Department of Pathology, University of Florida College of Medicine, Gainesville, Florida, USA.
Department of Medicine, and University of Florida College of Medicine, Gainesville, Florida, USA.
Cell Reprogram. 2020 Aug;22(4):179-188. doi: 10.1089/cell.2020.0012. Epub 2020 Jun 30.
Since it is extremely difficult to establish an animal model for human chromosomal abnormalities, induced pluripotent stem cells (iPSCs) provide a powerful alternative to study underlying mechanisms of these disorders and identify potential therapeutic interventions. In this study we established iPSCs from a young girl with a hemizygous deletion of Xq27.3-q28 who exhibited global developmental delay and intellectual disability from early in infancy. The deletion site on the X chromosome includes Fragile X Mental Retardation 1 (FMR1), the gene responsible for fragile X syndrome, which likely contributes to the patient's neurodevelopmental abnormalities. The FMR1 gene was expressed in approximately half of the iPSC clones we generated while it was absent in the other half due to the random inactivation of normal and abnormal X chromosomes. The normal or absent expression pattern of the FMR1 gene was not altered when the iPSCs were differentiated into neural progenitor cells (NPCs). Moreover, chromosome reactivating reagents such as 5-aza-2-deoxycytidine, trichostatin A, and UNC0638, were tested in an attempt to reactivate the suppressed FMR1 gene in affected iPSC-NPCs. The affected and control isogenic iPSCs developed in this study are ideal models with which to identify downstream consequences caused by the Xq27.3-q28 deletion and also to provide tools for high-throughput screening to identify compounds potentially improving the well-being of this patient population.
由于建立人类染色体异常的动物模型极其困难,诱导多能干细胞(iPSC)为研究这些疾病的潜在机制和鉴定潜在的治疗干预方法提供了有力的替代方法。在本研究中,我们从一位 Xq27.3-q28 半合子缺失的年轻女孩中建立了 iPSC,该女孩从婴儿早期开始表现出全面发育迟缓及智力障碍。X 染色体上的缺失部位包括脆性 X 智力低下 1 基因(FMR1),该基因是脆性 X 综合征的致病基因,可能导致患者的神经发育异常。我们生成的大约一半 iPSC 克隆中表达了 FMR1 基因,而另一半由于正常和异常 X 染色体的随机失活而不存在该基因。当 iPSC 分化为神经祖细胞(NPC)时,FMR1 基因的正常或缺失表达模式并未改变。此外,还测试了染色体再激活试剂,如 5-氮杂-2'-脱氧胞苷、曲古抑菌素 A 和 UNC0638,试图重新激活受影响的 iPSC-NPC 中被抑制的 FMR1 基因。本研究中建立的受影响和对照同基因 iPSC 是理想的模型,可用于鉴定 Xq27.3-q28 缺失引起的下游后果,并提供高通量筛选工具,以鉴定可能改善该患者群体福祉的化合物。