Donor-Derived CD19-Targeted T Cell Infusion Eliminates B Cell Acute Lymphoblastic Leukemia Minimal Residual Disease with No Response to Donor Lymphocytes after Allogeneic Hematopoietic Stem Cell Transplantation

Leukemia relapse is still the leading cause of treatment failure after allogeneic hematopoietic stem cell transplantation (allo-HSCT) for B cell acute lymphoblastic leukemia (B-ALL). Relapsed patients with BALL after allo-HSCT have a very short median survival. Minimal residual disease (MRD) is predictive of forthcoming hematological relapse after hematopoietic stem cell transplantation (HSCT);furthermore, eliminating MRD effectively prevents relapse. Donor lymphoblastic infusion (DLI) is the main established approach to treat B-ALL with MRD after allo-HSCT. However, about one-third of patients with MRD are non-responsive to DLI and their prognosis worsens. Although donor-derived cluster of differentiation (CD)19-directed chimeric antigen receptor-modified (CAR) T cells (CART19s) can potentially cure leukemia, the efficiency and safety of infusions with these cells have not yet been investigated in patients with MRD after HSCT. Between September 2014 and February 2018, six patients each received one or more infusions of CART19s from HSCT donors. Five (83.33%) achieved MRD-negative remission, and one case was not responsive to the administration of CAR T cells. Three of the six patients are currently alive without leukemia. No patient developed acute graft-versus-host disease (aGVHD), and no patient died of cytokine release syndrome. Donor-derived CAR T cell infusions seem to be an effective and safe intervention for patients with MRD in B-ALL after allo-HSCT and for those who were not responsive to DLI.

Study of Minimal Residual Disease in Adults with B-Lineage Acute Lymphoblastic Leukemia by Flowcytometry

Background: After achieving morphological remission, existence of few number of leukemic cells in the patient’s blood represents the minimal residual disease (MRD) and its monitoring helps in evaluating early treatment response and future relapse. Patients and methods: Eighty seven newly diagnosed (B-ALL) cases were enrolled in the present study in the time period from October 2013 to October 2016. A panel of 4 monoclonal antibodies (CD10FITC, CD19PE, CD34PercP and CD45APC) were defined at diagnosis and after morphological remission for tracing of minimal residual disease (MRD). Results: Eighty seven newly diagnosed B-ALL cases were included in the present study of which 73 (84%) showed positive expression to CD45 in combination with (CD10, CD19 and CD34) at diagnosis, which allow us to use this combination for further assessment of MRD after morphological remission. In our study 65% of patients had negative MRD ( Conclusion: MRD detection by flow cytometry using the combination of CD45 with CD10, CD19 & CD34 is an easy and reliable method. Patients with positive MRD are at higher risk of relapse and have inferior overall survival rates compared to those with MRD-ve. Future studies focusing on treatment intensification for the group of patients with +ve MRD aiming to improve the treatment outcome are warranted.

Carrimycin in the treatment of acute promyelocytic leukemia combined with pulmonary tuberculosis: A case report

BACKGROUND Pulmonary tuberculosis(PTB)is prevalent in immunocompromised populations,including patients with hematologic malignancies,human immunodeficiency virus infections,and chronic diseases.Effective treatment for acute promyelocytic leukemia(APL)combined with PTB is lacking.These patients show an extremely poor prognosis.Therefore,studies should establish efficient treatment options to improve patient survival and prognosis.CASE SUMMARY A 60-year-old male with pain in the right side of his chest and a fever for 4 d visited the outpatient department of our hospital.Peripheral blood smear revealed 54%blasts.Following bone marrow examinations,variant APL with TNRC18-RARA fusion gene was diagnosed.Chest computed tomography scan showed bilateral pneumonitis with bilateral pleural effusions,partial atelectasis in the lower lobes of both lungs,and the bronchoalveolar lavage fluid gene X-Pert test was positive,indicative of PTB.Carrimycin,ethambutol(EMB),and isoniazid(INH)were administered since he could not receive chemotherapy as the WBC count decreased continuously.After one week of treatment with carrimycin,the patient recovered from fever and received chemotherapy.Chemotherapy was very effective and his white blood cells counts got back to normal.After being given five months with rifampin,EMB and INH and chemotherapy,the patient showed complete remission from pneumonia and APL.CONCLUSION We report a case of PTB treated successfully with carrimycin with APL that requires chemotherapy.

