Alari Valentina, Russo Silvia, Rovina Davide, Garzo Maria, Crippa Milena, Calzari Luciano, Scalera Claudia, Concolino Daniela, Castiglioni Elisa, Giardino Daniela, Prosperi Ennio, Finelli Palma, Gervasini Cristina, Gowran Aoife, Larizza Lidia
Istituto Auxologico Italiano-IRCCS, Medical Cytogenetics & Human Molecular Genetics, Milan, Italy.
Centro Cardiologico Monzino-IRCCS, Unit of Vascular Biology and Regenerative Medicine, Milan, Italy.
Stem Cell Res. 2019 Oct;40:101553. doi: 10.1016/j.scr.2019.101553. Epub 2019 Aug 28.
Rubinstein-Taybi syndrome (RSTS) is a neurodevelopmental disorder characterized by growth retardation, skeletal anomalies and intellectual disability, caused by heterozygous mutations in either CREBBP (RSTS1) or EP300 (RSTS2) genes. We characterized 3 iPSC lines generated by Sendai from blood of RSTS1 patients with unique non sense c.4435G > T, p.(Gly1479*), c.3474G > A, p.(Trp1158*) and missense c.4627G > T, p.(Asp1543Tyr) CREBBP mutations. All lines displayed iPSC morphology, pluripotency markers, trilineage differentiation potential, stable karyotype and specific mutations. Western-blot using a CREB-Binding Protein N-terminus antibody demonstrated the same amount of full length protein as control in the missense mutation line and reduced amount in lines with stop mutations.
鲁宾斯坦-泰比综合征（RSTS）是一种神经发育障碍，其特征为生长发育迟缓、骨骼异常和智力残疾，由CREBBP（RSTS1）或EP300（RSTS2）基因的杂合突变引起。我们对由仙台病毒从RSTS1患者血液中生成的3个诱导多能干细胞（iPSC）系进行了特征分析，这些患者具有独特的无义突变c.4435G > T，p.（Gly1479*）、c.3474G > A，p.（Trp1158*）以及错义突变c.4627G > T，p.（Asp1543Tyr）的CREBBP突变。所有细胞系均表现出iPSC形态、多能性标志物、三系分化潜能、稳定的核型以及特定突变。使用CREB结合蛋白N端抗体进行的蛋白质免疫印迹分析表明，在错义突变细胞系中，全长蛋白的量与对照相同，而在有终止突变的细胞系中，全长蛋白的量减少。
