Animal and Clinical Core

摘要：通过使用胎儿心脏外科技术，Core C旨在为PPG调查人员提供 具有严格定义和可重复性的先天性心脏病(CHD)的新型绵羊模型。为此， 核心C将全面生成、管理和提供所有与动物相关的实验、资源和 专业知识。此外，Core C将获取新生儿、婴儿、儿童和 患有各种不同先天性心脏病的成年人，供PPG调查人员使用。一个安全的地方 数据库已经建立，其中包含储存的样本和300名患者的临床数据。在这方面， 核心C将通过提供六项服务来定义：1)CHD的绵羊模型。这包括我们久负盛名的 分流模型和我们新建立的LPA结扎模型。组织(外周肺和隔离血管)和 原代肺动脉内皮细胞和平滑肌细胞(包括近端和微血管)将 可供PPG调查人员使用。2)数据的采集与分析。这将包括对 心肺血流动力学，肺血管反应性(体内和体外)，评估 肺血管重塑，先进的活体成像，体外循环对 血流动力学和血管功能，以及不同治疗方法的效果。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和 血管细胞和组织中氧或氮氧化物的氮物种，细胞增殖，细胞凋亡， 和管子的形成。4)不同生物力学力的体外评价。这包括利用 一种新型的微流体室(UCSD)，允许同时和独立地改变压力，循环 血管内皮细胞的拉伸和流动引起的剪应力。5)使用CAR多肽作为靶点 肺内皮损伤的治疗。为了减少偏离目标的潜在影响，我们将利用 IV与CARSKNKDC(CAR)肽联合提供我们的治疗。6)生物信息库和相应的 新生儿、婴儿、儿童和成人冠心病临床数据库。冠心病患者的血液样本 将在手术修复前后获得，以供人类确认最初识别的异常通路 用于动物和新陈代谢筛查。识别的像差与CHD类型(+/-)之间的相关性 血流；+/-压力)，以及手术前后肺血管病变的程度。 核心C将通过确保所有动物模型标准化来加强所有三个项目的科学工作 并由经验丰富、训练有素的人员统一执行。调查人员有过 多年使用这些技术的经验运行Core C。他们已经开发或改编了几个 技术，并将其应用于体外和体内系统的生理学研究。作为一个中心 专门的分析资源核心C将确保这些分析也以经济高效的方式进行 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.