A comparison of flow cytometry detection of minimal residual disease and chimerism kinetics in chronic lymphocytic leukemia patients after allogeneic hematopoietic stem cell transplantation

Determination of minimal residual disease (MRD) remains crucial for the follow-up after therapy in chronic lymphocytic leukemia (CLL) patients. Chimerism was assessed by short tandem repeat (STR)-PCR and single nucleotide polymorphisms (SNP)-PCR, and MRD by a multicolor flow cytometric approach in 12 consecutive patients with CLL after they received allogeneic stem cell transplantation (SCT). Overall, 11 patients achieved MRD flow negativity [10 had full donor chimerism (FDC) and one had mixed chimerism (MC)]. Only one patient remained with MRD flow positivity and displayed MC. Fifty-six samples were concomitantly studied by both chimerism and MRD flow. A significant correlation was observed between MRD flow data and chimerism in both PB and BM by using a mixed effect linear regression (p < 0.001). Flow cytometry approach of MRD can be easily combined with chimerism during the follow-up post-allogeneic SCT. Both techniques appeared complementary for guiding post-transplant immunomodulation.

Myeloid sarcoma of the colon as initial presentation in acute promyelocytic leukemia:A case report and review of the literature

BACKGROUND Myeloid sarcoma(MS)rarely occurs in acute promyelocytic leukemia(APL)at onset,but it can develop in relapse cases,especially after APL treated with alltrans retinoic acid(ATRA).Therefore little is known about the clinical features and suitable treatment for APL related MS due to the rarity of the disease,although this may be different from the treatment and prognosis of MS in the relapse stage.To our best knowledge,this is the second case report of APL initial presentation as colon MS.CASE SUMMARY A 77-year-old woman complained of intermittent right lower abdominal pain,black stool,and difficult defecation for 2 mo.Physical examination showed diffuse tenderness during deep palpation and an anemic appearance.Laboratory findings showed positivity for fecal occult blood testing;white blood cell count:3.84×109/L;hemoglobin:105 g/L;platelet count:174×109/L;and negativity for tumor markers.Abdominal enhanced computed tomography showed a space occupying lesion in the colon(1.9 cm).Fibrocolonoscopy revealed a polypoid and ulcerated mass measuring 2.5 cm.The tumor was removed.To our surprise,MS was confirmed by immunohistochemistry.PML/RARαfusion gene was detected in colon specimens by fluorescent in situ hybridization and real-time reverse transcription polymerase chain reaction,which was consistent with the bone marrow.She was diagnosed as having APL related MS.A smooth and unobstructed intestinal wall was found by fibrocolonoscopy,and continuous molecular remission was confirmed in both the bone marrow and colon after four courses of ATRA+arsenic trioxide(ATO).ATRA+ATO showed a favorable therapeutic response for both APL and MS.CONCLUSION Early use of ATRA can benefit APL patients,regardless of whether MS is the first or recurrent manifestation.

Gemtuzumab ozogamicin in the treatment of adult acute myeloid leukemia

Gemtuzumab ozogamicin (GO) is a humanized anti-CD33 monoclonal antibody conjugated to a derivative of an antitumor antibiotic, calicheamicin. GO was approved for the treatment of relapsed acute myeloid leukemia (AML) in the United States (US) in 2000. However, GO was withdrawn from the US market in June 2010, because a large-scale clinical trial failed to show additive or synergistic effects with conventional chemotherapy for newly diagnosed AML. GO is currently available only in Japan. However, several large clinical studies have demonstrated beneficial effects of GO when added to chemotherapy for AML in recent years;therefore, reconsideration of GO availability is gaining attention. Therefore, the role and efficacy of GO as monotherapy or in combination therapy for de novo or relapsed AML should be positively investigated.

