Determining the functions of LYVE1 on macrophages during autoantibody-induced valve disease

确定 LYVE1 在自身抗体诱导的瓣膜疾病期间对巨噬细胞的功能

• 批准号: 10534827
• 负责人: Victoria Osinski
• 金额: $ 6.76万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-01 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10534827
• 关键词: 3-Dimensional; Address; Adipose tissue; Arthritis; Attenuated; Autoantibodies; Automobile Driving; Binding; Bone Marrow; CD44 Antigens; Cardiovascular Diseases; Carditis; Cells; Characteristics; Chimera organism; Coculture Techniques; Coupled; Data; Disease; Disease Progression; ECM receptor; Endothelium; Environment; Extracellular Matrix; Fellowship; Flow Cytometry; Frequencies; Gene Expression; Gene Expression Profile; Genes; Growth; Heart; Heart Atrium; Heart Diseases; Heart Valves; Histology; Homeostasis; Homing; Hyaluronan; Image; In Vitro; Inflammation; Inflammatory; Innate Immune Response; Knockout Mice; Link; Location; LoxP-flanked allele; Lung; Lymphangiogenesis; Lymphatic; Lymphatic Endothelial Cells; Measures; Mediating; Mitral Valve; Modeling; Mus; Peritoneum; Population; Population Heterogeneity; Production; Proteins; Publishing; Pulmonary Fibrosis; Reporter; Reporting; Research; Resources; Rheumatic Heart Disease; Rheumatism; Rheumatoid Arthritis; Role; Science; Structure; Systemic Lupus Erythematosus; Techniques; Testing; Thick; Time; Tissue imaging; Tissues; Transcript; Work; angiogenesis; cardiovascular disorder risk; density; diagnostic biomarker; experience; heart function; human disease; immune activation; improved; in vivo; lymphatic valve; macrophage; mouse model; neovascular; new therapeutic target; novel marker; pre-clinical; single cell sequencing; tool; transcriptome sequencing

PROJECT SUMMARY/ABSTRACT Rheumatic diseases including rheumatoid arthritis, rheumatic heart disease, and systemic lupus erythematosus are associated with increased risk of cardiovascular disease. While inflammation is a critical link between rheumatic diseases and CVD, continued research is needed to determine causal mechanisms. The K/B.g7 model of spontaneous autoantibody-induced arthritis and mitral valve (MV) disease provides a valuable pre- clinical tool to elucidate disease-driving mechanisms related to valvular carditis, which is currently difficult to detect prior to the onset of reduced cardiac function. Prior work from our group demonstrated a role for macrophages in promoting valve disease. Single-cell sequencing data from inflamed MV reported a heterogeneous population of macrophages in this tissue prompting further inquiry into the macrophage subsets and subset-specific roles that drive disease. Preliminary data from our group demonstrated that lymphatic endothelial cells promote early stages of MV disease, but the mechanisms driving the growth of these neo- lymphatic structures remains unclear. We identified the presence of hyaluronan receptor LYVE1+ macrophages near the atrial layer of endothelium and MV LECs in both normal and diseased valves. Gene expression and flow cytometry profiling revealed that these cells are tissue resident macrophages. LYVE1+ macrophages have been identified in other locations in the heart as well as other tissues including the lung, adipose, and peritoneum. In these niches, LYVE1+ macrophages interact with tissue native hyaluronan, regulate extracellular matrix homeostasis, and promotes neovascular growth. Work proposed in this fellowship will test the hypothesis that a) early in disease, LYVE1+ macrophages promote MV disease at least in part by promoting lymphangiogenesis and b) the LYVE1 protein does not simply delineate this population of macrophages but is required for the macrophages to perform these functions. Using RNA sequencing, LYVE1+ and LYVE1- MV macrophage subsets will be compared to one another and across disease time points to identify the gene expression profile unique to LYVE1+ macrophages, which we predict will include lymphangiogenesis- and HA-binding-related transcripts. Beyond characterizing these cells, an inducible model of LYVE1+ macrophage depletion and LYVE1 WT and KO bone marrow chimeras will be used to test the necessity of this cell population and LYVE1 expression on macrophages, respectively, in driving both MV disease and lymphatic growth in diseased valves. Deep-tissue imaging and analysis techniques will be applied to further interrogate whether LYVE1+ macrophages preferentially interact with LECs to promote vessel growth. In all, these studies will elucidate important information about the role of innate immune responses in driving valvular carditis. These data can be used to improve our understanding of the early mechanisms driving human disease and potentially identify novel therapeutic targets or diagnostic markers.

项目概要/摘要 风湿性疾病，包括类风湿性关节炎、风湿性心脏病和系统性红斑狼疮 与心血管疾病风险增加有关。虽然炎症是两者之间的关键联系 风湿性疾病和心血管疾病，需要继续研究以确定因果机制。 K/B.g7 自发性自身抗体诱导的关节炎和二尖瓣（MV）疾病模型提供了有价值的预 阐明与瓣膜性心脏炎相关的疾病驱动机制的临床工具，目前很难 在心功能下降之前进行检测。我们小组之前的工作证明了 巨噬细胞促进瓣膜疾病。来自发炎 MV 的单细胞测序数据报告了 该组织中巨噬细胞的异质群体促使进一步研究巨噬细胞亚群 以及驱动疾病的子集特定作用。我们小组的初步数据表明，淋巴管 内皮细胞促进 MV 疾病的早期阶段，但驱动这些新细胞生长的机制 淋巴结构仍不清楚。我们确定了透明质酸受体 LYVE1+ 巨噬细胞的存在 正常和患病瓣膜中的内皮和 MV LEC 的心房层附近。基因表达和 流式细胞术分析显示这些细胞是组织驻留巨噬细胞。 LYVE1+ 巨噬细胞具有 已在心脏的其他部位以及肺、脂肪和腹膜等其他组织中发现。 在这些生态位中，LYVE1+巨噬细胞与组织天然透明质酸相互作用，调节细胞外基质 体内平衡，促进新血管生长。本研究金中提出的工作将检验以下假设： a) 在疾病早期，LYVE1+巨噬细胞至少部分通过促进淋巴管生成来促进 MV 疾病 b) LYVE1 蛋白不仅仅描绘了巨噬细胞群，而且是 巨噬细胞执行这些功能。使用 RNA 测序，LYVE1+ 和 LYVE1- MV 巨噬细胞 子集将在疾病时间点之间相互比较，以确定基因表达谱 LYVE1+巨噬细胞所特有的，我们预测其将包括与淋巴管生成和HA结合相关的 成绩单。除了表征这些细胞之外，LYVE1+ 巨噬细胞耗竭和 LYVE1 的诱导模型 WT和KO骨髓嵌合体将用于测试该细胞群和LYVE1表达的必要性 分别作用于巨噬细胞，驱动MV疾病和病变瓣膜中的淋巴生长。深层组织 成像和分析技术将用于进一步询问 LYVE1+ 巨噬细胞是否 优先与 LEC 相互作用以促进血管生长。总而言之，这些研究将阐明重要的 有关先天免疫反应在驱动瓣膜性心脏炎中的作用的信息。这些数据可用于 提高我们对驱动人类疾病的早期机制的理解，并有可能发现新的机制 治疗靶点或诊断标记。