ENDOTHELIAL CELL GROWTH AND MATURATION IN A 3D CULTURE

研究：原发性肺动脉高压（PPH）的肺动脉结构改变的特征是内侧肌肉增厚和复杂的血管病变，称为丛状病变。利用内皮细胞（EC）特异性标志物对PPH和继发性肺动脉高压（PH）患者的丛状病变进行三维计算机重建显示，丛状病变的肺内皮细胞处于不同的发育阶段，其增殖明显阻碍肺血流。本提案的总体目标是研究剪切应力和血管生成调节剂对EC生长和功能的影响，作为了解与丛状病变相关的PH发病机制的一种手段。我们假设剪切应力导致VEGF和/或VEGF受体表达增加，导致内皮细胞功能障碍和增殖。由于血管生成素（Ang1）、Ang2和VEGF导致血管腔内内皮细胞的逐渐成熟和组织，我们也假设血管生成配体（VEGF、Ang1、Ang2）相互作用并与其酪氨酸激酶受体相互作用以维持胚胎血管生成表型生长。为了研究内皮细胞在PPH丛状病变发生中的作用机制问题，我们将使用三维微血管内皮细胞培养系统进行体外血管生成的系统研究。对慢性血流系统的初步研究表明，剪切应力、三维体外ECs和剪切应力条件下的ECs在形态上存在相似性。研究人员将利用重组生长因子或酪氨酸激酶抑制剂来分析非ppg条件下的增殖和生长，如肿瘤血管生成、伤口修复和愈合、女性生殖和糖尿病视网膜病变。候选人和环境：Rubin Tuder博士实验室的正式指导将在本提案期间持续进行。在此提案期间，我将获得独立思考和研究方法的独立性，以过渡到独立的资助研究人员地位，并对实验性肺病理学领域感兴趣和专业知识。一个咨询委员会已经成立，以帮助我的学术生涯发展。

Research: Structural alterations of the pulmonary arteries in primary pulmonary hypertension (PPH) are characterized by medial muscular thickening and complex vascular lesions known as plexiform lesions. Three-dimensional computerized reconstruction of plexiform lesions from PPH and secondary pulmonary hypertension (PH) patients using endothelial cell (EC) specific markers, shows that the pulmonary ECs of plexiform lesions are in different developmental stages and that their proliferation markedly impedes pulmonary blood flow. The overall goal of this proposal is to examine the effects of shear stress and angiogenic regulators on EC growth and function-as a means to understand the pathogenesis of PH associated with plexiform lesions. We hypothesize that shear stress causes increased VEGF and/or VEGF receptor expression, leading to endothelial cell dysfunction and proliferation. Since Angiopoietin (Ang)1, Ang2, and VEGF cause progressive maturation and organization of ECs lining blood vessel lumina, we also hypothesize that the angiogenic ligands (VEGF, Ang1, Ang2) interact with each other and their tyrosine kinase receptors to maintain an embryonic vasculogenic phenotypic growth. To study the mechanistic questions related to the role of ECs in the genesis of plexiform lesions in PPH, we will use a three-dimensional, microvascular EC culture system for systematic studies of angiogenesis in vitro. Preliminary work with the chronic flow system demonstrates a morphologic similarity between the shear-stressed, three-dimensional, in vitro ECs and shear stress conditions will be analyzed with the use of recombinant growth factors or inhibitors of the tyrosine kinase regulation, proliferation, and growth in non-PPG conditions, such as tumor angiogenesis, wound repair and healing , female reproduction, and diabetic retinopathy. Candidate and Environment: Formal mentorship in the lab of Dr. Rubin Tuder will continue throughout the time of this proposal. During this proposal period, I will acquire the independence in though and investigative methods required for the transitions to independent status as a funded researcher, with interest and expertise in the area of experimental pulmonary pathology. An Advisory Committee has been established to aid my academic career development.