Integration of advanced imaging and multiOMICs to elucidate pro-atherogenic effects of endothelial-to-Immune cell-like transition (EndICLT)

整合先进成像和多组学技术来阐明内皮细胞向免疫细胞样转变的促动脉粥样硬化效应 (EndICLT)

• 批准号: 10606258
• 负责人: Kyung In Baek
• 金额: $ 7.43万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-02-01 至 2024-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10606258
• 关键词: 3-Dimensional; Accounting; Address; Antiatherogenic; Arterial Fatty Streak; Arteries; Atherosclerosis; Atlases; Attenuated; Biochemical Reaction; Biological; Biological Markers; Blood Viscosity; Blood flow; CD44 gene; CDH5 gene; Cardiovascular system; Cause of Death; Cell Reprogramming; Cells; Cellular Assay; Chromatin; Chronic; Common carotid artery; Complementary RNA; Confocal Microscopy; Cues; Data; Detection; Disease; Endothelial Cells; Endothelium; Environment; Fluorescence; Fluorescence Microscopy; Fluorescent in Situ Hybridization; Foundations; Genes; Genetic Transcription; Goals; Grant; Hydrogels; Image; Immune; Inflammation; Inflammatory Response; Isotopes; Label; Lateral; Left; Ligation; Light; Lighting; Link; Machine Learning; Macrophage; Maintenance; Maps; Medial; Mediating; Mesenchymal; Methods; Methylene Chloride; Modality; Modeling; Morbidity - disease rate; Morphology; Mus; Nature; Noise; Nucleic Acids; Ontology; Optics; PECAM1 gene; Pathway interactions; Penetration; Periodicity; Pharmaceutical Preparations; Phenotype; Procedures; Process; Proliferating; Proteins; Qualitative Evaluations; Quantitative Evaluations; RNA; RNA Sequences; Reaction; Research; Resolution; Role; Signal Transduction; Smooth Muscle Myocytes; Specimen; Stents; Structure; Techniques; Testing; Therapeutic; Time; Tissues; Touch sensation; Transposase; VWF gene; Vascular Cell Adhesion Molecule-1; Visualization; atherogenesis; career; cell type; chromophore; endothelial stem cell; epigenomics; experience; fluorescence imaging; genome-wide; hypercholesterolemia; imaging Segmentation; imaging approach; imaging platform; in vivo; insight; mortality; mouse model; multidisciplinary; multiple omics; new therapeutic target; novel; novel strategies; preservation; prevent; reconstruction; recruit; shear stress; single-cell RNA sequencing; spatiotemporal; systemic inflammatory response; therapeutic target

Project Summary/Abstract Atherosclerosis is a multifactorial disease accounting for a leading cause of morbidity and mortality. The endothelium, the inner lining of vessel walls, transduces constant and rhythmic wall shear stress (WSS) from blood viscosity and flow. At the medial wall of arterial bifurcation, stable unidirectional laminar flow (S-flow) develops attenuates systemic inflammatory responses, whereas bidirectional and axially misaligned flow in the lateral wall determine focal but eccentric nature of chronic low-grade inflammatory responses, endothelial cell reprogramming and preferential formation of atherosclerotic lesion. Our unbiased scRNA seq and gene ontology analysis suggested that D-flow induces endothelial-to-immune cell like-transition (EndICLT) and pro-atherogenic pathways. However, the pathophysiological significance of EndICLT in vivo and whether it could serve as an anti-atherogenic therapeutic target is yet remains elusive. One of the longstanding technical challenges is interrogating cell signaling machinery and function simultaneously to unravel key mechanisms in action and dynamic changes in pathophysiological milestones. Comprehensive insights into atherosclerosis and endothelial dynamics can be achieved by visualizing multiomic atlas throughout the plaque. Cutting-edge fluorescence microscopy in optically cleared plaque provides qualitative and quantitative evaluations of atherosclerosis. However, current modalities of fluorescence imaging and time demanding procedures of conventional tissue clearing techniques limits high throughput imaging with high spatial resolution. The advent of multi-scale sub- voxel light-sheet fluorescence microscopy combined with a rapid clearing of plaques may address unmet challenges. In this grant, we will use novel approaches of tissue clearing, advanced image acquisition to elucidate flow-sensitive mechanisms, whereby EndICLT promotes atherosclerosis. In Aim1, we will examine whether the novel dichloromethane - histodenz gradient medium clearing technique has potential for rapid extraction of multiomic information in optically cleared plaque. Conventional fluorescence microscopy techniques including wide-field and diffraction-limited confocal microscopy creates interference from out-of-focus illumination and reduced axial penetration depth across the specimen. In Aim 2, we will focus on establishing an advanced imaging platform of LSFM followed by sub-voxel reconstruction (SV-LSFM) and machine-learning based image segmentation for scalable extraction of multiomic features in high spatial resolution. Finally, in Aim 3, we will integrate Aim 1 and 2 to explore key underlying mechanisms in the EndICLT-dependent atherogenesis. Together, these aims will allow a paradigm shift to identify novel therapeutic targets of atherosclerosis. This proposal will allow me to deepen my research experiences and provide a critical support to build a strong foundation for a career in cardiovascular research.

项目摘要/摘要 动脉粥样硬化是一种多因素疾病，占发病率和死亡率的主要原因。这 内皮是容器壁的内壁，从 血粘度和流动。在动脉分叉的内侧壁，稳定的单向层流（S-Flow） 发展会减弱系统性炎症反应，而双向和轴向不一致的流动 侧壁确定慢性低度炎症反应的局灶性但偏心的性质，内皮细胞 动脉粥样硬化病变的重编程和优先形成。我们公正的SCRNA SEQ和基因本体论 分析表明，D-Flow诱导内皮到免疫细胞类似转变（Endiclt）和亲动脉粥样硬化 途径。但是，始于体内的恩迪克特的病理生理意义，以及它是否可以作为一个 抗动脉生就治疗靶点仍然难以捉摸。长期以来的技术挑战之一是 询问细胞信号机械和功能，以揭示行动中的关键机制 病理生理里程碑的动态变化。对动脉粥样硬化和内皮的全面见解 可以通过在整个牌匾中可视化多瘤地图集来实现动力学。尖端的荧光 光学清除牙菌斑中的显微镜提供了对动脉粥样硬化的定性和定量评估。 但是，当前荧光成像和常规组织需要时间要求的方法 清除技术限制了高空间分辨率的高吞吐量成像。多尺度子的出现 体素灯页荧光显微镜与快速清理斑块结合了可能未满足的斑块 挑战。在这笔赠款中，我们将使用新颖的组织清除方法，高级图像获取来阐明 流动敏感机制，从而促进动脉粥样硬化。在AIM1中，我们将检查是否 新型二氯甲烷 - Histodenz梯度培养基清除技术有可能快速提取 光学清除牌匾中的多构信息。传统的荧光显微镜技术包括 宽场和衍射受限的共聚焦显微镜会因异常照明而产生干扰 轴向穿透深度减少了整个样品。在AIM 2中，我们将专注于建立高级 LSFM的成像平台，然后是亚素重建（SV-LSFM）和基于机器学习的图像 在高空间分辨率中可扩展多素特征的分割。最后，在AIM 3中，我们将 集成目标1和2，以探索依赖于依赖性的动脉粥样硬化中的钥匙潜在机制。一起， 这些目标将允许范式转移以识别动脉粥样硬化的新治疗靶标。该提议将 允许我加深我的研究经验，并提供关键的支持，以建立强大的基础 心血管研究的职业。