Temporospatial Single-Cell Characterization of Angiogenesis and Myocardial Regeneration in Small and Large Mammals

小型和大型哺乳动物血管生成和心肌再生的时空单细胞表征

• 批准号: 10751870
• 负责人: Stefan Elde
• 金额: $ 7.61万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-10 至 2025-08-09
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10751870
• 关键词: Acute; Adult; Advisory Committees; Affect; Aftercare; Angiogenesis Modulating Agents; Animal Model; Antigens; Architecture; Arteries; Award; Bar Codes; Blood Vessels; Bone Marrow; CXCR4 Receptors; Cardiac Myocytes; Cell physiology; Cells; Complex; Coronary Artery Bypass; Cues; DNA; Data; Detection; Endothelial Cells; Extracellular Matrix; Family suidae; Fellowship; Funding; Future; Goals; Grant; Heart; Heart failure; Human; Immunofluorescence Microscopy; In Situ; Infarction; Inflammatory; Inflammatory Response; Injury; K-Series Research Career Programs; Lead; Life; Ligation; Link; Macrophage; Mammals; Mediator; Mentorship; Modeling; Morbidity - disease rate; Mus; Myocardial; Myocardial Infarction; Myocardial Ischemia; Myocardium; Natural regeneration; Neighborhoods; Neonatal; Newborn Infant; Paracrine Communication; Pathologic; Pathway interactions; Patients; Perfusion; Persons; Phenotype; Physicians; Positioning Attribute; Postdoctoral Fellow; Process; Proliferating; Publishing; Recovery; Regenerative pathway; Regulatory Pathway; Research Training; Residual state; Resolution; Role; Science; Scientist; Signal Pathway; Signal Transduction; Stromal Cell-Derived Factor 1; Surgeon; Techniques; Therapeutic; Time; Tissues; Translational Research; United States National Institutes of Health; Vascularization; Ventricular Remodeling; Visualization; Work; angiogenesis; antibody conjugate; cell motility; cell type; chemokine; clinically significant; imaging facilities; imaging platform; indexing; injured; insight; interest; ischemic injury; migration; monocyte; mortality; mouse model; multidisciplinary; multiparametric imaging; neonatal mice; neonatal period; neonate; neovascularization; neovasculature; new therapeutic target; novel; paracrine; percutaneous coronary intervention; porcine model; programs; recruit; regeneration potential; regenerative; repaired; single nucleus RNA-sequencing; spatial relationship; spatiotemporal; wound healing

Project Summary/Abstract Ischemic heart disease affects more than 197 people worldwide every year. Despite advances in macro revascularization techniques such as coronary artery bypass grafting and percutaneous coronary intervention, many patients progress to heart failure due to residual microvascular perfusion deficits. Although the adult human heart appears to be unable to significantly regenerate myocardium after injury, neonatal mice and pigs are capable of efficient angiogenesis and myocardial regeneration during the first week of life. Endogenous angiogenic and regenerative pathways are intricately linked to the wound-healing inflammatory cascade, extracellular matrix remodeling, endothelial cell migration, and cardiomyocyte proliferation. Consequently, the infarct border zone, which is the spatial intersection of these cellular processes, has proven to be a complex and dynamic microenvironment to investigate. In this proposal, we describe a novel application of a multiplexed immunofluorescent imaging platform with single cell resolution called CODEX (co-detection by indexing) which was developed here at Stanford. With the guidance and mentorship described in the research training plan, we have compiled an advisory committee of physician and surgeon-scientists, cardiologists, cell biologists, and the experts in CODEX from Stanford's Cell Sciences Imaging Facility. If awarded this Fellowship, this multidisciplinary team is uniquely positioned to execute this first of its kind application of CODEX to myocardial regeneration. CODEX will be used to characterize the longitudinal changes in the spatiotemporal relationship between the resident cells of the myocardium and the paracrine pathways that govern angiogenesis and myocardial regeneration. CODEX is an extension of immunofluorescence microscopy that utilizes antibodies conjugated to DNA barcodes to simultaneously quantify up to 44 antigens in situ. In Aim 1 we will apply CODEX to neonatal mouse and pig LAD ligation models of myocardial regeneration to define novel spatial phenotypes of angiogenic and inflammatory cellular neighborhoods throughout the neonatal period of regenerative potential in comparison to non-regenerating adults. After characterizing the cellular microenvironments and cell-signaling activity of natural angiogenesis in neonatal mammals, in Aim 2 we will investigate the effect of exogenous modulators of angiogenesis and the acute inflammatory response in adult mice and pigs. As an animal model for myocardial regeneration, the regulatory pathways that govern natural angiogenesis and subsequent myocardial regeneration in the mouse and pig are the subject of great interest due to their potential therapeutic benefit in humans. By using CODEX to study the complex interplay between multiple cell types, paracrine signaling pathways, and intercellular processes we hope to advance our mechanistic understanding of natural neonatal angiogenesis and myocardial regeneration and identify new therapeutic targets for ischemic heart disease, which has the potential to advance the treatment of millions of patients worldwide who are suffering from ischemic heart disease.

项目概要/摘要 缺血性心脏病每年影响全球超过 197 人。尽管宏观方面取得了进步 血运重建技术，例如冠状动脉旁路移植术和经皮冠状动脉介入治疗， 许多患者由于残余微血管灌注不足而进展为心力衰竭。虽然成年人类 心脏受伤后似乎无法显着再生心肌，新生小鼠和猪 能够在生命的第一周内有效地进行血管生成和心肌再生。内源性 血管生成和再生途径与伤口愈合炎症级联密切相关， 细胞外基质重塑、内皮细胞迁移和心肌细胞增殖。因此， 梗死边界区是这些细胞过程的空间交叉点，已被证明是一个复杂且复杂的区域。 动态微环境进行研究。在这个提案中，我们描述了一种多路复用的新颖应用 具有单细胞分辨率的免疫荧光成像平台，称为 CODEX（索引联合检测）， 是在斯坦福大学开发的。在研究培训计划中描述的指导和指导下，我们 组建了一个由内科医生和外科医生科学家、心脏病专家、细胞生物学家和 来自斯坦福大学细胞科学成像中心的 CODEX 专家。如果获得此奖学金，则 多学科团队处于独特的地位，可以执行 CODEX 在心肌方面的首次应用 再生。 CODEX将用于表征时空关系的纵向变化 心肌驻留细胞和控制血管生成的旁分泌途径之间 心肌再生。 CODEX 是利用抗体的免疫荧光显微镜的延伸 与 DNA 条形码缀合，可同时原位定量多达 44 种抗原。在目标 1 中，我们将应用 CODEX 新生小鼠和猪心肌再生 LAD 结扎模型以确定新的空间表型 整个新生儿期的血管生成和炎症细胞邻域的再生潜力 与非再生成人相比。在表征细胞微环境和细胞信号传导后 新生哺乳动物中自然血管生成的活性，在目标 2 中，我们将研究外源性血管生成的影响 成年小鼠和猪的血管生成和急性炎症反应的调节剂。作为动物模型 对于心肌再生，控制自然血管生成和随后的调节途径 小鼠和猪的心肌再生因其潜在的治疗作用而受到极大关注 对人类有益。通过使用 CODEX 研究多种细胞类型之间复杂的相互作用，旁分泌 信号通路和细胞间过程，我们希望增进我们对自然机制的理解 新生儿血管生成和心肌再生并确定缺血性心脏的新治疗靶点 疾病，有可能促进全世界数百万患有该病的患者的治疗 来自缺血性心脏病。