Droplet Digital PCR Improves IG-/TR-based MRD Risk Definition in Childhood B-cell Precursor Acute Lymphoblastic Leukemia.
Droplet Digital PCR Improves IG-/TR-based MRD Risk Definition in Childhood B-cell Precursor Acute Lymphoblastic Leukemia.
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Droplet Digital PCR改善了儿童B细胞前体急性淋巴细胞白血病中基于IG/TR的MRD风险定义
DOI：
10.1097/hs9.0000000000000543
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发表时间：
2021-03
期刊：
影响因子：
6.6
通讯作者：
Cazzaniga G
中科院分区：
文献类型：
--
作者：
Della Starza I;Nunes V;Lovisa F;Silvestri D;Cavalli M;Garofalo A;Campeggio M;De Novi LA;Soscia R;Oggioni C;Mussolin L;Biondi A;Guarini A;Valsecchi MG;Conter V;Biffi A;Basso G;Foà R;Cazzaniga G
微小残留病(MRD)是影响儿童急性淋巴细胞白血病(ALL)预后的最重要因素。实时定量聚合酶链式反应(RQ-PCR)代表了分子MRD评估和基于风险的一线治疗分层的金标准。在意大利儿科血液与肿瘤学协会(AIEOP)和柏林-法兰克福-慕尼黑(BFM)组AIEOP-BFM ALL2009和ALL2017的方案中，B系所有+33天RQ-PCR-MRD高且+78天阳性的患者被定义为慢早期反应(SERS)。根据AIEOP-BFM ALL2000研究的结果，当+78天的阳性MRD信号低于RQ-PCR定量的下限(“阳性不可量化，”POS-NQ)时，这些患者也被视为高危患者。为了评估Droplet数字聚合酶链式反应(DdPCR)是否可以提高患者的风险定义，我们分析了AIEOP-BFM ALL2000试验中209例经RQ-PCR归类为POS-NQ和/或阴性(NEG)的儿童B系患者的MRD。对45例SER患者+78d采集的RQ-PCRMRD+33d和≥-NQ分别为5.0×10-4和+78d的样本进行DDPCRMRD分析。分析确定了45例可量化阳性病例中的13例。大多数复发发生在该患者的亚组中，而ddPCRNEG或ddPCRPOSNQ患者的预后明显好于后者(P<；0.001)。总体而言，在112例MR病例和52例标准风险患者中，除了少数病例没有结果差异外，ddPCR分析证实了MRD阴性和POS-NQ。这些数据表明，ddPCR在测量MRD方面比RQ-PCR更准确，特别是在后期的随访时间点，因此可以在所有方案中改善患者的分层。
Minimal residual disease (MRD) is the most powerful prognostic factor in pediatric acute lymphoblastic leukemia (ALL). Real-time quantitative polymerase chain reaction (RQ-PCR) represents the gold standard for molecular MRD assessment and risk-based stratification of front-line treatment. In the protocols of the Italian Association of Pediatric Hematology and Oncology (AIEOP) and the Berlin-Frankfurth-Munschen (BFM) group AIEOP-BFM ALL2009 and ALL2017, B-lineage ALL patients with high RQ-PCR-MRD at day+33 and positive at day+78 are defined slow early responders (SERs). Based on results of the AIEOP-BFM ALL2000 study, these patients are treated as high-risk also when positive MRD signal at day +78 is below the lower limit of quantification of RQ-PCR (“positive not-quantifiable,” POS-NQ). To assess whether droplet digital polymerase chain reaction (ddPCR) could improve patients’ risk definition, we analyzed MRD in 209 pediatric B-lineage ALL cases classified by RQ-PCR as POS-NQ and/or negative (NEG) at days +33 and/or +78 in the AIEOP-BFM ALL2000 trial. ddPCR MRD analysis was performed on 45 samples collected at day +78 from SER patients, who had RQ-PCR MRD ≥ 5.0 × 10–4 at day+33 and POS-NQ at day+78 and were treated as medium risk (MR). The analysis identified 13 of 45 positive quantifiable cases. Most relapses occurred in this patients’ subgroup, while ddPCR NEG or ddPCR-POS-NQ patients had a significantly better outcome (P < 0.001). Overall, in 112 MR cases and 52 standard-risk patients, MRD negativity and POS-NQ were confirmed by the ddPCR analysis except for a minority of cases, for whom no differences in outcome were registered. These data indicate that ddPCR is more accurate than RQ-PCR in the measurement of MRD, particularly in late follow-up time points, and may thus allow improving patients’ stratification in ALL protocols.