ENDOTHELIAL CELL GROWTH AND MATURATION IN A 3D CULTURE

研究：原发性肺动脉高压(PPH)的肺动脉结构改变以内侧肌肉肥厚和称为丛状病变的复杂血管病变为特征。应用内皮细胞(EC)特异性标记物对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，结果显示，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。这项建议的总体目标是检查切应力和血管生成调节剂对EC生长和功能的影响，以此作为了解与丛状病变相关的PH发病机制的一种手段。我们假设剪切力导致血管内皮生长因子和/或血管内皮生长因子受体表达增加，导致内皮细胞功能障碍和增殖。由于血管生成素(Angiopoietin，Ang)1、Ang2和VEGF可促进血管内皮细胞的逐渐成熟和组织化，我们还假设血管生成配体(VEGF、Ang1、Ang2)与它们的酪氨酸激酶受体相互作用，以维持胚胎血管生成表型的生长。为了研究内皮细胞在PPH丛状病变发生中的作用的机制问题，我们将使用三维、微血管培养系统对体外血管生成进行系统的研究。慢性血流系统的初步工作表明，剪切力、三维、体外ECs和剪切力条件之间的形态相似，将使用重组生长因子或酪氨酸激酶调节、增殖和生长的抑制剂来分析非PPG条件下的生长，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的整个过程中继续进行。在这一提议期间，我将获得思想和调查方法的独立性，以过渡到独立的研究人员地位，对实验性肺部病理学领域有兴趣和专业知识。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.