GENETIC HETEROGENEITY IN ENDOTHELIAL GENE EXPRESSION

这个项目解决了内皮细胞基因表达的遗传异质性。我们的主要假设是，血管内皮细胞基因表达的遗传差异将显著影响人类血管疾病的临床表型。两种相对较新的技术的独特结合使我们能够直接在人类身上实际测试这一概念：报告内皮细胞可以从外周血样本中以血液生长内皮细胞(BOEC)的形式获得，内皮细胞基因的表达可以通过微阵列图谱进行监测。这一假说的初步检验将集中在镰刀状贫血上，这是一种“单基因”疾病，在这种疾病中，两种非常不同的临床表型可以可靠地 已确定：卒中高危人群与非高危人群；高危人群将被细分为有Moya Moya者和无Moya Moya者。我们假设有中风风险的HbSS青少年(n=25)将表现出不同于无中风风险的镰刀受试者(n=25)的内皮细胞多态；我们还假设有Moya Moya的高危受试者(n=25)将表现出不同于无Moya Moya的高危受试者的多态。25名非镰刀种族和年龄匹配的青少年作为对照。在目标1中，BOEC基因表达的微阵列图谱将被用来鉴定构成内皮细胞的基因表达。在目标2中，我们将同时进行细胞生物学实验，以检查几种类型的内皮细胞反应性是否被认为是高概率的 与中风表型有关。在目标#3中，我们将验证候选基因或反应通路在目标#1和#2中的重要性。在目标#4中，我们将使用另一项正在进行的SNP研究的数据，该研究旨在寻找SNP与镰刀状中风的关联，为微阵列分析提供信息，从而将两种方法的力量结合在一起。

This project addresses genetic heterogeneity in endothelial gene expression. Our overarching hypothesis is that the clinical phenotype of human vascular diseases will be influenced significantly by inherited differences in gene expression by endothelial cells. The unique combination of two relatively new technologies allows us to actually test this concept, directly and in humans: reporter endothelial cells can be obtained from peripheral blood sampling in the form of blood outgrowth endothelial cells (BOEC), and endothelial gene expression can be monitored by microarray profiling. This initial test of the hypothesis will focus on sickle anemia, a "single gene" disorder in which two very distinct clinical phenotypes can be reliably identified: at-risk for stroke versus not-at-risk for stroke; the at risk-group will be subdivided into those with versus those without moya moya. W e hypothesize that adolescents with HbSS who are at-risk for stroke (n=25) will exhibit different endothelial polymorphisms than the sickle subjects who are not-at-risk for stroke (n=25 each); we also hypothesize that at-risk subjects with moya moya (n=25) will exhibit different polymorphisms from those without moya moya (n=25). 25 non-sickle race and age matched adolescents will serve as controls. In Aim #1, microarray profiling of BOEC gene expression will be used to identify constitutive endothelial gene expression. In Aim #2, we will in parallel perform cell biologic experiments to examine for exaggerated endothelial responsiveness of several types felt to be high-probability for involvement in the stroke phenotype. In Aim #3 we will verify significance of candidate gene or response pathways implicated in Aims #1 and #2. In Aim #4, we will use data from a separate ongoing SNP study that looks for SNP associations with sickle stroke, to inform microarray analysis and thereby combine power of the two approaches.