Center for Medical Genetics, University Hospital Ghent, De Pintelaan 185, Ghent, Belgium.
Eur J Hum Genet. 2010 Aug;18(8):895-901. doi: 10.1038/ejhg.2010.45. Epub 2010 Apr 14.
Fibulin-4 is a member of the fibulin family, a group of extracellular matrix proteins prominently expressed in medial layers of large veins and arteries. Involvement of the FBLN4 gene in cardiovascular pathology was shown in a murine model and in three patients affected with cutis laxa in association with systemic involvement. To elucidate the contribution of FBLN4 in human disease, we investigated two cohorts of patients. Direct sequencing of 17 patients with cutis laxa revealed no FBLN4 mutations. In a second group of 22 patients presenting with arterial tortuosity, stenosis and aneurysms, FBLN4 mutations were identified in three patients, two homozygous missense mutations (p.Glu126Lys and p.Ala397Thr) and compound heterozygosity for missense mutation p.Glu126Val and frameshift mutation c.577delC. Immunoblotting analysis showed a decreased amount of fibulin-4 protein in the fibroblast culture media of two patients, a finding sustained by diminished fibulin-4 in the extracellular matrix of the aortic wall on immunohistochemistry. pSmad2 and CTGF immunostaining of aortic and lung tissue revealed an increase in transforming growth factor (TGF)beta signaling. This was confirmed by pSmad2 immunoblotting of fibroblast cultures. In conclusion, patients with recessive FBLN4 mutations are predominantly characterized by aortic aneurysms, arterial tortuosity and stenosis. This confirms the important role of fibulin-4 in vascular elastic fiber assembly. Furthermore, we provide the first evidence for the involvement of altered TGFbeta signaling in the pathogenesis of FBLN4 mutations in humans.
纤维连接蛋白 4 是纤维连接蛋白家族的一员,该家族是一组细胞外基质蛋白,在大静脉和动脉的中层中大量表达。在小鼠模型和 3 名患有伴有全身受累的弹性皮肤松弛症的患者中,发现 FBLN4 基因参与心血管病理学。为了阐明 FBLN4 在人类疾病中的作用,我们研究了两个患者队列。对 17 名弹性皮肤松弛症患者进行直接测序,未发现 FBLN4 突变。在第二组 22 名表现为动脉扭曲、狭窄和动脉瘤的患者中,在 3 名患者中发现了 FBLN4 突变,两种纯合错义突变(p.Glu126Lys 和 p.Ala397Thr)和错义突变 p.Glu126Val 和移码突变 c.577delC 的复合杂合性。免疫印迹分析显示两名患者的成纤维细胞培养基中纤维连接蛋白-4 蛋白量减少,免疫组化显示主动脉壁细胞外基质中纤维连接蛋白-4 减少,证实了这一发现。对主动脉和肺组织的 pSmad2 和 CTGF 免疫染色显示转化生长因子 (TGF)β信号增加。这通过成纤维细胞培养物的 pSmad2 免疫印迹得到证实。总之,隐性 FBLN4 突变患者主要表现为主动脉瘤、动脉扭曲和狭窄。这证实了纤维连接蛋白-4 在血管弹性纤维组装中的重要作用。此外,我们提供了第一个证据,证明 TGFβ信号的改变参与了人类 FBLN4 突变的发病机制。