PK-LR基因新突变丙酮酸激酶缺乏症患儿经异基因造血干细胞移植治愈

Pyruvate Kinase Deficiency Children with New Types of PK-LR Gene Mutation Were Cured by Allogeneic Hematopoietic Stem Cell Transplantation

目的 通过对2例丙酮酸激酶缺乏症（PKD）患儿临床症状及PK-LR基因新突变类型的报道,探讨PKD的PK-LR基因诊断方法及行异基因造血干细胞移植（allo-HSCT）的疗效.方法 选择2009年2月及2013年8月,上海交通大学医学院附属上海儿童医学中心收治的2例PKD患儿作为研究对象.本研究遵循的程序符合上海交通大学医学院附属上海儿童医学中心人体试验委员会所制定的伦理学标准,得到该委员会批准,征得受试对象监护人的知情同意,并与之签署临床研究知情同意书.通过上海儿童医学中心检验科采集2例患儿的血液标本,采用Sanger基因测序及全外显子捕获测序对2例患儿PK-LR基因进行检测分析.采用allo-HSCT对2例患儿进行治疗,并分别对2例患儿进行随访.回顾性分析该2例患儿的临床症状、诊断方法及治疗方案.结果 ①2例PKD患儿丙酮酸激酶（PK）活性均降低,分别为8.00 IU/gHb与7.61 IU/gHb.②2例PKD患儿PK-LR基因检测分析共发现4种PK LR基因错义突变.其中,PK-LR基因c.941T＞C（p.Ile314Thr）突变已有文献报道,c.119G＞A（p.Arg40Gln）,c.1015G＞A（p.Asp339Asn）及c.848T＞C（p.Va1283Ala）突变为PK-LR基因新的突变类型.4种突变的SIFT功能预测结果分别为0.20,0.37,0及0.23.③2例PKD患儿均接受allo-HSCT.患儿1于移植后第14天粒系及红系造血功能获得恢复,移植后第19天血小板（PLT）计数＞5x10^1 0/L,移植后第22天造血干细胞（HSC） 100％嵌合,患儿血型亦转换为供者血型.患儿2于移植后第12天粒系及红系造血功能获得恢复,移植后第19天PLT计数＞3x10^10/L,移植后第18天HSC 100％嵌合,患儿血型亦转换为供者血型.④对2例PKD患儿分别随访5年和1年,患儿生存状况良好,造血系统及其他系统指标均为正常,获得成功治愈.结论 全外显子捕获基因测序方法可用于临床PKD基因诊断及PK-LR基因新型突变类型的发现;PK-LR基因新突变类型c.G119G＞A（p.Arg40Gln）,c.1015G＞A（p.Asp339Asn）及c.848T＞C（p.Va1283Ala）亦可导致PKD的发生;allo-HSCT治疗本研究中2例PKD患儿是有效、可行的.

Objective To investigate PK-LR genetic diagnosis and hematopoietic stem cell transplantation （allo-HSCT） of pyruvate kinase deficiency （PKD） through clinical symptoms of two children with PKD and new types of PK-LR gene mutation report.Methods In February 2009 and August 2013,two children with PKD in Shanghai Children＇s Medical Center,Affiliated to Shanghai Jiao Tong University School of Medicine were collected into this study.The study protocol was approved by the Ethical Review Board of Investigation in Human at Shanghai Children＇s Medical Center Affiliated to Shanghai Jiao Tong University School of Medicine.Informed consent was obtained from all participants＇ parents.The blood specimens from the two PKD children were collected.PK-LR gene analysis of two children were detected by Sanger sequencing and whole exon capture sepuencing.Two PKD children were treated by allo-HSCT,and their clinical symptoms,diagnosis and treatment were retrospectively analyzed.Results ① Pyruvate kinase （PK） activation of two PKD children were reduced to 8.00 IU/gHb and 7.61 IU/gHb respectively.② Four types of PK-LR gene missense mutation were found in gene detection of two children.PK-LR gene c.T941T〉C（p.Ile314Thr） mutation was reported formerly,and c.119G〉A （p.Arg40Gln）,c.1015G〉A（p.Asp339Asn） and c.848T〉 C （p.Va1283Ala） mutations were new.SIFT function prediction results of the 4 mutations were 0.20,0.37,0 and 0.23 respectively.③ Two PKD children were treated by alloHSCT.After transplantation,myeloid and erythroid hematopoiesis of children 1 were recovered on the 14th day.Platelet （PLT） count was 〉5＆#215; 1010/L on the 19th day.Hematopoietic stem cell （HSC） was complete chimerism on the 22th day,and the blood type of children 1 was converted to donor＆#39;s.After transplantation,myeloid and erythroid hematopoiesis of children 2 were recovered on the 12th day.PLT count was 〉3＆#215; 1010/L on the 19th day.HSC was complete chimerism on the 18th day,and the blood type of children 2 was converted to donor＆#39;s.Conclusions The whole exon capture sequencing can be used in the clinical diagnosis of PKD and searching for new types of PK-LR gene mutation.PK-LR gene c.119 G〉A （p.Arg40Gln）,c.1015G〉A（p.Asp339Asn） and c.848T〉C（p.Va1283Ala） missense mutations could also lead to PKD.allo-HSCT could be an effective treatment for PKD.