesis. Experimental Design: Gain- and loss-of-function approaches will be used to determine the role of KLF2 in osteoclastic differentiation. Additionally, the cross talk between KLF2 and osteoclastogenesis markers (NFATc1, NFκB, Cathepsin K and MMP9) and osteoblastogenesis markers (RANK/RANKL, OPG, BMP2/4, and Runx2) will be determined. Human monocytes from RA and healthy controls will also be studied to confirm findings. Aim 3 will investigate whether the effects of pharmacological compound on arthritis and osteoclastogenesis are KLF2 dependent. We found that a group of pharmacological compound, such as HDAC inhibitor (HDACi) induces KLF2 expression in myeloid cells. Experimental Design: We will verify if HDACi induce KLF2 in myeloid cells in mice. Next, we will determine if the HDACi-mediated regulation of inflammation and osteoclastogenesis is KLF2- dependent using various molecular approaches (structure-functions studies, co-immunoprecipitation, and ChIP assays). The knowledge generated from this study will not only identify a novel endogenous regulator of osteoclastogenesis in RA, but also indicate newer and effective strategies to control this disease.

 描述（由适用提供）：该提案的长期目标是研究锌指转录因子，Kruppel（例如因子-2（KLF2））在破骨细胞生成和类风湿关节炎（RA）相关发病机制中的作用。在细胞成分中，单核细胞在介导RA的进展中起着重要作用。单核细胞从外周血迁移到关节炎关节组织和秘密促炎性因素，这些因素又又介绍了培养基的炎症并募集了其他免疫细胞，从而培养了单核细胞向破骨细胞的分化。 Since it has been reported that KLF2 can induce quiescence to the immunocytes and also inactivate monocytes in response to stimulation (active monocytes are required to promote osteoclastogenesis) and because our preliminary results indicate severe arthritik joint changes in KLF2 hemizygous mice (KLF2) homozygous knock out mice are embryonically lethal), we therefore假设该转录因子可能抑制破骨细胞生成，从而抑制RA的发病机理。 AIM1将确定髓样KLF2在确定RA的发病机理中的重要性。实验设计：将进行条件增益和基因功能方法的丧失，以确定KLF2在体内发病机理调节中的作用。互补的组织学，组织形态计量学，免疫组织化学和放射学（微CT）研究将用于评估疾病的程度。将研究RA和健康对照的人类单核细胞和关节时间，以确认发现。 AIM 2将检查KLF2调节破骨细胞生成的能力的碱基。实验设计：将使用功能丧失方法来确定KLF2在整界碎屑分化中的作用。此外，串口 在KLF2和破骨细胞生成标志物（NFATC1，NFκB，组织蛋白酶K和MMP9）和骨碎片发生标记（RANK/RANKL，OPG，BMP2/4和RUNX2）之间。 RA和健康对照组的人类单核细胞也将进行研究以确认发现。 AIM 3将研究药物化合物对关节炎和破骨构成的作用是否取决于KLF2。我们发现一组药物化合物，例如HDAC抑制剂（HDACI）会诱导髓样细胞中的KLF2表达。实验设计：我们将使用各种分子方法（结构功能研究，共免疫沉淀和芯片分析）验证HDACI介导的感染和破骨细胞生成的调节是KLF2-依赖性的。这项研究产生的知识不仅将确定RA中破osteogogenesogeny的新型内源性调节剂，而且还表明了控制这种疾病的新策略。