The role of smooth muscle PPAR gamma in neointima formation

平滑肌 PPAR γ 在新内膜形成中的作用

• 批准号: 8458140
• 负责人: Seth Bernat Furgeson
• 金额: $ 13.04万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-04-18 至 2016-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8458140
• 关键词: 2,4-thiazolidinedione; Affect; Agonist; Alleles; Angioplasty; Animal Model; Anti-Inflammatory Agents; Anti-inflammatory; Antineoplastic Agents; Arterial Injury; Atherosclerosis; Bexarotene; Blood Vessels; Bone Marrow; Bone Marrow Transplantation; CCL2 gene; Cell Proliferation; Cells; Cellular biology; Complication; Contractile Proteins; Data; Drug usage; Family; Galactosidase; Gene Expression; Gene Targeting; Genetic Transcription; Goals; Human; In Vitro; Inflammatory; Injury; Interleukin-6; Knock-out; Knockout Mice; Label; Ligands; Measures; Mediating; Mediator of activation protein; MicroRNAs; Modeling; Morbidity - disease rate; Mus; Nuclear Receptors; PPAR gamma; Peroxisome Proliferator-Activated Receptors; Phenotype; Pioglitazone; Process; Production; Proliferating; Publishing; RXR; Relative (related person); Reporter; Response Elements; Role; Signal Pathway; Site; Smooth Muscle; Smooth Muscle Myocytes; Stromal Cell-Derived Factor 1; Testing; Thiazolidinediones; Time; Tissues; Toxic effect; Vascular Diseases; activating transcription factor; base; cell motility; cell type; chemokine; cytokine; design; diabetic; effective therapy; femoral artery; in vivo; macrophage; member; migration; neointima formation; public health relevance; response; restenosis; rosiglitazone

DESCRIPTION (provided by applicant): PROJECT SUMMARY The goal of this project is to define the role of peroxisome proliferator-activated receptor (PPAR)3 activation in neointima formation. Neointima formation occurs frequently after angioplasty and causes significant morbidity; vascular smooth muscle cells (SMCs) are key cells during neointima formation. We will study the effects of two clinically available agents on SMC biology and neointima formation. PPAR3 is a ligand-activated nuclear receptor that has been shown to have beneficial effects on vascular disorders. We will compare the effects of two agents: pioglitazone (activates PPAR3 only) and bexarotene (an RXR agonist which activates PPAR3, PPAR1, PPAR4, LXR, and FXR). Our hypothesis is that PPAR3 activation specifically in smooth muscle cells (SMC) will reduce neointima formation by decreasing resident SMC migration and proliferation as well as SMC-derived chemokine production and subsequent recruitment of bone marrow-derived cells. We believe both pioglitazone and bexarotene will be effective but bexarotene may be more effective due to activation of other nuclear receptors. In Aim One, we will compare the effects of pioglitazone to bexarotene on SMCs. We will measure changes in proliferation, cytokine production, and smooth muscle gene expression. In Aim Two, we will determine if the agents affect levels of microRNAs crucial to maintaining SMC phenotype, such as miR- 143, miR-145, and miR-221. In Aim Three, we will examine the effects of the agents in vivo during femoral artery wire injury. To track recruitment of bone-marrow derived cells to the site of arterial injury, all mice will receive bone marrow transplants from a GFP positive donor. After wire injury, mice will be analyzed at multiple time points. Along with neointima size, we will measure production of chemokines (IL-6, MCP-1, SDF-11, and KC), recruitment of bone marrow-derived cells and macrophages, and cellular proliferation. We also plan to study the role of PPAR3 activation specifically in smooth muscle cells during neointima formation. Using an inducible tissue-specific knockout model, we will deplete PPAR3 in smooth muscle cells after mice have received bone marrow transplants from GFP positive donors. Inducible PPAR3 knockout mice and control mice will receive therapy with either pioglitazone, bexarotene or control and be subjected to femoral artery wire injury. At multiple time points, neointima size will again be measured. All mice used in Aim 3B will have the R26R reporter allele in which smooth muscle cells are labeled with 2-galactosidase. Since we can measure both bone marrow derived and resident SMCs, we will determine the relative contribution each cell type makes to the neointima. We will also be able to determine if PPAR3 specifically in smooth muscle cells mediates the effects of bexarotene or pioglitazone.

描述（由申请人提供）：项目摘要该项目的目的是定义过氧化物酶体增殖物激活受体（PPAR）3在新内膜形成中的激活。新内膜形成在血管成形术后经常发生，并引起明显的发病率。血管平滑肌细胞（SMC）是新内膜形成期间的关键细胞。我们将研究两种临床上可用药物对SMC生物学和新内膜形成的影响。 PPAR3是一种配体激活的核受体，已证明对血管疾病具有有益的作用。我们将比较两种药物的效果：吡格列酮（仅激活PPAR3）和贝克索烯（激活PPAR3，PPAR1，PPAR4，LXR和FXR）的RXR激动剂。我们的假设是，PPAR3在平滑肌细胞（SMC）中的特异性激活将通过减少居民SMC迁移和增殖以及SMC衍生的趋化趋化因子产生以及随后募集骨髓衍生的细胞来减少新内膜形成。我们认为，吡格列酮和贝克索烯都会有效，但是由于其他核受体的激活，贝沙罗汀可能更有效。在AIM中，我们将比较吡格列酮对贝克索烯对SMC的影响。我们将测量增殖，细胞因子产生和平滑肌基因表达的变化。在目标两个方面，我们将确定药物是否影响microRNAS的水平，对维持SMC表型至关重要，例如miR-143，miR-145和miR-221。在AIM三中，我们将检查股动脉线损伤期间体内药物的作用。为了跟踪骨髓衍生细胞到动脉损伤部位的募集，所有小鼠将从GFP阳性供体中接收骨髓移植。电线损伤后，将在多个时间点分析小鼠。随着新内膜大小，我们将测量趋化因子（IL-6，MCP-1，SDF-11和KC）的产生，骨髓来源的细胞和巨噬细胞的募集以及细胞增殖。我们还计划在新内膜形成过程中专门研究PPAR3激活在平滑肌细胞中的作用。使用诱导组织特异性敲除模型，在小鼠接受GFP阳性供体的骨髓移植后，我们将在平滑肌细胞中耗尽PPAR3。可诱导的PPAR3敲除小鼠和对照小鼠将接受吡格列酮，贝克索烯或对照的治疗，并遭受股动脉线损伤。在多个时间点，将再次测量新内膜大小。 AIM 3B中使用的所有小鼠均具有R26R报告基因等位基因，其中平滑肌细胞用2-半乳糖苷酶标记。由于我们可以测量衍生的骨髓和常驻SMC，因此我们将确定每种细胞类型对新内膜的相对贡献。我们还将能够确定平滑肌细胞中的PPAR3是否介导了北卡罗烯或吡格列酮的作用。