Meinke Peter, Limmer Sarah, Hintze Stefan, Schoser Benedikt
Friedrich-Baur-Institute at the Department of Neurology, University Hospital, Ludwig-Maximilians-University Munich, Munich, Germany.
Ann Transl Med. 2019 Jul;7(13):277. doi: 10.21037/atm.2019.04.18.
Pompe disease is a neuromuscular disease caused by a deficiency of lysosomal acid alpha-glucosidase (GAA) which degrades glycogen, resulting in progressive accumulation of lysosomal glycogen, lysosomal swelling and rupture. In addition, mitochondrial abnormalities have been frequently observed in muscle biopsy specimens of Pompe patients. Enzyme replacement therapy (ERT) using alglucosidase alfa, a recombinant human GAA, is so far the only available therapy. We evaluated glycolysis and basal respiration in primary human myoblasts from patients with Pompe disease and in mouse myoblasts from GAA knockout mice before and after alglucosidase alfa treatment.
We tested patient-derived primary human myoblasts and immortalized GAA mouse myoblasts for GAA activity, glycolytic activity, and mitochondrial respiration before and after alglucosidase alfa treatment using enzyme activity assays and SeaHorse measurements.
A significant reduction in glycolysis (30%) and in mitochondrial respiration (50%) was observed in both, human and mouse GAA-deficient myoblasts. Treatment with alglucosidase alfa resulted in partial recovery of both metabolic pathways with some variability in human myoblasts.
Future assessments of treatment efficacy should include screening for the metabolic effects on both glycolysis and mitochondrial respiration in order to obtain a better read-out of the cellular energy metabolism.
庞贝病是一种神经肌肉疾病,由溶酶体酸性α-葡萄糖苷酶(GAA)缺乏引起,该酶可降解糖原,导致溶酶体糖原进行性积累、溶酶体肿胀和破裂。此外,在庞贝病患者的肌肉活检标本中经常观察到线粒体异常。使用重组人GAA阿糖苷酶α进行酶替代疗法(ERT)是目前唯一可用的治疗方法。我们评估了阿糖苷酶α治疗前后庞贝病患者原代人成肌细胞和GAA基因敲除小鼠成肌细胞的糖酵解和基础呼吸。
我们使用酶活性测定和海马测量法,在阿糖苷酶α治疗前后,对患者来源的原代人成肌细胞和永生化GAA小鼠成肌细胞进行GAA活性、糖酵解活性和线粒体呼吸测试。
在人和小鼠GAA缺陷的成肌细胞中均观察到糖酵解(30%)和线粒体呼吸(50%)显著降低。阿糖苷酶α治疗导致两种代谢途径部分恢复,人成肌细胞存在一定变异性。
未来治疗效果评估应包括筛查对糖酵解和线粒体呼吸的代谢影响,以便更好地了解细胞能量代谢情况。