Pim1 kinase coordinates PPAR gamma pathway and mitochondrial function to mediate pro-atherogenic responses in macrophages

Pim1 激酶协调 PPAR gamma 通路和线粒体功能，介导巨噬细胞中的促动脉粥样硬化反应

• 批准号: 10591599
• 负责人: WEIGUO CUI
• 金额: $ 41.69万
• 依托单位: MEDICAL COLLEGE OF WISCONSIN
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-01 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10591599
• 关键词: Ablation; Adopted; Atherosclerosis; Attenuated; Binding; Biochemical; Bone Marrow; CD36 gene; Cause of Death; Cells; Collaborations; Coupled; Data; Development; Diet; Disease; Dynamin; Dyslipidemias; Fatty Acids; Feedback; Foam Cells; Genetic; Genetic Transcription; Goals; Immune; Immunologics; Infusion procedures; Interleukin-6; Intracellular Accumulation of Lipids; Knowledge; Ligands; Link; Lipids; Macrophage; Macrophage Activation; Mediating; Metabolic Pathway; Mitochondria; Modeling; Molecular; Morphology; Mus; Nuclear Hormone Receptors; Oxidative Phosphorylation; Oxidative Stress; PIM1 gene; PPAR gamma; Pathway interactions; Phenotype; Phosphotransferases; Play; Process; Production; Protein-Serine-Threonine Kinases; Proteins; Publishing; Resistance; Role; Serine; Signal Transduction; Site; Stat3 protein; Surface; Testing; Therapeutic Intervention; Up-Regulation; fatty acid metabolism; fatty acid oxidation; immune activation; in vivo; inhibitor; lipid metabolism; metabolic abnormality assessment; metabolic profile; mitochondrial dysfunction; monocyte; new therapeutic target; novel; novel therapeutic intervention; oxidized low density lipoprotein; pharmacologic; programs; response; scavenger receptor; transcription factor; uptake

Atherosclerosis (AS) remains the leading cause of death world-wide and it is often associated with dyslipidemia, oxidative stress and mitochondrial dysfunction. Macrophage is a type of innate immune cell that plays a critical role in the development of AS. Unrestricted uptake of oxidized LDL (oxLDL) by macrophages leads to accumulation of lipid intracellularly and foam cell formation, which is a hallmark of early stages of AS. OxLDL uptake is mainly mediated by CD36, a scavenger receptor highly expressed on the macrophage surface. One major problem is that oxLDL leads to up-regulation of CD36 expression through a transcription factor PPARγ, resulting in a positive feedback mechanism to further enhance CD36-mediated oxLDL uptake. Defining the novel regulator of this process is the central goal of this multi-PI proposal. Recent published and preliminary studies showed that Pim1, a conserved serine/threonine kinase regulates CD36 transcription. In addition, genetic ablation of pim1 gene in macrophages resulted in a reduction in PPARγ pathway as well as the downstream targets involved in fatty acid metabolism and mitochondrial oxidative phosphorylation. We thus hypothesized that Pim1 kinase coordinates PPARγ activation/CD36 expression and mitochondrial functions to regulate fatty acid metabolism and immune activation in macrophages. Continuous stimulation of Pim1 kinase in macrophages contributes to pro-atherogenic phenotypes and AS. Specific aim 1 will determine the molecular mechanism by which Pim1 kinase in macrophages coordinates PPARγ activation/CD36 expression and mitochondrial function to modulate fatty acid metabolism. We will use a combination of genetically modified macrophages, biochemical, immunological and ex vivo cell metabolic studies to determine the mechanisms by which Pim1 kinase regulates fatty acid metabolism through the PPARγ/CD36 pathway; and to determine the mechanisms by which Pim1 kinase regulates mitochondria morphology through Drp-1 and define the impact on mitochondria fatty acid oxidation, oxidative phosphorylation and ROS production. Aim 2 will test the hypothesis that inactivating Pim1 kinase in vivo suppresses the development of AS. Using the genetic pim1 ablation model and minipump infusion of the Pim inhibitor AZD1208, we aim to test the hypothesis that Pim1 activity is indispensible for diet-induced AS in mice; and to test the hypothesis that pharmacologic inhibition of Pim1 suppresses PPARγ/CD36 pathway and reprograms macrophage mitochondria toward ROS production under atherogenic conditions; and test the hypothesis that Pim1 regulates bone marrow-monocyte-macrophage differentiation lineage under atherogenic conditions. By elucidating the molecular mechanisms by which Pim1 kinase coordinates fatty acid metabolism and mitochondrial functions to control macrophage activation under atherogenic conditions, we expect to gain crucial knowledge on novel lipid metabolism regulators and provide new treatment strategies against AS.

动脉粥样硬化（AS）仍然是世界范围内死亡的主要原因，它往往与 血脂异常、氧化应激和线粒体功能障碍。巨噬细胞是一种先天免疫细胞， 在AS的发展中起着关键作用。巨噬细胞对氧化型低密度脂蛋白（oxLDL）的无限制摄取 导致细胞内脂质积聚和泡沫细胞形成，这是AS早期阶段的标志。 氧化低密度脂蛋白的摄取主要由巨噬细胞上高表达的清道夫受体CD 36介导 面一个主要的问题是oxLDL通过转录上调CD 36的表达， 因子PPARγ，导致正反馈机制进一步增强CD 36介导的oxLDL摄取。 定义这个过程的新调节器是这个多PI提案的中心目标。最近出版的和 初步研究表明，Pim 1，一种保守的丝氨酸/苏氨酸激酶，调节CD 36的转录。在 此外，巨噬细胞中pim 1基因的基因切除导致了PPARγ途径的减少， 下游靶点涉及脂肪酸代谢和线粒体氧化磷酸化。我们因此 假设Pim 1激酶协调PPARγ活化/CD 36表达和线粒体功能， 调节巨噬细胞的脂肪酸代谢和免疫激活。Pim 1激酶的连续刺激 在巨噬细胞中的表达有助于促动脉粥样硬化表型和AS。具体目标1将决定 巨噬细胞中Pim 1激酶协调PPARγ活化/CD 36表达的分子机制 以及线粒体调节脂肪酸代谢的功能。我们将结合基因 修饰的巨噬细胞，生物化学，免疫学和离体细胞代谢研究，以确定 Pim 1激酶通过PPARγ/CD 36途径调节脂肪酸代谢的机制;以及 确定Pim 1激酶通过Drp-1调节线粒体形态的机制， 定义对线粒体脂肪酸氧化、氧化磷酸化和ROS产生的影响。目的2 将检验体内Pim 1激酶失活抑制AS发展的假设。使用 我们的目标是通过基因pim 1消融模型和微型泵输注Pim抑制剂AZD 1208来测试 假设Pim 1活性对于小鼠中饮食诱导的AS是必不可少的;并检验假设， Pim 1的药理学抑制抑制了PPARγ/CD 36通路并重新编程巨噬细胞 在致动脉粥样硬化条件下，线粒体对ROS的产生;并检验Pim 1 在动脉粥样硬化条件下调节骨髓-单核细胞-巨噬细胞分化谱系。通过 阐明Pim 1激酶协调脂肪酸代谢的分子机制， 线粒体的功能，以控制巨噬细胞活化动脉粥样硬化条件下，我们希望获得 新的脂质代谢调节剂的关键知识，并提供新的治疗策略，对AS。