ANO5非移码插入突变型GDD家系临床分析及其诱导多能干细胞的构建

doi:10.19438/j.cjoms.2024.01.006

中国口腔颌面外科杂志 ›› 2024, Vol. 22 ›› Issue (1): 36-41.doi: 10.19438/j.cjoms.2024.01.006

ANO5非移码插入突变型GDD家系临床分析及其诱导多能干细胞的构建

王周阳¹，凌霄¹，施燕妮¹，王磊²，秦兴军¹

1.上海交通大学医学院附属第九人民医院口腔颌面-头颈肿瘤科，上海交通大学口腔医学院，国家口腔医学中心，国家口腔疾病临床医学研究中心，上海市口腔医学重点实验室，上海市口腔医学研究所，上海 200011;
2.柏林自由大学生物化学系，德国柏林 14195

收稿日期:2023-06-26 修回日期:2023-09-01 出版日期:2024-01-20 发布日期:2024-02-05
通讯作者: 秦兴军，E-mail: qinxj1989@sina.com
作者简介:王周阳（1998-），男，硕士，E-mail: wangzy.work@foxmail.com
基金资助:
国家自然科学基金（82071099）；上海交通大学医学院附属第九人民医院交叉基金（JYJC202007）上海交通大学医学院附属第九人民医院罕见病专项基金（JYHJB01）

Clinical analysis of ANO5 non-frameshift insertion mutation GDD family and generation of the induced pluripotent stem cells

WANG Zhou-yang¹, LING Xiao¹, SHI Yan-ni¹, WANG Lei², QIN Xing-jun¹

1. Department of Oromaxillofacial Head and Neck Oncology, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine; College of Stomatology, Shanghai Jiao Tong University; National Center for Stomatology; National Clinical Research Center for Oral Diseases; Shanghai Key Laboratory of Stomatology. Shanghai 200011, China;
2. Institut für Biochemie, Freie Universit?t Berlin. Berlin 14195, Germany

Received:2023-06-26 Revised:2023-09-01 Online:2024-01-20 Published:2024-02-05

摘要/Abstract

摘要： 目的: 建立和鉴定临床发现的一个由ANO5基因新的致病性突变引起的颌骨骨干发育不良（gnathodiaphyseal dysplasia，GDD）家系中患病者和非患病者来源的诱导多能干细胞（induced pluripotent stem cells，iPSCs）。方法: 纳入以颌骨畸形为主要表现的1个GDD家系5例患者，评估患者临床表现、骨影像学、骨密度、骨量等相关生化指标。收集家系中1例患病者与非患病者外周血样本，将表达Oct4、Sox2和Klf4 转录因子的仙台病毒转染患病者和非患病者外周血单核细胞，将其重编程为iPSCs，并通过碱性磷酸酶染色、细胞免疫荧光、qPCR、核型分析等检测其干细胞多能性。结果: GDD主要表现为双侧上下颌骨弥漫性多发肿胀，并伴有全身骨密度下降。通过检测，证明患病者及非患病者外周血来源的iPSCs具有多能性。结论: GDD患者外周血单核细胞在体外可被成功诱导为无外源性基因整合的具有多能分化特性的iPSCs，为GDD的发病机制研究提供了高效直接的细胞模型。

关键词: 颌骨骨干发育不良, ANO5基因, 表型, 诱导多能干细胞

Abstract: PURPOSE: To generate and identify gnathodiaphyseal dysplasia(GDD) patient and healthy donor derived induced pluripotent stem cells from a Chinese GDD family caused by a mutation in ANO5 gene. METHODS: The clinical manifestations, skeletal radiographic features, bone mineral density (BMD), and bone turn over biomarkers were investigated of 5 patients from a Chinese GDD family with facial deformities. Peripheral blood mononuclear cells(PBMCs) from one patient and one non-patient were collected and transfected with sendai virus carrying Oct4, Sox2 and Klf4 to be reprogrammed into iPSCs. The pluripotency was tested by alkaline phosphatase staining, immune-fluorescence, qPCR, and karyotyping analysis. RESULTS: GDD was mainly manifested as diffuse expansive swelling of maxilla and mandible with reduced bone mineral density throughout the body. iPSCs derived from PBMCs of the patient and the healthy donor maintained pluripotency. CONCLUSIONS: PBMCs of GDD patient are successfully reprogrammed into integration-free iPSCs with the pluripotency. These iPSCs provide a valuable cell model for mechanism exploration.

Key words: Gnathodiaphyseal dysplasia, ANO5 gene, Phenotype, Induced pluripotent stem cells

中图分类号:

R739.8

王周阳，凌霄，施燕妮，王磊，秦兴军. ANO5非移码插入突变型GDD家系临床分析及其诱导多能干细胞的构建[J]. 中国口腔颌面外科杂志, 2024, 22(1): 36-41.

WANG Zhou-yang， LING Xiao， SHI Yan-ni， WANG Lei， QIN Xing-jun. Clinical analysis of ANO5 non-frameshift insertion mutation GDD family and generation of the induced pluripotent stem cells[J]. China J Oral Maxillofac Surg, 2024, 22(1): 36-41.

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ANO5非移码插入突变型GDD家系临床分析及其诱导多能干细胞的构建

Clinical analysis of ANO5 non-frameshift insertion mutation GDD family and generation of the induced pluripotent stem cells

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