Orho M, Bosshard N U, Buist N R, Gitzelmann R, Aynsley-Green A, Blümel P, Gannon M C, Nuttall F Q, Groop L C
Department of Endocrinology, Wallenberg Laboratory, Malmö University Hospital, University of Lund, 20502 Malmö, Sweden.
J Clin Invest. 1998 Aug 1;102(3):507-15. doi: 10.1172/JCI2890.
Glycogen storage disease type 0 (GSD-0) is a rare form of fasting hypoglycemia presenting in infancy or early childhood and accompanied by high blood ketones and low alanine and lactate concentrations. Although feeding relieves symptoms, it often results in postprandial hyperglycemia and hyperlactatemia. The glycogen synthase (GS) activity has been low or immeasurable in liver biopsies, whereas the liver glycogen content has been only moderately decreased. To investigate whether mutations in the liver GS gene (GYS2) on chromosome 12p12.2 were involved in GSD-0, we determined the exon-intron structure of the GYS2 gene and examined nine affected children from five families for linkage of GSD-0 to the GYS2 gene. Mutation screening of the 16 GYS2 exons was done by single-strand conformational polymorphism (SSCP) and direct sequencing. Liver GS deficiency was diagnosed from liver biopsies (GS activity and glycogen content). GS activity in the liver of the affected children was extremely low or nil, resulting in subnormal glycogen content. After suggestive linkage to the GYS2 gene had been established (LOD score = 2.9; P < 0.01), mutation screening revealed several different mutations in these families, including a premature stop codon in exon 5 (Arg246X), a 5'-donor splice site mutation in intron 6 (G+1T--> CT), and missense mutations Asn39Ser, Ala339Pro, His446Asp, Pro479Gln, Ser483Pro, and Met491Arg. Seven of the affected children carried mutations on both alleles. The mutations could not be found in 200 healthy persons. Expression of the mutated enzymes in COS7 cells indicated severely impaired GS activity. In conclusion, the results demonstrate that GSD-0 is caused by different mutations in the GYS2 gene.
0型糖原贮积病(GSD-0)是一种罕见的空腹低血糖症,出现在婴儿期或幼儿期,伴有高血酮以及低丙氨酸和乳酸浓度。尽管进食可缓解症状,但往往会导致餐后高血糖和高乳酸血症。在肝脏活检中,糖原合酶(GS)活性一直很低或无法测量,而肝脏糖原含量仅中度降低。为了研究12p12.2染色体上肝脏GS基因(GYS2)的突变是否与GSD-0有关,我们确定了GYS2基因的外显子-内含子结构,并检查了来自五个家庭的九名患病儿童,以确定GSD-0与GYS2基因的连锁关系。通过单链构象多态性(SSCP)和直接测序对16个GYS2外显子进行突变筛查。通过肝脏活检(GS活性和糖原含量)诊断肝脏GS缺乏症。患病儿童肝脏中的GS活性极低或为零,导致糖原含量低于正常水平。在确定与GYS2基因存在提示性连锁关系后(对数优势分数=2.9;P<0.01),突变筛查发现这些家庭中有几种不同的突变,包括外显子5中的一个提前终止密码子(Arg246X)、内含子6中的一个5'供体剪接位点突变(G+1T→CT)以及错义突变Asn39Ser、Ala339Pro、His446Asp、Pro479Gln、Ser483Pro和Met491Arg。七名患病儿童的两个等位基因均携带突变。在200名健康人中未发现这些突变。突变酶在COS7细胞中的表达表明GS活性严重受损。总之,结果表明GSD-0是由GYS2基因中的不同突变引起的。