Predictive Value of Dynamic Peri-Transplantation MRD Assessed By MFC Either Alone or in Combination with Other Variables for Outcomes of Patients with T-Cell Acute Lymphoblastic Leukemia

We performed a retrospective analysis to investigate dynamic peri-hematopoieticstem cell transplantation(HSCT)minimal/measurable residual disease(MRD)on outcomes inpatients with T-cell acute lymphoblastic leukemia(T-ALL).A total of 271 patients were enrolledand classified into three groups:unchanged ncgative MRD pre-and post-HSCT group(group A),post-MRD non-increase group(group B),and post-MRD increase group(group C).The patientsin group B and group C experienced a higher cumulative incidence of relapse(CIR)(42%vs.71%vs.16%,P<0.001)and lower leukemia-free survival(LFS)(46%vs.21%vs.70%,P<0.001)andoverall survival(OS)(50%vs.28%vs.72%,P<0.001)than in group A,but there was no significantdifference in non-relapse mortality(NRM)among three groups(14%vs.12%vs.8%,P=0.752).Multivariate analysis showed that dynamic peri-HSCT MRD was associated with CIR(HR=2.392,95%CI,1.816-3.151,P<0.001),LFS(HR=1.964,95%CI,1.546-2.496,P<0.001)and os(HR=1.731,95%CI,1.348-2.222,P<0.001).We also established a risk scoring system based ondynamic peri-HSCT MRD combined with remission status pre-HSCT and onsct of chronic graft-versus-host disease(GVHD).This risk scoring system could better distinguish ClR(c=0.730)thanthat for pre-HSCT MRD(c=0.562),post-HSCT MRD(c=0.616)and pre-and post-MRD dynamics(c=0.648).Our results confirm the outcome predictive value of dynamic peri-HSCT MRD eitheralone or in combination with other variables for patients with T-ALL.

PRAME Gene Expression in Acute Leukemia and Its Clinical Significance

Objective To investigate the expression of the preferentially expressed antigen of melanoma （PRAME） gene in acute leukemia and its clinical significance. Methods The level of expressed PRAME mRNA in bone marrow mononuclear cells from 34 patients with acute leukemia （AL） and in 12 bone marrow samples from healthy volunteers was measured via RT-PCR. Correlation analyses between PRAME gene expression and the clinical characteristics （gender, age, white blood count, immunophenotype of leukemia, percentage of blast cells, and karyotype） of the patients were performed. Results The PRAME gene was expressed in 38.2% of all 34 patients, in 40.7% of the patients with acute myelogenous leukemia （AML, n=27）, and in 28.6% of the patients with acute lymphoblastic leukemia （ALL, n=7）, but was not expressed in the healthy volunteers. The difference in the expression levels between AML and ALL patients was statistically significant. The rate of gene expression was 80% in M~, 33.3% in M2, and 28.6% in M~. Gene expression was also found to be correlated with CDl5 and CD33 expression and abnormal karyotype, but not with age, gender; white blood count or percentage of blast cells. Conclusions The PRAME gene is highly expressed in acute leukemia and could be a useful marker to monitor minimal residual disease. This gene is also a candidate target for the immunotherapy of acute leukemia.

Detection and clinical significance of multidrug resistance-1 mRNA in bone marrow cells in children with acute lymphoblastic leukemia by real-time fluorescence quantitative RT-PCR

Objective： Multidrug resistance（MDR） is one of the most important reasons for treatment failure and recurrence of acute leukemia. Its manifestations are different in children with acute lymphoblastic leukemia（ALL） which may be due to different detection methods. This study was to detect the expression of MDR1 mRNA in bone marrow cells of children with ALL by real-time fluorescence- quantitative reverse transcription polymerase-chain reaction（FQ-RT-PCR）, and combine minimal residual desease（MRD） detection by flow cytometry（FCM） and to study their relationship with treatment response and prognosis of ALL. Methods：The MDR1 mRNA levels in bone marrow cells from 67 children with ALL[28 had newly diagnosed disease, 27 had achieved complete remission（CR）, 12 recurrent] and 22 children without leukemia were detected by FQ-RT-PCR. MRD was detected by FCM. The patients were observed for 9-101 months, with a median of 64 months. Results：Standard curves of human MDR1 and GAPDH genes were constructed successfully. MDR1 mRNA was detected in all children with a positive rate of 100%. The mRNA level of MDR1 was similar among the newly diagnosed ALL group, CR group, and control group（P 〉 0.05）, but significantly higher in the recurrence group than that in newly diagnosed disease group and control group（0.50 ± 0.55 vs. 0.09 ± 0.26 and 0.12 ± 0.23, P〈 0.05）. 54 ALL patients were followed up, and it was found that MDR1 mRNA level was significantly higher in ALL patients within 3 years duration than that of ALL patients with 3-6 years and over 6 years duration（0.63 ± 0.56 vs. 0.11 ± 0.12 and 0.04 ± 0.06, P〈 0.01）. For the 28 children with newly diagnosed disease, the MDR1 mRNA level was similar between WBC 〉 50 ~ 109 group and WBC〈50 × 10^9 group（P〉 0.05）. In the 33 CR patients, the MDR1 mRNA level was significantly higher in MRD〉10a group than that in MRD〈10a group（0.39 ± 0.47 vs. 0.03 ± 0.03, P 〈 0.05）. Conclusion：The sensitivity and specificity of FQ-RT-PCR in detecting MDR1 mRNA in bone marrowy cells of children with ALL patients are high. MDR1 mRNA is expressed in children with and without leukemia. MDR1 mRNA is highly expressed in the CR ALL patients with high MRD, recurrence and short duration（within 3 years）. Monitoring MRD and the MDR1 mRNA level might be helpful for individual treatment.

