Fernandez-L Africa, Sanz-Rodriguez Francisco, Zarrabeitia Roberto, Pérez-Molino Alfonso, Hebbel Robert P, Nguyen Julia, Bernabéu Carmelo, Botella Luisa-Maria
Centro de Investigaciones Biologicas, CSIC, Ramiro de Maeztu, 9. Madrid, Spain.
Cardiovasc Res. 2005 Nov 1;68(2):235-48. doi: 10.1016/j.cardiores.2005.06.009. Epub 2005 Jul 5.
Hereditary haemorrhagic telangiectasia (HHT) is originated by mutations in endoglin (HHT1) and ALK1 (HHT2) genes. The purpose of this work was to isolate and characterize circulating endothelial cells from HHT patients.
Pure primary cultures of blood outgrowth endothelial cells (BOECs) were obtained from 50 ml of peripheral blood by selection on collagen plates with endothelial medium.
The amount of endoglin in HHT1-BOECs is half the controls, but HHT2-BOECs are also endoglin-deficient. Since the TGF-beta/ALK1 pathway activates the endoglin promoter activity, these results suggest the involvement of ALK1 in endoglin gene expression. Endothelial TGF-beta pathways, mediated by ALK1 and ALK5, are impaired in HHT cells. HHT-BOECs show disorganized and depolymerized actin fibers, as compared to the organized stress fibers of healthy-BOECs. Functionally, HHT-BOECs have impaired tube formation, in contrast with the cord-like structures derived from normal donors.
Decreased endoglin expression, impaired TGF-beta signalling, disorganized cytoskeleton, and failure to form cord-like structures are common characteristics of endothelial cells from HHT patients. These features may lead to fragility of small vessels and bleeding characteristic of the HHT vascular dysplasia and to a disrupted and abnormal angiogenesis, which may explain the clinical symptoms associated with this disease.
遗传性出血性毛细血管扩张症(HHT)由内皮糖蛋白(HHT1)和激活素受体样激酶1(ALK1,HHT2)基因的突变引起。本研究的目的是分离并鉴定HHT患者的循环内皮细胞。
通过在含内皮细胞培养基的胶原平板上筛选,从50毫升外周血中获得纯的血源性内皮细胞(BOECs)原代培养物。
HHT1-BOECs中内皮糖蛋白的含量是对照组的一半,但HHT2-BOECs也缺乏内皮糖蛋白。由于转化生长因子-β(TGF-β)/ALK1信号通路激活内皮糖蛋白启动子活性,这些结果提示ALK1参与内皮糖蛋白基因表达。由ALK1和ALK5介导的内皮细胞TGF-β信号通路在HHT细胞中受损。与健康BOECs中有序的应力纤维相比,HHT-BOECs显示出肌动蛋白纤维紊乱和解聚。在功能上,与来自正常供体的索状结构相比,HHT-BOECs形成管腔的能力受损。
内皮糖蛋白表达降低、TGF-β信号传导受损、细胞骨架紊乱以及无法形成索状结构是HHT患者内皮细胞的共同特征。这些特征可能导致小血管的脆弱性以及HHT血管发育异常的出血特征,并导致血管生成中断和异常,这可能解释了与该疾病相关的临床症状。