Okumiya Toshika, Kroos Marian A, Vliet Laura Van, Takeuchi Hiroaki, Van der Ploeg Ans T, Reuser Arnold J J
Department of Clinical Genetics, Erasmus MC, PO Box 2040, 3000 CA Rotterdam, The Netherlands.
Mol Genet Metab. 2007 Jan;90(1):49-57. doi: 10.1016/j.ymgme.2006.09.010. Epub 2006 Nov 13.
Glycogen storage disease type II (GSDII; Pompe disease or acid maltase deficiency) is an autosomal recessive disorder caused by lysosomal acid alpha-glucosidase (AalphaGlu) deficiency and manifests predominantly as skeletal muscle weakness. Defects in post-translational modification and transport of mutant AalphaGlu species are frequently encountered and may potentially be corrected with chaperone-mediated therapy. In the present study, we have tested this hypothesis by using deoxynojirimycin and derivatives as chemical chaperones to correct the AalphaGlu deficiency in cultured fibroblasts from patients with GSDII. Four mutant phenotypes were chosen: Y455F/Y455F, P545L/P545L, 525del/R600C and D645E/R854X. In case of Y455F/Y455F and P545L/P545L, N-(n-butyl)deoxynojirimycin (NB-DNJ) restored the transport, maturation and activity of AalphaGlu in a dose dependent manner, while it had no effect on the reference enzyme beta-hexosaminidase. NB-DNJ promoted export from the endoplasmic reticulum (ER) to the lysosomes and stabilized the activity of mutant AalphaGlu species, Y455F and P545L, inside the lysosomes. In long-term culture, the AalphaGlu activity in the fibroblasts from the patients with mutant phenotypes, Y455F/Y455F and P545L/P545L, increased up to 14.0- and 7.9-fold, respectively, in the presence of 10mumol/L NB-DNJ. However, the effect of NB-DNJ on Y455F/Y455F subsided quickly after removal of the compound. We conclude that NB-DNJ acts in low concentration as chemical chaperone for certain mutant forms of AalphaGlu that are trapped in the ER, poorly transported or labile in the lysosomal environment. Chemical chaperone therapy could create new perspectives for therapeutic intervention in GSDII.
II型糖原贮积病(GSDII;庞贝病或酸性麦芽糖酶缺乏症)是一种常染色体隐性疾病,由溶酶体酸性α-葡萄糖苷酶(AalphaGlu)缺乏引起,主要表现为骨骼肌无力。经常会遇到突变型AalphaGlu物种的翻译后修饰和转运缺陷,而伴侣介导的疗法可能会对其进行纠正。在本研究中,我们通过使用脱氧野尻霉素及其衍生物作为化学伴侣来纠正GSDII患者培养成纤维细胞中的AalphaGlu缺乏,从而验证了这一假设。选择了四种突变表型:Y455F/Y455F、P545L/P545L、525del/R600C和D645E/R854X。对于Y455F/Y455F和P545L/P545L,N-(正丁基)脱氧野尻霉素(NB-DNJ)以剂量依赖的方式恢复了AalphaGlu的转运、成熟和活性,而对参比酶β-己糖胺酶没有影响。NB-DNJ促进了从内质网(ER)到溶酶体的转运,并稳定了溶酶体内突变型AalphaGlu物种Y455F和P545L的活性。在长期培养中,在存在10μmol/L NB-DNJ的情况下,具有突变表型Y455F/Y455F和P545L/P545L的患者成纤维细胞中的AalphaGlu活性分别增加了14.0倍和7.9倍。然而,去除该化合物后,NB-DNJ对Y455F/Y455F的作用迅速减弱。我们得出结论,低浓度的NB-DNJ作为化学伴侣作用于某些被困在内质网中、转运不良或在溶酶体环境中不稳定的AalphaGlu突变形式。化学伴侣疗法可能为GSDII的治疗干预创造新的前景。