Department of Molecular Oral Medicine and Maxillofacial Surgery, Division of Applied Life Science, Graduate Institute of Biomedical and Health Science, Hiroshima University, 1-2-3, Kasumi, Minami-ku, Hiroshima-city, Hiroshima, 734-8553, Japan.
Mukasa Dental Clinic, Kanagawa, Japan.
Odontology. 2022 Jul;110(3):444-451. doi: 10.1007/s10266-021-00674-5. Epub 2021 Nov 15.
Cleidocranial dysplasia (CCD) is an autosomal dominant hereditary disease associated with the gene RUNX2. Disease-specific induced pluripotent stem cells (iPSCs) have emerged as a useful resource to further study human hereditary diseases such as CCD. In this study, we identified a novel CCD-specific RUNX2 mutation and established iPSCs with this mutation. Biopsies were obtained from familial CCD patients and mutation analyses were performed through Sanger sequencing and next generation sequencing. CCD-specific human iPSCs (CCD-hiPSCs) were established and maintained under completely defined serum, feeder, and integration-free condition using a non-integrating replication-defective Sendai virus vector. We identified the novel mutation RUNX2_c.371C>G and successfully established CCD-hiPSCs. The CCD-hiPSCs inherited the same mutation, possessed pluripotency, and showed the ability to differentiate the three germ layers. We concluded that RUNX2_c.371C>G was likely pathogenic because our results, derived from next generation sequencing, are supported by actual clinical evidence, familial tracing, and genetic data. Thus, we concluded that hiPSCs with a novel CCD-specific RUNX2 mutation are viable as a resource for future studies on CCD.
颅锁骨发育不全症(CCD)是一种常染色体显性遗传疾病,与 RUNX2 基因相关。疾病特异性诱导多能干细胞(iPSC)的出现为进一步研究 CCD 等人类遗传性疾病提供了有用的资源。在这项研究中,我们发现了一种新的 CCD 特异性 RUNX2 突变,并建立了带有该突变的 iPSC。通过桑格测序和下一代测序对家族性 CCD 患者进行活检和突变分析。使用非整合性复制缺陷型仙台病毒载体,在完全定义的血清、饲养层和无整合条件下建立和维持 CCD 特异性人 iPSC(CCD-hiPSC)。我们确定了新的 RUNX2_c.371C>G 突变,并成功建立了 CCD-hiPSC。CCD-hiPSC 继承了相同的突变,具有多能性,并表现出分化为三个胚层的能力。我们得出结论,RUNX2_c.371C>G 可能是致病性的,因为我们的结果来源于下一代测序,得到了实际临床证据、家族追踪和遗传数据的支持。因此,我们得出结论,具有新型 CCD 特异性 RUNX2 突变的 hiPSC 是未来研究 CCD 的可行资源。