SNP单体型分析的不同策略应用于COL1A1成骨不全症家系PGT-M的临床研究  

Clinical application of different strategies of SNP haplotype analysis for PGT-M of families with osteogenesis imperfecta caused by COL1A1 mutations

作　　者：王洁 朱丽华[1] 陈林君[1] 林飞 刁振宇[1] 徐志鹏[1] 王珊珊[1] 张宁媛[1] WANG Jie;ZHU Li-hua;CHEN Lin-jun;LIN Fei;DIAO Zhen-yu;XU Zhi-peng;WANG Shan-shan;ZHANG Ning-yuan(Center of Reproductive Medicine,Nanjing Drum Tower Hospital,the Affiliated Hospital of Nanjing University Medical School,Nanjing 210000)

机构地区：[1]南京大学医学院附属鼓楼医院生殖医学中心,南京210000

出　　处：《生殖医学杂志》2021年第12期1569-1574,共6页Journal of Reproductive Medicine

基　　金：国家自然科学基金青年基金项目(81801533);中华医学会临床医学科研专项资金—生殖医学青年医师研究与发展项目(18010220751)。

摘　　要：目的探索单核苷酸多态性(SNP)单体型分析技术在预防成骨不全症(OI)发生和阻断OI的遗传缺陷的纵向传递的有效性。方法取三个家系的活检囊胚的滋养层细胞,采用多重置换扩增(MDA)方法进行全基因组扩增、一代测序联合基于二代测序(NGS)的SNP单体型分析技术,对三个携带COL1A1基因致病突变的OI家系进行胚胎植入前单基因病遗传学检测(PGT-M)。家系Ⅰ、家系Ⅱ和家系Ⅲ分别依据先证者、溯源的家族样本和新发突变患者单精子的遗传信息构建SNP单体型。依据检测结果筛选基因型正常的整倍体胚胎进行移植,于孕中期行羊膜腔穿刺,对胎儿基因型进行产前诊断,在新生儿出生后持续随访。结果家系Ⅰ活检4枚囊胚,一代测序检测和SNP单体型分析结果均显示有1枚囊胚基因型正常且染色体拷贝数正常,移植后,在妊娠37周活产一名体重为3500 g健康男婴。家系Ⅱ活检3枚囊胚,PGT-M结果显示有2枚囊胚基因型正常,但其中1枚为11号染色体三体,移植基因型正常的整倍体囊胚后,未妊娠。家系Ⅲ活检3枚囊胚,PGT-M结果显示有2枚囊胚基因型正常,移植其中1枚胚胎后,在妊娠40周活产一名体重为4250 g健康女婴。结论在PGT-M中,可采用不同的SNP单体型分析策略使更多的单基因病患者从中获益。使用基于NGS的SNP单体型分析结合致病基因的一代测序检测技术,在植入前的胚胎期即能有效阻断单基因疾病的垂直传播,帮助患者生育健康孩子。Objective:To investigate the effect of single nucleotide polymorphism(SNP)haplotype analysis in preventing osteogenesis imperfecta(OI)and blocking the vertical transmission of OI genetic defects in families with COL1A1 pathogenic gene mutations.Methods:The trophoblast cells of blastocysts from three families were biopsied.The whole genome of trophoblast cells from blastocysts was amplified by multiple displacement amplification(MDA).Preimplantation genetic testing for monogenic disease(PGT-M)was performed in three OI families with pathogenic mutations of COL1A1 gene by first-generation sequencing combined with SNP haplotype analysis based on next generation sequencing(NGS).SNP haplotypes were constructed for familyⅠ,familyⅡand familyⅢbased on the genetic information of proband,traceable family samples and monosperma of patients with new mutations,respectively.The euploid embryos with normal genotype were selected for transfer according to the test results.Amniocentesis was performed in the second trimester of pregnancy to make prenatal diagnosis of fetal genotypes,and the newborns were followed up continuously after birth.Results:Four blastocysts were biopsied in familyⅠ.The results of first-generation sequencing and SNP haplotype analysis showed that one blastocyst had normal genotype and normal chromosome copy number.After transplantation,a healthy boy weighing 3500 g was born at 37 weeks of gestation.Three blastocysts were biopsied in family 2.PGT-M results showed that two blastocysts had normal genotype,but one of them was trisomy 11.After transplantation of euploid blastocysts with normal genotype,no pregnancy occurred.Three blastocysts were biopsied in family 3.PGT-M results showed that two blastocysts had normal genotypes.After transplantation of one of the blastocysts,a healthy female infant weighing 4250 g was born at 40 weeks of gestation.Conclusions:In PGT-M,more patients with monogenic gene defects can benefit from different strategies of SNP haplotype analysis.Usage of NGS-based haplotyping c

关 键 词：胚胎植入前遗传学检测 成骨不全症 COL1A1基因 SNP单体型分析 

分 类 号：R715.5[医药卫生—妇产科学] R711.6[医药卫生—临床医学]