基于二代测序技术分析TKI耐药慢性粒细胞白血病患者的基因突变特征

Analysis of mutation characteristics in TKI-resistant chronic myeloid leukemia patients based on next-generation sequencing technology

目的探讨酪氨酸激酶抑制剂(TKI)耐药的慢性粒细胞白血病(CML)患者基因突变特征和规律,及其与TKI耐药CML的关系。方法回顾性病例系列研究。回顾性分析2018年8月至2022年11月在南方医科大学南方医院和汕头大学医学院附属粤北人民医院就诊的TKI耐药CML患者的临床资料和二代测序结果,分析总体及不同疾病阶段患者基因突变情况。结果共收集60例TKI耐药的CML患者,患者年龄[M(Q1,Q3)]41.5岁(32岁,53岁);男性38例(63.33%),女性22例(36.67%);慢性期43例,进展期17例(加速期3例,急变期14例)。共30例(50.00%)检测出非ABL1基因突变,包括45次15个非ABL1基因;60例患者中非ABL1突变基因数为1个(0个,2个)。60例患者中,ASXL1突变21例(35.00%),DNMT3A和RUNX1突变各5例(8.33%),SETBP1突变3例(5.00%);检测出1个和≥2个非ABL1基因突变患者分别占33.33%(20/60)和16.67%(10/60)。进展期和慢性期患者非ABL1基因突变总检出率分别为52.94%(9/17)、48.84%(21/43),≥2个非ABL1基因突变的检出率分别为23.53%(4/17)、13.95%(6/43),但差异均无统计学意义(χ2=0.08,P=0.774;χ2=0.80,P=0.370)。60例患者中17例(28.33%)ABL1激酶区突变,其中14例(82.35%)非ABL1基因突变;这17例中,6例进展期患者均非ABL1基因突变,11例慢性期患者中8例非ABL1基因突变,但差异无统计学意义(P=0.452)。结论TKI耐药的CML患者非ABL1基因突变率较高,并且进展期患者的突变率有高于慢性期患者的趋势,可能与耐药CML患者的BCR-ABL1融合基因异常激活ABL1激酶,导致基因组和表观基因组进一步突变,驱动疾病由慢性期向加速期或急变期进展有关。

Objective To explore the characteristics and patterns of gene mutations in tyrosine kinase inhibitor(TKI)-resistant chronic myeloid leukemia(CML)patients and their relationship with TKI-resistant CML.Methods A retrospective case series study was performed.Clinical data and next-generation sequencing results from TKI-resistant CML patients in Nanfang Hospital of Southern Medical University and Yuebei People's Hospital of Shantou University Medical College from August 2018 to November 2022 were retrospectively analyzed,and the gene mutations of the patients in general and at different disease stages were analyzed.ResultsSixty patients were enrolled,with the age[M(Q 1,Q 3)]of 41.5 years old(32 years old,53 years old);38 cases(63.33%)were male and 22 cases(36.67%)were female;43 cases were in the chronic stage,and 17 cases were in the progression stage(3 cases were in the accelerated stage and 14 cases were in the blast stage).non-ABL1 mutations were detected in 30 patients(50.00%)including 45 times of 15 non-ABL1 genes.The number of non-ABL1 mutation gene was 1(0,2)in 60 patients.Of the 60 patients,21(35.00%)had ASXL1 mutations,5(8.33%)had DNMT3A mutations,5(8.33%)had RUNX1 mutations,and 3(5.00%)had SETBP1 mutations;the proportions of patients with 1 and≥2 non-ABL1 mutations were 33.33%(20/60)and 16.67%(10/60),respectively.The total detection rates of non-ABL1 mutations were 52.94%(9/17)and 48.84%(21/43),and the detection rates of≥2 non-ABL1 mutations were 23.53%(4/17)and 13.95%(6/43)in patients with progression and patients with chronic disease,and the differences were not statistically significant(χ2=0.08,P=0.774;χ2=0.80,P=0.370).Seventeen of 60 patients(28.33%)had mutations in the ABL1 kinase region,of which 14(82.35%)had non-ABL1 mutations;of these 17 cases,6 patients with progressive disease all had non-ABL1 mutations,in 11 patients with chronic disease,8 patients had non-ABL1 mutations,and the difference was not statistically significant(P=0.452).Conclusions Patients with TKI-resistant CML have high frequencies of non-ABL1 mutations,and there is a trend for higher mutation rates in patients with progressive disease than in patients with chronic disease,and these may be related to the abnormal activation of ABL1 kinase by BCR-ABL1 fusion gene in patients with drug-resistant CML,which leads to the genome-level and epigenome-level mutations,and driving disease progression from chronic phase to accelerated or blast phase.