Investigating the principles of physiological and pathological vascular remodeling via 4D imaging of live mouse skin

通过活体小鼠皮肤 4D 成像研究生理和病理血管重塑的原理

• 批准号: 10739431
• 负责人: Chen Yuan Kam
• 金额: $ 10.6万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10739431
• 关键词: 4D Imaging; Ablation; Address; Adult; Age; Architecture; Behavior; Birth; Blood Vessels; Blood flow; Cells; Cellular Morphology; Characteristics; Coupling; Cutaneous; Data; Development; Endothelial Cells; Endothelium; Epidermis; Genetic; Health; Homeostasis; Human; Image; Imaging Techniques; Imaging technology; Injectable; Injury; Kinetics; Lasers; Maintenance; Mediating; Membrane; Modeling; Molecular; Monitor; Morphology; Mus; Mutant Strains Mice; Natural regeneration; Neonatal; Nutrient; Organ; Pathologic; Pathology; Physiological; Play; Process; Property; Regenerative capacity; Regulation; Reporter; Research; Resolution; Role; Signal Transduction; Skin; Skin repair; Solid; Structure; Techniques; Technology; Temperature; Testing; Time; Tissues; VEGFA gene; Vascular Permeabilities; Vascular regeneration; Vascular remodeling; Vascularization; Visualization; cell behavior; cell motility; cell type; experimental study; hemodynamics; imaging approach; improved; in vivo; infancy; injury and repair; innovation; intravital imaging; migration; mouse model; multiphoton microscopy; mutant; neonate; network architecture; programs; regenerative; repaired; response; spatiotemporal; time use; tissue oxygenation; tool; transcriptomics; vascular abnormality; vessel regression; wound; wound healing; wound vascularization

PROJECT SUMMARY The cutaneous vasculature is a crucial yet understudied component of the skin, responsible for essential functions such as tissue oxygenation, exchange of nutrients and soluble factors, and temperature control. While we have a significant understanding of vascular abnormalities present in various skin pathologies, we lack an understanding of both physiological remodeling and homeostatic mechanisms sustaining lifelong function. Specifically, we lack the resolution of the coordinated cellular behaviors that drive developmental vascular remodeling, as well as those that underlie vascular regeneration in the face of injury, particularly in the context of hemodynamic status. I hypothesize that network-wide coordination of EC behaviors in relation to hemodynamic changes regulates developmental and regenerative remodeling programs of the skin vasculature. To test this hypothesis, I have established an intravital imaging technique that allows for the longitudinal tracking and manipulation of the endothelial cells (ECs) that constitute the lining of all blood vessels in the skin of a live mouse. In Aim 1, I will investigate the neonatal vessel remodeling program and underlying EC behaviors that orchestrate the establishment of skin vascular network architecture and blood flow efficiency. Following establishment of adult vascular homeostasis, I will probe the cellular mechanisms that regulate the maintenance of adult vessel integrity via a targeted laser ablation approach, modeling the discrete membrane damage inflicted upon the endothelium due to shear and contractile forces. My preliminary data shows that EC migration within existing vessel structures is a critical EC behavior that underlies network-wide vessel regression during neonatal remodeling, as well as the reparative response of adult ECs to local damage. In Aim 2, I will transition my studies towards the understanding of the skin vasculature in the context of pathological states. Firstly, I will investigate the wound vascularization mechanisms of neonatal versus adult skin, and delineate the differential remodeling properties that enable the enhanced wound revascularization in neonatal skin that we have observed in preliminary experiments. Second, I will determine how coordination of flow-dependent EC rearrangement impacts the ability of adult wounds to revascularize via a genetic mutant model that uncouples the ability of ECs to polarize with respect to blood flow direction. To achieve these aims, I will use an integrated approach of cutting-edge imaging technology, transcriptomics, and genetic mouse models. This research is significant because we expect to uncover global cellular and molecular mechanisms that coordinate vascular development, homeostasis, and injury repair. My findings will likely drive innovation in related fields, given the ubiquity and crucial roles of the vasculature in all organs.

项目概要 皮肤血管系统是皮肤的一个重要但尚未得到充分研究的组成部分，负责维持皮肤的基本功能 组织氧合、营养物质和可溶性因子的交换以及温度控制等功能。 虽然我们对各种皮肤病理中存在的血管异常有深入的了解，但我们 缺乏对维持终生的生理重塑和稳态机制的了解 功能。具体来说，我们缺乏驱动发育的协调细胞行为的解决方案 血管重塑，以及在受伤时血管再生的基础，特别是在 血流动力学状态的背景。我假设 EC 行为的网络范围协调 血流动力学变化调节皮肤的发育和再生重塑程序 脉管系统。为了检验这个假设，我建立了一种活体成像技术，可以 纵向追踪和操纵构成所有血液内层的内皮细胞 (EC) 活老鼠皮肤上的血管。 在目标 1 中，我将研究新生儿血管重塑计划和潜在的 EC 行为 协调皮肤血管网络结构的建立和血流效率。下列的 建立成人血管稳态，我将探讨调节血管稳态的细胞机制 通过有针对性的激光烧蚀方法维持成体血管的完整性，对离散膜进行建模 由于剪切力和收缩力对内皮造成的损伤。我的初步数据显示 EC 现有船舶结构内的迁移是网络范围船舶的关键 EC 行为 新生儿重塑过程中的回归，以及成人 EC 对局部损伤的修复反应。在 目标 2，我将把我的研究转向理解皮肤脉管系统 病理状态。首先，我将研究新生儿与成人的伤口血管化机制 皮肤，并描绘出不同的重塑特性，从而能够增强伤口血运重建 我们在初步实验中观察到的新生儿皮肤。其次，我将确定如何协调 血流依赖性 EC 重排通过基因突变影响成人伤口血运重建的能力 该模型解耦了 EC 相对于血流方向的极化能力。为了实现这些目标， 我将使用尖端成像技术、转录组学和基因小鼠的综合方法 模型。这项研究意义重大，因为我们期望揭示全球细胞和分子机制 协调血管发育、体内平衡和损伤修复。我的发现可能会推动创新 鉴于脉管系统在所有器官中的普遍性和关键作用，相关领域。