Single-cell immune landscape of measurable residual disease in acute myeloid leukemia

Measurable residual disease(MRD)is a powerful prognostic factor of relapse in acute myeloid leukemia(AML).We applied the single-cell RNA sequencing to bone marrow(BM)samples from patients with(n=20)and without(n=12)MRD after allogeneic hematopoietic stem cell transplantation.A comprehensive immune landscape with 184,231 cells was created.Compared with CD8+T cells enriched in the MRDnegative group(MRD‒_CD8),those enriched in the MRD-positive group(MRD+_CD8)showed lower expression levels of cytotoxicity-related genes.Three monocyte clusters(i.e.,MRD+_M)and three B-cell clusters(i.e.,MRD+_B)were enriched in the MRD-positive group.Conversion from an MRD-positive state to an MRD-negative state was accompanied by an increase in MRD‒_CD8 clusters and vice versa.MRDenriched cell clusters employed the macrophage migration inhibitory factor pathway to regulate MRD‒_CD8 clusters.These findings revealed the characteristics of the immune cell landscape in MRD positivity,which will allow for a better understanding of the immune mechanisms for MRD conversion.

Effects of anti-CXCR_4 monoclonal antibody 12G5 on proliferation and apoptosis of human acute myelocytic leukemia cell line HL-60

Objective：To investigate the expression of CXCR4 on HL-60 cell line and the proliferation, apoptosis of HL-60 cell line cocultured with bone marrow stromal cells, so as to assess the possibility of 12G5, an anti-CXCR4 monoclonal antibody, in eradicating the minimal residual disease. Methods：The activity of SDF-1 was inhibited by 10 μg/ml 12G5. After treatment with 12G5, the status of adhesion was observed, and the adhesion rates, apoptosis and cell cycles were detected after 24 h of treatment. Cell growth rates were measured by trypan blue exclusion. Cell growth curve was plotted, and the expression of PCNA and apoptosis related protein including PCNA, Bcl-2 and Fas were detected with immunohistochemical technique. Results：（1） There was middling degree expression of CXCR4 on HL-B0 membrane. From 0 h to 6 h, as the time of 12G5 incubation along, the expression of CXCR4 decreased gradually. （2） After treatment for 24 h, the adhesion rates in the experiment group and the control were （39.4±7.9）% and （51.4±5.9）%, respectively. （3）After treatment for 24 h, the percentage of HL-60 cells in G0/G1 phase were （55.21±4.9）%, and that in S phase and G2/M phase were （30.40±4.1）% and （14.39± 5.2）%, respectively, with the corresponding proportions being （44. 67±2.2）%, （45.30±3.7）%, and （10. 03±2.6）% in the control. （4） The percentage of apoptotic HL-60 cells was （8.95±1.7）% in the experiment group, compared to （3. 97±2. 4）% in the control. （5）The survival rates of HL-60 cells decreased markedly at 48 h to 96 h, and the proliferation slowed down at this time duration. （6）The expression of PCNA and Bcl-2 down-regulated significantly, but the Fas protein expression was up-regulated. Conclusion：12G5 could inhibit the capability of adhesion and proliferation of HL-60 cells and it can induce more cells to enter G0/G1 phase and promote apoptosis. It may be helpful by inhibiting the bioactivity of SDF-1 with 12G5 in the therapy of marrow residual disease.

