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: 10611152
• 负责人: KATARZYNA A. CIESLIK
• 金额: $ 16万
• 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 2023-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10611152
• 关键词: Activation Analysis; Adoptive Transfer; Age; Aging; Binding; CD209 gene; Cardiac Function Study; Cells; Collagen; Cytometry; Data; Defect; Deposition; Development; Elderly; Electron Microscopy; Epidemic; Fibroblasts; Fibronectins; Fibrosis; Functional disorder; Heart; Human; In Vitro; Inflammatory; Inflammatory Infiltrate; Leukocytes; Link; Lymphocyte; Magnetic Resonance Imaging; Mass Spectrum Analysis; Measurement; Mesenchymal; Mission; Molecular; Mus; Myelogenous; Nature; Pathologic; Pathway interactions; Persons; Phenotype; Population; Process; Protein Array; Public Health; Research; Role; Signal Pathway; Signal Transduction; T-Lymphocyte; Work; aged; aging population; base; coronary fibrosis; cytokine; heart function; human tissue; improved; in vivo; inhibitor; innovation; interstitial; macrophage; monocyte; mouse model; non-invasive monitor; receptor; response

PROJECT SUMMARY 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.

项目摘要 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纤维化 在老化的心脏。中心假设来自我们的数据，我们在这些数据中发现了信号异常， 在间充质成纤维细胞中，其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。单核细胞浸润到骨髓成纤维细胞中，进一步促进 间充质成纤维细胞病理活化。检查每种成纤维细胞类型及其作用 在这种相互作用中，我们将使用特异性抑制剂，例如1）AICAR，一种AMPK激活剂，其在病理学上抑制 上调间充质成纤维细胞中的Erk途径，从而降低间充质成纤维细胞中的胶原蛋白和纤连蛋白水平。 2）DCSL-1，其特异性结合DC-SIGN，DC-SIGN是一种仅在白细胞中发现的受体。我们 已经发现体内DCSL-1治疗改善心脏功能并减少M1的数量， 心脏中的促炎性巨噬细胞和骨髓成纤维细胞以及减少胶原沉积。在 SA 1，我们将确定缺陷的间充质成纤维细胞促进间质细胞增殖的机制。 研究衰老心脏中的纤维化，并检查AMPK激活剂在逆转这些缺陷中的作用。我们将研究 病理性基质沉积在老年小鼠和人体组织中发生的分子机制 使用电子显微镜、质谱和信号通路活化分析。我们还将 确定体内和体外AICAR对这些过程的影响。最后，心脏纤维化的变化 在小鼠体内AICAR治疗后，将通过非侵入性MRI、Echo和Doppler监测功能 测量.在SA 2中，我们将研究炎性浸润在纤维化中的作用，以及在改变的细胞中的作用。 间充质成纤维细胞表型通过使用骨髓特异性抑制剂，DCSL 1。我们将老年小鼠 用DCSL-1与年龄匹配的对照治疗，并通过非侵入性细胞追踪研究浸润性白细胞 使用MRI，通过质谱细胞术分析浸润的单核细胞和淋巴细胞，并鉴定分泌的细胞因子 通过蛋白质阵列。此外，将研究DCSL-1对T淋巴细胞群的作用 使用过继转移。用于体外研究分泌的细胞因子对小鼠间充质成纤维细胞的作用 人类起源也将被分析。最后通过矩阵分析和心功能研究比较DCSL- 1和对照处理的老年小鼠将如SA 1中那样进行。这种方法具有创新性和重要性，因为它 将使我们能够将心脏功能的变化与分子和细胞反应联系起来。此外，我们的初步 数据显示了小鼠模型与人类衰老心脏的相关性，我们观察到了相同的缺陷。

项目成果

Mechanosensing dysregulation in the fibroblast: A hallmark of the aging heart.

• DOI: 10.1016/j.arr.2020.101150
• 发表时间: 2020-11
• 期刊: Ageing research reviews
• 影响因子: 13.1
• 作者: Angelini A;Trial J;Ortiz-Urbina J;Cieslik KA
• 通讯作者: Cieslik KA

Aortic acceleration as a noninvasive index of left ventricular contractility in the mouse.

• DOI: 10.1038/s41598-020-79866-y
• 发表时间: 2021-01-12
• 期刊: Scientific reports
• 影响因子: 4.6
• 作者: Perez JET;Ortiz-Urbina J;Heredia CP;Pham TT;Madala S;Hartley CJ;Entman ML;Taffet GE;Reddy AK
• 通讯作者: Reddy AK

Treatment with a DC-SIGN ligand reduces macrophage polarization and diastolic dysfunction in the aging female but not male mouse hearts.

• DOI: 10.1007/s11357-020-00255-4
• 发表时间: 2021-04
• 期刊: GeroScience
• 影响因子: 5.6
• 作者: Trial J;Diaz Lankenau R;Angelini A;Tovar Perez JE;Taffet GE;Entman ML;Cieslik KA
• 通讯作者: Cieslik KA