Laboratoire de Biochimie Médicale, Rouen University Hospital and EA 4309, University of Rouen, Rouen, France.
Autophagy. 2010 Jul;6(5):589-99. doi: 10.4161/auto.6.5.11943. Epub 2010 Jul 1.
Fabry disease is a lysosomal storage disorder (LSD) caused by a deficiency in α-galactosidase A. The disease is characterized by severe major organ involvement, but the pathologic mechanisms responsible have not been elucidated. Disruptions of autophagic processes have been reported for other LSDs, but have not yet been investigated in Fabry disease. Renal biopsies were obtained from 5 adult male Fabry disease patients before and after 3 years of enzyme replacement therapy (ERT) with agalsidase alfa. Vacuole accumulation was seen in renal biopsies from all patients compared with control biopsies. Decreases in the number of vacuoles were seen after 3 years of ERT primarily in renal endothelial cells and mesangial cells. Measurement of the levels of LC3, a specific autophagy marker, in cultured cells from Fabry patients revealed increased basal levels compared to cells from non-Fabry subjects and a larger increase in response to starvation than seen in non-Fabry cells. Starvation in the presence of protease inhibitors did not result in a significant increase in LC3 in Fabry cells, whereas a further increase in LC3 was observed in non-Fabry cells, an observation that is consistent with impaired autophagic flux in Fabry disease. Overexpression of LC3 mRNA in Fabry fibroblasts compared to control cells is consistent with an upregulation of autophagy. Furthermore, LC3 and p62/SQSTM1 (that binds to LC3) staining in renal tissues and in cultured fibroblasts from Fabry patients supports impairment of autophagic flux. These findings suggest that Fabry disease is linked to a deregulation of autophagy.
法布里病是一种溶酶体贮积症(LSD),由α-半乳糖苷酶 A 缺乏引起。该疾病的特征是严重的主要器官受累,但尚未阐明其病理机制。其他 LSD 的自噬过程已经被报道,但尚未在法布里病中进行研究。肾脏活检取自 5 名成年男性法布里病患者,在接受阿加糖酶α酶替代治疗(ERT)3 年后进行。与对照活检相比,所有患者的肾脏活检中均可见空泡积聚。在 ERT 治疗 3 年后,主要在肾内皮细胞和系膜细胞中观察到空泡数量减少。对法布里病患者培养细胞中特定自噬标志物 LC3 的测量显示,与非法布里病患者的细胞相比,基础水平升高,对饥饿的反应更大,而非法布里病患者的细胞则没有这种反应。在蛋白酶抑制剂存在的情况下,饥饿不会导致法布里细胞中 LC3 的显著增加,而在非法布里细胞中观察到 LC3 的进一步增加,这一观察结果与法布里病中自噬通量受损一致。与对照细胞相比,法布里成纤维细胞中 LC3 mRNA 的过度表达与自噬的上调一致。此外,法布里病患者的肾脏组织和培养成纤维细胞中 LC3 和 p62/SQSTM1(与 LC3 结合)的染色支持自噬通量受损。这些发现表明,法布里病与自噬的失调有关。