Current assessment and management of measurable residual disease in patients with acute lymphoblastic leukemia in the setting of CAR-T-cell therapy

Chimeric antigen receptor(CAR)-modified T-cell therapy has achieved remarkable success in the treatment of acute lymphoblastic leukemia(ALL).Measurable/minimal residual disease(MRD)monitoring plays a significant role in the prognostication and management of patients undergoing CAR-T-cell therapy.Common MRD detection methods include flow cytometry(FCM),polymerase chain reaction(PCR),and next-generation sequencing(NGS),and each method has advantages and limitations.It has been well documented that MRD positivity predicts a poor prognosis and even disease relapse.Thus,how to perform prognostic evaluations,stratify risk based on MRD status,and apply MRD monitoring to guide individual therapeutic decisions have important implications in clinical practice.This review assesses the common and novel MRD assessment methods.In addition,we emphasize the critical role of MRD as a prognostic biomarker and summarize the latest studies regarding MRD-directed combination therapy with CAR-T-cell therapy and allogeneic hematopoietic stem cell transplantation(allo-HSCT),as well as other therapeutic strategies to improve treatment effect.Furthermore,this review discusses current challenges and strategies for MRD detection in the setting of disease relapse after targeted therapy.

Outcomes of Adults with Acute Lymphoblastic Leukemia After Autologous Hematopoietic Stem Cell Transplantation and the Significance of Pretransplantation Minimal Residual Disease： Analysis from a Single Center of China

Background：The postremission therapics for adult patients generally contain consolidation chemotherapy,allogeneic hematopoietic stem cell transplantation and autologous hematopoietic stem cell transplantation （auto-HSCT）.Because of the various results from different centers,the optimal therapy for adult acute lymphoblastic leukemia （ALL） patients is still uncertain.This study aimed to better understand predictive factors and role of auto-HSCT in the postremission thcrapy for adult ALL patients.Methods：The outcomes of 135 adult patients with ALL,who received the first auto-HSCT in Hematopoietic Stem Cell Transplantation Center of Blood Diseases Hospital,Chinese Academy of Medical Sciences from January 1,1994 to February 28,2014,were retrospectively analyzed.Survival curves were estimated using the Kaplan-Meier method and simultaneous effects of multiple covariates were estimated with the Cox model.Results：Overall survival （OS） and disease-free survival （DFS） at 5 years for the whole cohort were 59.1 ± 4.5% and 59.0 ± 4.4%,respectively.The cumulative nonrelapse mortality and relapse rate at 5 years were 4.5 ± 0.03% and 36.6 ± 0.19%.For both OS and DFS,acute T-cell lymphoblastic leukemia,high lactate dehydrogenase （LDH） at diagnosis,blast cell proportion ≥5% on the 15th day of induction therapy,and extramedullary infiltration before HSCT were the poor prognosis factors.In addition,age ≥35 years predicted poor DFS.Only T-ALL and high LDH were the independent undesirable factors associated with OS and DFS in Cox regression model.For 44 patients who had results of pretransplantation minimal residual disease （MRD）,positive MRD （MRD ≥0.01%） indicated poor OS （P =0.044） and DFS （P =0.008）.Furthermore,for the standard risk group,the patients with negative MRD （MRD 〈0.01%） had better results （OS at 18 months was 90.0 ± 9.5%,while for the patients with positive MRD OS was 50.0 ± 35.4%,P =0.003;DFS at 18 months was 90.0 ± 9.5%,while for the positive MRD group DFS was 0%,P 〈 0.001）.Conclusions：This study confirmed that auto-HSCT combined with posttransplantation maintenance chemotherapy could be an option for adult ALL patients and pretransplantation MRD may play a significant role in the direction of therapy for adult ALL patients.

