INSERM U920, Laboratoire des Mécanismes Moléculaires de l'Angiogenèse, Université Bordeaux 1, Talence, France.
PLoS One. 2009 Nov 17;4(11):e7856. doi: 10.1371/journal.pone.0007856.
Formation of blood vessels requires the concerted regulation of an unknown number of genes in a spatial-, time- and dosage-dependent manner. Determining genes, which drive vascular maturation is crucial for the identification of new therapeutic targets against pathological angiogenesis.
METHODOLOGY/PRINCIPAL FINDINGS: [corrected] We accessed global gene regulation throughout maturation of the chick chorio-allantoic membrane (CAM), a highly vascularized tissue, using pan genomic microarrays. Seven percent of analyzed genes showed a significant change in expression (>2-fold, FDR<5%) with a peak occurring from E7 to E10, when key morphogenetic and angiogenic genes such as BMP4, SMO, HOXA3, EPAS1 and FGFR2 were upregulated, reflecting the state of an activated endothelium. At later stages, a general decrease in gene expression occurs, including genes encoding mitotic factors or angiogenic mediators such as CYR61, EPAS1, MDK and MYC. We identified putative human orthologs for 77% of significantly regulated genes and determined endothelial cell enrichment for 20% of the orthologs in silico. Vascular expression of several genes including ENC1, FSTL1, JAM2, LDB2, LIMS1, PARVB, PDE3A, PRCP, PTRF and ST6GAL1 was demonstrated by in situ hybridization. Up to 9% of the CAM genes were also overexpressed in human organs with related functions, such as placenta and lung or the thyroid. 21-66% of CAM genes enriched in endothelial cells were deregulated in several human cancer types (P<.0001). Interfering with PARVB (encoding parvin, beta) function profoundly changed human endothelial cell shape, motility and tubulogenesis, suggesting an important role of this gene in the angiogenic process.
CONCLUSIONS/SIGNIFICANCE: Our study underlines the complexity of gene regulation in a highly vascularized organ during development. We identified a restricted number of novel genes enriched in the endothelium of different species and tissues, which may play crucial roles in normal and pathological angiogenesis.
血管的形成需要在空间、时间和剂量依赖的方式下协同调节未知数量的基因。确定驱动血管成熟的基因对于鉴定针对病理性血管生成的新治疗靶点至关重要。
方法/主要发现：我们使用全基因组微阵列研究了鸡绒毛尿囊膜（CAM）成熟过程中的整体基因调控，CAM 是一种高度血管化的组织。分析的 7%的基因表达发生显著变化（>2 倍， FDR<5%），峰值出现在 E7 到 E10 期间，此时关键的形态发生和血管生成基因如 BMP4、SMO、HOXA3、EPAS1 和 FGFR2 上调，反映了激活的内皮细胞状态。在后期，基因表达普遍下降，包括编码有丝分裂因子或血管生成介质的基因，如 CYR61、EPAS1、MDK 和 MYC。我们确定了 77%的显著调控基因的人类同源基因，并通过计算机预测了 20%的同源基因在血管内皮细胞中的富集情况。通过原位杂交证实了包括 ENC1、FSTL1、JAM2、LDB2、LIMS1、PARVB、PDE3A、PRCP、PTRF 和 ST6GAL1 在内的几个基因在血管中的表达。CAM 中的多达 9%的基因在具有相关功能的人类器官中也过表达，如胎盘和肺或甲状腺。21-66%在血管内皮细胞中富集的 CAM 基因在几种人类癌症类型中失调（P<.0001）。干扰 PARVB（编码 parvin，beta）的功能会极大地改变人内皮细胞的形状、运动性和管状形成，表明该基因在血管生成过程中起着重要作用。
结论/意义：我们的研究强调了发育过程中高度血管化器官中基因调控的复杂性。我们鉴定了在不同物种和组织的内皮细胞中富集的少数新基因，这些基因可能在正常和病理性血管生成中发挥关键作用。
