Reichle A, Rothe G, Krause S, Zaiss M, Ullrich H, Schmitz G, Andreesen R
Department of Hematology and Oncology, University of Regensburg, Germany.
Leukemia. 1999 Aug;13(8):1227-34. doi: 10.1038/sj.leu.2401481.
Dose escalation during consolidation therapy of de novo AML, including myeloablative chemotherapy supported with autologous peripheral blood stem cell transplantation (aPBSCT), continuously improved outcome. Therefore, quality control of transplants is getting increasing interest. We studied leukapheresis products (LPs), consecutively collected during postremission treatment of 20 patients with de novo AML for minimal residual disease (MRD) by 5-parametric flow cytometry and for myelodysplasia (MDS)-associated alterations by paired lineage-selected colony assays for colony-forming units-megakaryocytes (CFU-mega) and burst-granulocytes-monocytes colony-forming units (CFU) to evaluate the predictive value of these transplant-associated parameters on outcome. We defined the leukemia-associated immunophenotype at diagnosis and studied the impact of MRD detection in LPs collected after double induction with TAD (thioguanine, daunorubicin, cytarabine) and HAM (mitoxantrone, high-dose cytarabine, n=18 patients) and TAD consolidation treatment (n=20 patients) on outcome after aPBSCT. The level of MRD in the transplants correlated with the relapse-free survival (RFS) using a cut-off level of 1 x 10(-3) residual leukemic cells. The median RFS was 6 months for the group with > or = 1 x 10(-3) residual leukemic cells and has not been reached in the group with low MRD levels (< 1 x 10(-3)). By using the same cut-off level a weak correlation could also be demonstrated between MRD in the pregraft bone marrow and RFS (P = 0.04). Quantitatively abnormal megakaryocytic colony growth in the back-up LPs collected after double induction and in the transplant LPs was characterized by the ratio CFU-mega/CFU. In the group of relapsing patients the ratio CFU-mega/CFU was significantly lower than in the group of patients with CCR (P = 0.004), both in the back-ups and in the transplants. All patients with CFU-mega/CFU ratios < 0.12 relapsed, five of seven patients developed MDS before progressing to full leukemic relapse. Using the optimized cut-off level for the ratio CFU-mega/CFU (< vs > or = 0.12), seven of 10 relapsing patients (70%) could be identified to be at risk of relapse, whereas MRD in the transplants identified only 50% of the relapses and MRD in the pregraft bone marrow 25%. In conclusion, the study could identify two pretransplant risk factors predicting relapse in patients with AML receiving aPBSCT in first CR: MRD in transplants as well as MDS-like alterations within the transplants. These results may have multifold implications on the design of risk-adapted chemotherapy as well as on purging techniques and may contribute to a better understanding of leukemogenesis.
初治急性髓系白血病（AML）巩固治疗期间的剂量递增，包括采用自体外周血干细胞移植（aPBSCT）支持的清髓性化疗，持续改善了治疗结果。因此，移植的质量控制越来越受到关注。我们研究了20例初治AML患者缓解后治疗期间连续采集的白细胞单采产物（LPs），通过5参数流式细胞术检测微小残留病（MRD），并通过针对巨核细胞集落形成单位（CFU-mega）和爆式粒细胞-单核细胞集落形成单位（CFU）的配对谱系选择集落测定法检测与骨髓增生异常综合征（MDS）相关的改变，以评估这些与移植相关参数对治疗结果的预测价值。我们定义了诊断时的白血病相关免疫表型，并研究了在接受TAD（硫鸟嘌呤、柔红霉素、阿糖胞苷）和HAM（米托蒽醌、高剂量阿糖胞苷，n = 18例患者）双重诱导以及TAD巩固治疗（n = 20例患者）后采集的LPs中MRD检测对aPBSCT后治疗结果的影响。移植中MRD水平与无复发生存期（RFS）相关，残余白血病细胞的临界值为1×10⁻³。残余白血病细胞≥1×10⁻³的组中位RFS为6个月，而MRD水平低（<1×10⁻³）的组尚未达到中位RFS。使用相同的临界值，也可证明移植前骨髓中的MRD与RFS之间存在弱相关性（P = 0.04）。双重诱导后采集的备用LPs和移植LPs中，巨核细胞集落生长定量异常的特征是CFU-mega/CFU比值。在复发患者组中，CFU-mega/CFU比值在备用LPs和移植LPs中均显著低于持续完全缓解（CCR）患者组（P = 0.004）。所有CFU-mega/CFU比值<0.12的患者均复发，7例患者中有5例在进展为完全白血病复发前发生了MDS。使用CFU-mega/CFU比值的优化临界值（<与≥0.12），10例复发患者中有7例（70%）可被确定有复发风险，而移植中的MRD仅能识别50%的复发，移植前骨髓中的MRD能识别25%的复发。总之，该研究可识别出两个预测首次完全缓解（CR）期接受aPBSCT的AML患者复发的移植前风险因素：移植中的MRD以及移植内类似MDS的改变。这些结果可能对风险适应性化疗的设计、净化技术有多重影响，并可能有助于更好地理解白血病发生机制。