Quantitative monitoring of minimal residual disease in childhood acute lymphoblastic leukemia using TEL-AML1 fusion transcript as a marker

Importance: By demonstrating with TEL-AML1, this study indicated that mRNAs transcribed from fusion genes are ideal targets for minimal residual disease (MRD) monitoring in childhood acute lymphoblastic leukemia, and that different thresholds are needed to apply them into the risk stratification. Objective: TEL-AML1 expression was measured at three time points to 1) determine cut-off values for predicting acute lymphoblastic leukemia (ALL) relapse;2) investigate the prognostic value of this method and how well the results at these time points correlated;3) determine the correlation between MRD levels assessed using this marker and that determined by immunoglobulin/T-cell receptor (Ig/TCR) rearrangement detection. Methods: TEL-AML1 expression in 62 children with ALL was quantitated by real-time quantitative PCR at day 15, day 33, and month 3. The relationship between patient outcome and TEL-AML1 level was analyzed at each time point. The correlation between the MRD levels determined by TEL-AML1 or Ig/TCR rearrangements was also analyzed. results: For day 33, 6.68 TEL-AML1 copies/104 ABL copies was determined to be the best cut-off value. Higher levels were correlated with relapse (P = 0.001). For day 15 and month 3, the best cut-off values were 336.5 and 0.85 copies/104 ABL copies respectively;patients with higher expression levels had lower RFSs (day 15: P = 0.027;month 3:P = 0.023). For days 15 and 33, MRD levels assessed using TEL-AML1 or Ig/TCR rearrangements were strongly correlated [Spearman rank correlation coefficient (ρ) = 0.729 (day 15), 0.719 (day 33);P < 0.001 (both)], and both methods were equally effective at predicting relapse. At month 3, there was moderate correlation between the results derived from the two markers (ρ = 0.418, P = 0.003);however, receiver operating characteristic curve analysis showed that TEL-AML1 was a better prognostic marker. Interpretation: TEL-AML1 is an effective marker for MRD assessment and relapse prediction in children with ALL.

《Minimal residual disease and graft-versus-host disease guided multiple consolidation chemotherapy and donor lymphocyte infusion prevent second acute leukaemia relapse after allotransplant》解读--临床实践与基础研究的相互转化

本文最初发表于2016年《J Hematol Oncol》杂志上,文章题录为:Yan CH,Wang Y,Wang JZ,et al.Minimal residual disease and graft-versus-host disease guided multiple consolidation chemotherapy and donor lymphocyte infusion prevent second acute leukaemia relapse after allotransplant[J].J Hematol Oncol,2016,9:87。本研究证实,在异基因造血干细胞移植(allo-HSCT)后出现血液学复发的急性白血病、且经过诱导化疗+DLI达到完全缓解的患者中,MRD和GvHD指导下的多次巩固化疗+DLI可以预防再次复发,避免不必要的非复发死亡,最终改善生存。这项研究第一次强调了巩固化疗+DLI在治疗allo-HSCT后急性白血病复发中的作用。这将会成为治疗allo-HSCT后急性白血病复发的新方法。经通信作者许可,再次通过佳文解读的方式来阐述这一发现。

WT1基因指导的抢先治疗预防急性髓系白血病移植后复发的疗效及其最佳干预节点

目的:微量残留病(minimal residual disease,MRD)的监测与及时干预是预防成人急性髓系白血病(acute myeloid leukemia,AML)异基因造血干细胞移植(allogeneic hematopoietic stem cell transplantation,allo-HSCT)后复发的有效策略。泛白血病标志物WT1基因可作为AML患者MRD的监测指标。目前,临床上关于移植后基于WT1基因检测的抢先治疗的干预节点及干预时机尚无统一的标准。本研究旨在评估WT1基因指导的抢先治疗的临床价值,并进一步探讨其最佳干预节点。方法:回顾性收集2014年1月至2020年6月间在中南大学湘雅医院血液科接受alloHSCT的细胞遗传学风险为中危和高危成人AML患者的资料,所有患者均有移植后3年内的WT1基因表达检测数据。比较移植后接受抢先治疗的WT1基因阳性患者、未接受抢先治疗的WT1基因阳性患者之间,以及二者分别与移植后WT1基因阴性患者间终点指标[累积复发率(cumulative incidence of relapse,CIR)、无病生存(disease-free survival,DFS)率、总生存(overall survival,OS)率、非复发死亡(non-relapse mortality,NRM)率]的差异。纳入移植后未行干预的患者的数据,对可能影响预后的因素进行分析。将患者的年龄、性别、移植类型、细胞遗传学危险分层、移植前疾病状态、移植前WT1基因表达情况、移植后WT1基因表达情况、供者性别纳入单因素分析;进一步对单因素分析中P<0.10的因素纳入Cox回归模型进行多因素分析。采用受试者操作特征(receiver operating characteristic,ROC)曲线分析WT1基因表达水平预测复发的最佳截断值。结果:共纳入165例AML患者,86例在移植后3年内出现WT1基因阳性,其中58例接受抢先治疗,28例未接受抢先治疗。与移植后WT1基因阴性患者相比,未接受抢先治疗的WT1基因阳性患者的5年CIR明显升高(42.9%vs 10.5%,P<0.001)、5年DFS率(50.0%vs 80.7%,P=0.001)与OS率(60.7%vs82.8%,P=0.018)明显降低,5年NRM率差异无统计学意义(7.1%vs 8.9%,P=0.744);而接受抢先治疗的WT1基因阳性患者的上述指标差异均无统计学意义(均P>0.05)。多因素分析结果显示移植后WT1基因阳性是AML患者移植后预后不良因素(CIR:HR=6.24,P=0.0001;DFS率:HR=2.77,P=0.0096)。ROC曲线分析结果显示3年内WT1基因表达预测移植后复发的曲线下面积为0.727(95%CI 0.582~0.873),最佳截断值为122拷贝,敏感度为60.0%,特异度为89.9%。结论:连续监测细胞遗传学中高危AML患者allo-HSC后3年内WT1基因的表达,并对出现WT1基因表达阳性的患者进行及时的抢先治疗,能有效减少该类患者移植后的复发,改善预后。以WT1基因表达水平120拷贝作为抢先治疗的节点可能更为精准和可靠。

