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.

项目摘要 皮肤脉管系统是皮肤的重要但未充分研究的组成部分，负责皮肤的基本功能。 功能如组织氧合、营养物和可溶性因子的交换以及温度控制。 虽然我们对各种皮肤病变中存在的血管异常有了很深的了解， 缺乏对维持终身的生理重塑和自我平衡机制的理解， 功能具体来说，我们缺乏对协调细胞行为的解决方案，这些行为推动了发育 血管重塑，以及那些在面对损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，网络范围内的电子商务行为的协调， 调节皮肤的发育和再生重塑程序 脉管系统为了验证这一假设，我建立了一种活体成像技术， 纵向跟踪和操纵构成所有血液衬里的内皮细胞（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和血流效率的建立。以下 为了建立成人血管内稳态，我将探讨调节血管内稳态的细胞机制。 通过靶向激光消融方法维持成人血管完整性，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。初步数据显示， 在现有血管结构内的迁移是一种关键的EC行为，它是网络范围血管的基础 新生儿重塑过程中的退化，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对皮肤血管系统的理解， 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，并描绘差异重塑特性，使增强伤口血运重建， 我们在初步实验中观察到的新生儿皮肤。第二，如何协调 血流依赖性EC重排通过基因突变影响成人伤口再血管化的能力 该模型将EC的能力与血流方向分离。为了实现这些目标， 我将使用尖端成像技术，转录组学和遗传小鼠的综合方法 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 鉴于脉管系统在所有器官中的普遍性和关键作用，