Karageorgos L, Harmatz P, Simon J, Pollard A, Clements P R, Brooks D A, Hopwood John J
Lysosomal Diseases Research Unit, Department of Genetic Medicine, Women's and Children's Hospital, North Adelaide, Australia.
Hum Mutat. 2004 Mar;23(3):229-33. doi: 10.1002/humu.10313.
Mucopolysaccharidosis type VI (MPS VI), or Maroteaux-Lamy syndrome, is a lysosomal storage disorder caused by a deficiency of N-acetylgalactosamine-4-sulfatase (ARSB). Seven MPS VI patients were chosen for the initial clinical trial of enzyme replacement therapy. Direct sequencing of genomic DNA from these patients was used to identify ARSB mutations. Each individual exon of the ARSB gene was amplified by PCR and subsequently sequenced. Nine substitutions (c.289C>T [p.Q97X], c.629A>G [p.Y210C], c.707T>C [p.L236P], c.936G>T [p.W312C], c.944G>A [p.R315Q], c.962T>C [p.L321P], c.979C>T [p.R327X], c.1151G>A [p.S384N], and c.1450A>G [p.R484G]), two deletions (c.356_358delTAC [p.Y86del] and c.427delG), and one intronic mutation (c.1336+2T>G) were identified. A total of 7 out of the 12 mutations identified were novel (p.Y86del, p.Q97X, p.W312C, p.R327X, c.427delG, p.R484G, and c.1336+2T>G). Two of these novel mutations (p.Y86del and p.W312C) were expressed in Chinese hamster ovary cells and analyzed for residual ARSB activity and mutant ARSB protein. The two common polymorphisms c.1072G>A [p.V358M] and c.1126G>A [p.V376M] were identified among the patients, along with the silent mutation c.1191A>G. Cultured fibroblast ARSB mutant protein and residual activity were determined for each patient, and, together with genotype information, were used to predict the expected clinical severity of each MPS VI patient.
黏多糖贮积症 VI 型（MPS VI），又称马罗-拉米综合征，是一种由 N-乙酰半乳糖胺-4-硫酸酯酶（ARSB）缺乏引起的溶酶体贮积症。选择了 7 名 MPS VI 患者进行酶替代疗法的初步临床试验。对这些患者的基因组 DNA 进行直接测序以鉴定 ARSB 突变。通过聚合酶链反应（PCR）扩增 ARSB 基因的每个外显子，随后进行测序。鉴定出 9 个替换突变（c.289C>T [p.Q97X]、c.629A>G [p.Y210C]、c.707T>C [p.L236P]、c.936G>T [p.W312C]、c.944G>A [p.R315Q]、c.962T>C [p.L321P]、c.979C>T [p.R327X]、c.1151G>A [p.S384N] 和 c.1450A>G [p.R484G]）、2 个缺失突变（c.356_358delTAC [p.Y86del] 和 c.427delG）以及 1 个内含子突变（c.1336+2T>G）。在鉴定出的 12 个突变中，共有 7 个是新的（p.Y86del、p.Q97X、p.W312C、p.R327X、c.427delG、p.R484G 和 c.1336+2T>G）。其中两个新突变（p.Y86del 和 p.W312C）在中国仓鼠卵巢细胞中表达，并分析其 ARSB 残余活性和突变型 ARSB 蛋白。在患者中鉴定出两个常见的单核苷酸多态性 c.1072G>A [p.V358M] 和 c.1126G>A [p.V376M]，以及沉默突变 c.1191A>G。测定了每位患者培养的成纤维细胞 ARSB 突变蛋白和残余活性，并结合基因型信息，用于预测每位 MPS VI 患者预期的临床严重程度。