AML化疗期间不同时间点流式细胞术MRD检测对预后的影响

目的:探讨AML化疗期间不同时间点流式细胞术微小残留病(MRD)检测对预后的影响。方法:回顾性分析2018年3月到2022年3月确诊并规范化疗的130例成人原发AML患者,用流式细胞术检测MRD,Kaplan-Meier曲线进行生存分析,log-rank检验进行差异性分析,Cox比例风险回归模型进行影响患者生存的单因素和多因素分析。竞争风险模型进行影响患者累计复发率(CIR)的分析,Fine-Gray进行差异性分析。结果:130例患者中,CR181例,CR226例,PR 14例,NR 9例。CR1组OS高于CR2、PR、NR组。CR2组OS高于PR组,但与NR组比较无统计学差异;PR组OS与NR组比较无统计学差异。CR1和CR2的107例患者根据流式细胞术检测的MRD分组,第一次诱导化疗后MRD~-和MRD~+组患者的4年预期RFS率分别为65.3%和27.9%,4年预期OS率分别为58.7%和41.4%,4年预计CIR率分别为34.7%和69.7%,2组比较差异均有统计学意义(χ^(2)=6.639,P=0.010;χ^(2)=6.131,P=0.013;χ^(2)=6.637,P=0.010);第二次化疗后MRD~-和MRD~+组患者的4年预期RFS率分别为50.8%和37.9%,4年预期OS率分别为49.2%和44.5%,4年预计CIR率分别为49.2%和59.5%,2组比较差异均无统计学意义(χ^(2)=1.475,P=0.225;χ^(2)=2.432,P=0.119;χ^(2)=1.416,P=0.234);巩固治疗期间MRD~-和MRD~+组患者的4年预期RFS率分别为51.9%和29.6%,4年预期OS率分别为67.5%和24.6%,4年预计CIR率分别为48.1%和70.4%,2组比较差异均有统计学意义(χ^(2)=20.982,P<0.001;χ^(2)=17.794,P<0.001;χ^(2)=19.879,P<0.001);3个时间点MRD均为阴性和任一时间点为阳性患者的4年预期RFS率分别为69.9%和33.3%,4年预期OS率分别为59.1%和44.7%,4年预计CIR率分别为30.1%和65.1%,2组比较差异均有统计学意义(χ^(2)=7.367,P=0.007;χ^(2)=6.042,P=0.014;χ^(2)=7.662,P=0.006)。单因素分析结果显示,染色体高危核型是影响患者RFS和OS的不利因素,诱导化疗2个疗程达CR、第一次诱导化疗后MRD~-和第二次化疗MRD~-是患者RFS和OS的保护因素;巩固治疗期间MRD~-和3个时间点MRD~-是患者RFS、OS和CIR的保护因素。多因素分析结果显示,诱导化疗2个疗程达CR是患者RFS和CIR的保护因素,巩固治疗期间MRD~-是RFS、OS和CIR的保护因素。结论:成人AML患者早期获得CR和MRD~-,特别是巩固治疗期间的MRD~-是预后良好的标志,流式细胞术是AML患者MRD检测最常用的方法。

