Animal and Clinical Core

摘要：通过使用胎儿心脏外科技术，Core C旨在提供PPG研究者 严格定义和可重复的先天性心脏病（CHD）新绵羊模型。为此目的， 核心C将全面生成，管理和提供所有与动物相关的实验，资源， 专业知识此外，核心C将从新生儿、婴儿、儿童和 患有各种不同先天性心脏病的成人，供PPG研究者使用。一个安全 已经建立了一个数据库，其中包含超过300名患者的库存样本和临床数据。在这一背景下， 核心C将通过提供六项服务来定义：1）CHD的绵羊模型。这将包括我们建立良好的 分流模型和我们新创建的LPA结扎模型。组织（外周肺和孤立血管）和 主肺动脉内皮细胞和平滑肌细胞（近端和微血管）将被 PPG研究者可用。2)数据的获取和分析。这将包括评价 心肺血流动力学、肺血管反应性（体内和体外）、 肺血管重建，先进的在体成像，体外循环对肺血管重建的影响 血液动力学和血管功能，以及不同疗法的效果。3)生化，增殖， 细胞凋亡和血管生成测定。这将包括各种活性氧的测量， 血管细胞和组织中的氧或氮氧化物的氮种类，细胞增殖，凋亡， 和管形成。4)不同生物力学力的体外评估。这包括利用 一种新的微流体腔室（UCSD），允许同时和独立的压力变化，循环 血管内皮细胞上的拉伸和流动诱导的剪切应力。5)CAR肽用于靶向 治疗受损的肺内皮细胞。为了减少脱靶效应的可能性，我们将利用 我们的疗法与CARSKNKDC（CAR）肽的IV共递送。6)生物储存库和相应的 新生儿、婴儿、儿童和成人CHD临床数据库。CHD患者的血液样本 将在手术修复之前和之后获得，用于人类确认最初鉴定的异常通路 用于动物和代谢筛查。确定的畸变与CHD类型之间的相关性（+/- 流量; +/-压力）以及术前和术后肺血管疾病的程度。 核心C将通过确保所有动物模型的标准化来加强所有三个项目的科学工作 并由经验丰富和训练有素的人员以统一的方式执行。调查人员曾有过 多年使用这些技术的经验运行Core C。他们开发或改编了几种 技术，并应用它们在体外和体内系统中进行生理学研究。作为中央 专用分析资源核心C将确保以具有成本效益的方式进行这些分析 动物发病率和死亡率最低。

ABSTRACT: With the use of fetal cardiac surgical techniques, Core C is designed to provide PPG investigators with rigorously defined and reproducible novel ovine models of congenital heart disease (CHD). To this end, Core C will comprehensively generate, manage and provide all animal-related experiments, resources, and expertise. In addition, Core C will obtain peri-operative blood samples from neonates, infants, children, and adults with a wide range of differing congenital heart defects, to be utilized by PPG investigators. A secure database is already established that contains banked samples and clinical data on >300 patients. In this context, Core C will be defined by providing six services: 1) Ovine models of CHD. This will include our well-established Shunt model and our newly created LPA ligation model. Tissues (peripheral lung and isolated vessels) and primary pulmonary artery endothelial and smooth muscle cells (both proximal and microvascular) will be available to PPG investigators. 2) Acquisition and analysis of data. This will include the evaluation of cardiopulmonary hemodynamics, pulmonary vascular reactivity (both in vivo and in vitro), the assessment of pulmonary vascular remodeling, advanced in vivo imaging, the effect of cardiopulmonary bypass on hemodynamics and vascular function, and the effect of different therapies. 3) Biochemical, proliferation, apoptosis, and angiogenesis determinations. This will include the measurement of various reactive oxygen and nitrogen species of oxygen or nitrogen oxide in vascular cells and tissues, indices of cell proliferation, apoptosis, and tube formation. 4) The in vitro assessment of differing biomechanical forces. This includes the utilization of a novel microfluidic chamber (UCSD) that allows the simultaneous and independent variation of pressure, cyclic stretch, and flow-induced shear stresses on the vascular endothelial cells. 5) Use of the CAR peptide to target therapeutics to the damaged pulmonary endothelium. To reduce the potential of off-target effects, we will utilize IV co-delivery of our therapies with the CARSKNKDC (CAR) peptide. 6) A bio-repository and corresponding clinical database of neonates, infants, children, and adults with CHD. Blood samples from patients with CHD will be obtained before and after surgical repair for human confirmation of aberrant pathways initially identified in animals and for metabolic screening. Correlation between the identified aberrations, with the type of CHD (+/- flow; +/- pressure), and degree of pre-operative and post-operative pulmonary vascular disease will be sought. Core C will enhance the scientific work for all three projects by assuring that all animal models are standardized and performed in a uniform manner by highly experienced and trained personnel. Investigators who have had many years of experience using these techniques run Core C. They have developed or adapted several of the techniques and applied them to perform physiological studies in in vitro and in vivo systems. As a central dedicated analytical resource Core C will ensure these analyses are also carried out in a cost-efficient manner with minimal animal morbidity and mortality.