ENDOTHELIAL CELL GROWTH AND MATURATION IN A 3D CULTURE

3D 培养中的内皮细胞生长和成熟

• 批准号: 6536532
• 负责人: CARLYNE D COOL
• 金额: $ 12.31万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-07-01 至 2004-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6536532
• 关键词: angiogenesis; angiogenesis factor; cell differentiation; cell growth regulation; cell proliferation; electron microscopy; embryo /fetus tissue /cell culture; enzyme linked immunosorbent assay; growth factor receptors; human tissue; immunocytochemistry; in situ hybridization; mechanical stress; phenotype; protein tyrosine kinase; pulmonary artery; pulmonary hypertension; receptor expression; shear stress; tissue /cell culture; transfection; vascular endothelial growth factors; vascular endothelium

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.

调研：原发性肺动脉高压（PPH）肺动脉结构改变的特征是中膜肌增厚和复杂的血管病变（称为丛状病变）。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，显示丛状病变的肺EC处于不同的发育阶段，其增殖明显阻碍肺血流。这个建议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，剪切应力引起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（Ang）1，Ang 2和VEGF导致内皮细胞内衬血管腔的逐步成熟和组织化，我们还假设血管生成配体（VEGF，Ang 1，Ang 2）相互作用和酪氨酸激酶受体，以维持胚胎血管生成表型生长。为了研究与PPH中丛状病变发生中EC作用相关的机制问题，我们将使用三维微血管EC培养系统进行体外血管生成的系统研究。慢性血流系统的初步研究表明，剪切应力、三维、体外EC和剪切应力条件之间的形态学相似性将通过使用重组生长因子或酪氨酸激酶调节、增殖和生长抑制剂在非PPG条件下（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）进行分析。候选人和环境：鲁宾·图德博士实验室的正式指导将在本提案的整个过程中继续。在此建议期间，我将获得独立的思想和调查方法所需的过渡到独立的地位，作为一个资助的研究人员，在实验肺病理学领域的兴趣和专业知识。成立了一个咨询委员会，以帮助我的学术生涯发展。