原发性肥大性骨关节病1例患儿的遗传学分析  

Genetic analysis of a child with Primary hypertrophic osteoarthropathy

作　　者：王陈[1] 邱雪平[1] 程亚婷 李博宇 张元珍 马建鸿 郑芳[1,2] Wang Chen;Qiu Xueping;Cheng Yating;Li Boyu;Zhang Yuanzhen;Ma Jianhong;Zheng Fang(Center for Gene Diagnosis&Department of Laboratory Medicine,Zhongnan Hospital of Wuhan University,Wuhan,Hubei 430071,China;Clinical Research Center for Prenatal Diagnosis and Birth Health of Hubei Province,Wuhan,Hubei 430071,China;Reproductive Medicine Center,Zhongnan Hospital of Wuhan University,Wuhan,Hubei 430071,China)

机构地区：[1]武汉大学中南医院基因诊断中心&检验科,武汉430071 [2]湖北省产前诊断与优生临床医学研究中心,武汉430071 [3]武汉大学中南医院生殖医学中心,武汉430071

出　　处：《中华医学遗传学杂志》2024年第9期1100-1104,共5页Chinese Journal of Medical Genetics

摘　　要：目的探讨1例原发性肥大性骨关节(PHO)病患儿的遗传学病因。方法选取2021年7月27日于武汉大学中南医院就诊的1例患儿为研究对象。采集患儿及其父母的外周血样,提取基因组DNA,进行全外显子组测序,对疑似的剪接变异进行Sanger测序家系验证,并通过minigene实验进行体外功能验证;采用长片段PCR对疑似的外显子缺失变异进行验证。本研究通过武汉大学中南医院医学伦理委员会的审查(伦理号:[2021]伦审字62号)。结果WES结果显示患儿携带HPGD基因复合杂合变异,包括父源性杂合缺失变异(Exon 3 del)和母源性杂合剪接变异(c.421+1G>T)。长片段PCR验证了患儿及其父亲存在包含第3外显子在内的7565 bp片段的杂合缺失(c.218-1304_324+6156del),minigene实验结果显示母源性的剪接变异导致了第4外显子的跳跃剪接。结论HPGD基因c.218-1304_324+6156del杂合缺失变异和c.421+1G>T杂合剪接变异可能是该患儿的致病原因,上述结果丰富了HPGD基因的变异谱,为受累家庭的遗传咨询和产前诊断提供了依据。ObjectiveTo explore the genetic etiology of a child with primary hypertrophic osteoarthropathy(PHO).MethodsA child who was admitted to Zhongnan Hospital of Wuhan University on July 27,2021 was selected as the study subject.Genomic DNA was extracted from peripheral blood samples of the child and his parents and subjected to whole exome sequencing.Suspected splicing variant was verified by Sanger sequencing of family members.In vitro function was validated through a minigene assay,whilst the suspected exonic deletion was validated by long-fragment PCR.This study was approved by the Children′s Hospital Affiliated to Zhengzhou University(Ethics No.2023-K-011).ResultsWhole exome sequencing revealed that the child has harbored compound heterozygous variants of HPGD gene,including a heterozygous deletion(Exon 3 del)derived from his father and a splicing variant(c.421+1G>T)derived from his mother.Long-fragment PCR verified that the child and his father had both harbored a 7565 bp heterozygous deletion(c.218-1304_324+6156del),whilst the minigene assay proved that the splicing variant has resulted in skipping of exon 4.ConclusionThe heterozygous c.218-1304_324+6156del deletion and the c.421+1G>T splicing variant of the HPGD gene probably underlay the pathogenesis in this child.Above finding has enriched the mutational spectrum of the HPGD gene and provided a basis for genetic counseling and prenatal diagnosis for this family.

关 键 词：原发性肥大性骨关节病 全外显子组测序 HPGD基因 Minigene实验 

分 类 号：R726.8[医药卫生—儿科]