真性红细胞增多症基因组学与疾病进展的关系

真性红细胞增多症基因组学与疾病进展的关系

赵迎旭¹,白洁²,张磊²,盛梦瑶³,石慧³,邢文³,杨逢春¹,艾丽梅⁴,周圆³

1. 1辽宁医学院附属一院血液科 2中国医学科学院北京协和医学院血液病医院（血液学研究所）
2. 中国医学科学院血液病医院
3. 中国医学科学院北京协和医学院血液病医院（血液学研究所）
4. 辽宁锦州辽宁医学院附属第一医院血液科

收稿日期:2014-02-11 修回日期:2014-04-18 出版日期:2014-06-15 发布日期:2014-06-15
通讯作者: 白洁

真性红细胞增多症基因组学与疾病进展的研究

Received:2014-02-11 Revised:2014-04-18 Published:2014-06-15 Online:2014-06-15
Contact: J Bai

赵迎旭1 ,白洁2 ,张磊2 ,盛梦瑶3 ,石慧3 ,邢文3 ,杨逢春1 ,艾丽梅4 ,周圆3 . 真性红细胞增多症基因组学与疾病进展的关系[J]. .

摘要/Abstract

摘要： 【摘要】目的研究135例真性红细胞增多症(PV)患者ASXL1、TET2、IDH1、IDH2、SETBP1和MPL515 、JAK2V617F、JAK2 exon12、基因突变情况，综合评判PV疾病进展与基因组学的特征。方法用Sanger方法对 ASXL1(Exon12)，TET2 (Exons 3-11)，IDH1(Exon4)，IDH2(Exon4)，SETBP1(Exon4)， JAK2 exon12和MPL515(Exon 10)等基因进行测序；用巢式等位基因特异性PCR方法检测JAK2V617F突变，对突变病例用Taqman-MGB探针行检测突变负荷。分析基因突变与疾病进展相关性。应用Kaplan-Meier单因素生存分析、Cox多因素分析和Logistic多因素分析等方法研究PV后继发骨髓纤维化（PPMF）的危险因素。结果PV患者ASXL1、TET2、IDH1、IDH2突变率分别为7.69%（8/104）、5.26% (1/19) 、0.08% (1/120)、和0.08% (1/121)。未发现PV患者中存在MPL、SETBP1突变。PV患者JAK2总突变率为82.22%（111/135）。其中JAK2exon12突变率为2.96%(4/135)。PPMF患者中，ASXL1突变率高达31.82%（7/22）。Spearman’s相关分析显示ASXL1与JAK2V617F负荷(V617F%)有相关性（r=0.298，P=0.002），ASXL1阳性患者，血红蛋白较低（p=0.049）。Kaplan-Meier生存分析显示，ASXL1 阳性(Log Rank =9.189, P=0.002) 、V617F% (Log Rank=9.339, P=0.002)、和WBC≥25×109/L (Log Rank =4.191, P=0.041)是PV患者无骨髓纤维化生存的危险因素。Cox多因素分析未能够得出PPMF的危险因素。但是Logistic多因素回归分析显示ASXL1突变 (β=3.47，Wald =3.932，OR=32.151，P=0.047，OR值95%CI为1.041-992.968)、V617F% (β=-1.582，Wald =5.157，OR=0.206，P=0.023，OR值95%CI为0.052-0.805)是PPMF的危险因素。结论我国PPMF患者具有更高的ASXL1突变率。ASXL1突变与高V617F%有相关性。ASXL1突变可能是PPMF的危险因素。ASXL-1突变可能通过某种机制，促进V617F%增高，参与到PPMF的发生和发展。PV患者TET2、IDH1、IDH2、SETBP1、MPL突变发生率较低。

关键词: 真性红细胞增多症, 基因组学, 疾病进展

Abstract: 【Abstract】Objective Mutation screening was performed for genes including ASXL1,TET2, IDH1, IDH2, SETBP1, MPL515, JAK2exon12 and JAK2V617, in 135 polycythemia vera (PV) patients. The correlation between the genomics characteristics and disease progression of PV were evaluated. Methods DNA sequencing of ASXL1(Exon12)，TET2 (Exons 3-11)，IDH1(Exon4)，IDH2(Exon4)，SEPBP1(Exon4)， JAK2exon 12 and MPL515(Exon 10) genes were carried out. The burden of mutant JAK2V617F allele (V617F%) was evaluated by real-time PCR in the meantime. The significative gene mutations and clinical outcomes or PPMF was analyzed. To study the risk factors of PPMF, Kaplan–Meier estimation, Cox and Logistic regression were applied. Results ASXL1、TET2、IDH1、IDH2 were mutated in 7.69%（8/104）、5.26% (1/19) 、0.08% (1/120)、and 0.08% (1/121) JAK2 was mutated in 82.22%(111/135) , among them, the mutation rate of JAK2exon12 was 2.96%(4/135) and there were no mutation of MPL515 and SETBP1 in these PV patients. The mutation of ASXL1 was 31.82%（7/22）in PPMF patients. The result of Spearman’s analysis showed that ASXL1 is correlated with V617F% (r=0.298，P=0.002). The hemoglobin was lower in patients with ASXL1 mutation. The result of Kaplan-Meier survival analysis showed ASXL1 mutation (Log Rank =9.189, P=0.002), V617F% higher than 50% (Log Rank=9.339,P=0.002) and leukocyte count >25×109/L(Log Rank=4.191,P=0.041) were the risk factors for survival of no PPMF. ASXL1 mutation(β=3.47，Wald =3.932，OR=32.151，P=0.047， 95%CI=1.041-992.968), V617F% higher than 50% (β=-1.582，Wald =5.157，OR=0.206，P=0.023， 95%CI=0.052-0.805) were the risk-factors of PPMF by multiple-factor analysis of Logistic was used. Conclusion The ASXL1 mutation was higher in Chinese. The ASXL1 mutation was correlation with V617F%. The ASXL1 mutation may is the risk-factor of PPMF. ASXL1 mutation may promote the V617F% by some mechanism. The mutation in the TET2, IDH1, IDH2, SEPTBP1 and MPL were rare.

Key words: Polycythemia vera, genomics, disease progression