INTERACTION OF MESENCHYMAL AND MYELOID FIBROBLASTS IN INFLAMMATORY-BASED FIBROSIS IN THE AGING HEART

间充质和骨髓成纤维细胞在衰老心脏炎症性纤维化中的相互作用

• 批准号: 9790895
• 负责人: KATARZYNA A. CIESLIK
• 金额: $ 48.36万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-30 至 2022-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9790895
• 关键词: Activation Analysis; Acute; Adoptive Transfer; Age; Aging; Binding; Biology; CCL2 gene; CD209 gene; Cardiac; Cardiac Function Study; Cells; Collagen; Cytometry; Data; Defect; Dendritic Cells; Deposition; Development; Disease; Elderly; Electron Microscopy; Epidemic; Event; Fibroblasts; Fibronectins; Fibrosis; Functional disorder; Generations; Goals; Heart; Heart Injuries; Human; Impairment; In Vitro; Individual; Infiltration; Inflammation; Inflammatory; Inflammatory Infiltrate; Injury; Knock-out; Laboratories; Leukocytes; Ligands; Link; Lymphocyte; Lymphocyte antigen CD50; Magnetic Resonance Imaging; Mass Spectrum Analysis; Measurement; Mechanics; Mesenchymal; Mesenchymal Stem Cells; Mission; Modeling; Molecular; Mononuclear Leukocytes; Mus; Myelogenous; Myeloid Cells; Nature; Pathologic; Pathway interactions; Phenotype; Phosphorylation; Population; Process; Protein Array; Protein Biosynthesis; Public Health; Research; Role; Signal Pathway; Signal Transduction; T-Lymphocyte; Tissues; Transforming Growth Factor beta; Transforming Growth Factor beta Receptors; Work; age related; aged; aging population; base; cell type; coronary fibrosis; cytokine; effective therapy; heart function; human tissue; improved; in vivo; inhibitor/antagonist; innovation; interstitial; macrophage; monocyte; mouse model; non-invasive monitor; receptor; response; stemness

In 2030, elderly people will represent 20% of the US population. The focus of our work is on the mechanism(s) causing fibrosis and diastolic dysfunction, which has become epidemic in the aging population. Our overall objective is to examine cellular and molecular interactions between fibroblasts of two developmental origins (mesenchymal and myeloid) and potential rescue approaches to reduce diastolic dysfunction and cardiac fibrosis in the aging heart. The central hypothesis arose from our data in which we identified the signaling abnormalities in mesenchymal fibroblasts that directly contribute to their aberrant activation resulting in fibrosis and stimulation of monocyte influx into the heart. Infiltrating monocytes polarize into myeloid fibroblasts and further contribute to mesenchymal fibroblast pathological activation. To examine the contribution of each fibroblast type and their interaction, we will use specific inhibitors such as 1) AICAR, an AMPK activator that inhibits the pathologically upregulated Erk pathway in mesenchymal fibroblasts and thereby reduces collagen and fibronectin levels in the aging heart and 2) DCSL-1, which specifically binds to DC-SIGN, a receptor found exclusively in leukocytes. We have found that in vivo DCSL-1 treatment improves cardiac function and decreases the number of M1 proinflammatory macrophages and myeloid fibroblasts in the heart as well as reducing collagen deposition. In SA1, we will determine the mechanism by which defective mesenchymal fibroblasts contribute to interstitial fibrosis in the aging heart and examine the role of an AMPK activator in reversing these defects. We will examine the molecular mechanism by which pathological matrix deposition occurs in aged mouse and human tissues using electron microscopy, mass spectrometry and signaling pathway activation analyses. We will also determine the impact of in vivo and in vitro AICAR on these processes. Finally, the changes in cardiac fibrosis and function after in vivo AICAR treatment in mice will be monitored by noninvasive MRI, Echo and Doppler measurements. In SA2, we will investigate the role of the inflammatory infiltrate in fibrosis and in the altered mesenchymal fibroblast phenotype by use of a myeloid- specific inhibitor, DCSL1. We will compare aged mice treated with DCSL-1 with age-matching controls and study infiltrating leukocytes via noninvasive cell tracking using MRI, analyze infiltrating monocytes and lymphocytes via mass cytometry and identify cytokines secreted by them via protein array. Furthermore, the role of DCSL-1 on T lymphocyte populations will be investigated using adoptive transfer. For in vitro studies the effect of secreted cytokines on mesenchymal fibroblasts of mouse and human origin will be analyzed as well. Finally, matrix analysis and cardiac function study to compare DCSL-1 and control treated aged mice will be done as in SA1. This approach is innovative and significant because it will allow us to link changes in cardiac function to molecular and cellular responses. Furthermore, our Preliminary Data shows a correlation of the mouse model with the human aging heart, where we observed the same defects.

到2030年，老年人将占美国人口的20%。我们工作的重点是机制 引起纤维化和舒张功能障碍，这在老龄人口中已成为流行病。我们的整体 目的是研究两种发育来源的成纤维细胞之间的细胞和分子相互作用 （间质和骨髓）和潜在的补救方法，以减少舒张功能障碍和心脏 老化心脏的纤维化核心假设来自我们的数据，我们在这些数据中发现了 间充质成纤维细胞的异常，直接导致其异常活化，导致纤维化 和刺激单核细胞流入心脏。单核细胞浸润骨髓成纤维细胞， 进一步促进间充质成纤维细胞病理活化。为了检查每个人的贡献， 成纤维细胞类型和它们的相互作用，我们将使用特异性抑制剂，如1）AICAR，一种AMPK激活剂， 抑制间充质成纤维细胞中病理性上调的Erk途径，从而减少胶原蛋白 和纤维连接蛋白水平，以及2）DCSL-1，它特异性结合DC-SIGN，一种发现的受体 仅限于白细胞。我们已经发现体内DCSL-1治疗改善了心脏功能， 减少心脏中M1促炎性巨噬细胞和骨髓成纤维细胞的数量， 减少胶原沉积。在SA 1中，我们将确定缺陷间充质细胞 成纤维细胞在衰老心脏中促进间质纤维化，并研究AMPK激活剂在 弥补这些缺陷。我们将研究病理性基质沉积 发生在老年小鼠和人体组织使用电子显微镜，质谱和信号 途径活化分析。我们还将确定体内和体外AICAR对这些细胞的影响。 流程.最后，将在小鼠体内AICAR治疗后观察心脏纤维化和功能的变化。 通过无创MRI、回波和多普勒测量进行监测。在SA 2中，我们将研究 在纤维化和改变的间充质成纤维细胞表型中的炎性浸润， 特异性抑制剂DCSL 1。我们将比较用DCSL-1处理的老年小鼠与年龄匹配的对照， 通过使用MRI的非侵入性细胞追踪研究浸润性白细胞，分析浸润性单核细胞， 淋巴细胞，并通过蛋白质阵列鉴定由它们分泌的细胞因子。而且 DCSL-1对T淋巴细胞群的作用将使用过继转移进行研究。对于体外研究， 还将分析分泌的细胞因子对小鼠和人来源的间充质成纤维细胞的作用。 最后，比较DCSL-1和对照治疗的老年小鼠的矩阵分析和心脏功能研究将在下文进行。 如SA 1中所示。这种方法是创新的，意义重大，因为它将使我们能够将心脏的变化联系起来。 分子和细胞反应的功能。此外，我们的初步数据显示， 小鼠模型与人类老化的心脏，在那里我们观察到同样的缺陷。