定期监测微小残留病变对儿童急性淋巴细胞白血病的预后价值

背景微小残留病变(MRD)用于监测和评估儿童急性淋巴细胞白血病(ALL)的治疗反应,并根据MRD水平进行危险度分层。目的探讨ALL患儿化疗期间及结束化疗后定期监测MRD对复发的预后价值。设计回顾性队列研究。方法收集2015年1月至2020年2月华中科技大学同济医学院附属同济医院(我院)接受CCCG-ALL 2015方案化疗的初诊ALL连续病例的临床资料,使用流式细胞术检测MRD。分析定期监测MRD与早期预测复发之间的关系。主要结局指标无复发生存期(RFS)。结果224例患儿纳入本文分析,男134例,女90例,中位年龄4.8岁。①诱导缓解第19天(D19)MRD阳性104例(46.4%),第46天(D46)MRD阳性23例(10.3%)。从诱导缓解后(16周)至结束化疗(125周)MRD均阴性145例。结束化疗后随访期间(152~287周),13例MRD阳性,其中11例(84.6%)复发。②28例患儿复发,中位复发时间33月,14例存活,12例死亡,2例失访;20例骨髓复发,其中2例合并睾丸复发,1例合并中枢神经系统复发(CNSL);8例单纯CNSL。③224例患儿随访时间52(IQR:36.5~69.5)月,5年RFS(84.5±2.8)%。D46 MRD≥0.01%与<0.01%、诱导缓解后至结束化疗期间MRD均阴性与MRD至少1次阳性患儿的5年RFS差异均有统计学意义(P<0.05)。结论D46 MRD≥0.01%及诱导缓解后至结束化疗期间MRD至少1次阳性的患儿预后较差。化疗过程中定期监测MRD十分重要。

儿童急性髓系白血病早期流式微小残留病检测与预后相关性的临床研究

目的:探索应用多参数流式细胞术(MFC)监测儿童急性髓系白血病(AML)诱导化疗后骨髓微小残留病(MRD)对预测预后的价值。方法:回顾性分析2015年8月-2022年12月武汉儿童医院血液科97例初诊AML患儿接受诱导化疗第Ⅰ周期和第Ⅱ周期后采用MFC检测骨髓MRD的结果,并分析其与预后的相关性。结果:97例AML患儿诱导治疗Ⅰ后57名患儿MRD阳性(MRD1^(+),58.8%),诱导治疗Ⅱ后仍有19名患儿MRD阳性(MRD2^(+),19.6%)。Kaplan-Meier生存分析结果显示37例(64.9%)MRD1^(+)接受移植治疗患儿预计3年总生存(OS)率明显高于20例(35.1%)未接受移植治疗患儿(84.6%vs 40.0%,P=0.0001)。35例MRD1^(+)MRD2^(-)患儿中,接受移植的25例患儿3年OS率高于10例未移植患儿(87.2%vs 70.0%,P=0.3229)。19例MRD1^(+)MRD2^(+)患儿3年OS率低于35例MRD1^(+)MRD2^(-)患儿(57.4%vs 81.8%,P=0.059)。12例MRD2^(+)移植治疗患儿3年OS率高于MRD2+7例未移植患儿(80.8%vs 14.3%,P=0.0007)。12例MRD1^(+)MRD2^(+)移植患儿和25例MRD1^(+)MRD2-移植患儿3年OS差异无统计学意义(80.8%vs 87.2%,P=0.8868)。7例MRD1^(+)MRD2^(+)未移植患儿3年OS明显低于10例MRD1^(+)MRD2^(-)未移植患儿(14.3%vs 70.7%,P=0.0114)。进一步多因素分析结果显示MRD2^(+)和移植治疗是患儿预后的独立影响因素(P=0.031,0.000),MRD1^(+)对预测97例患儿总体生存无统计学差异(P=0.902)。结论:诱导化疗Ⅱ后MRD阳性是AML患儿预后不良的独立危险因素。MRD2^(+)预示着更差的预后,可筛选出需接受移植的患儿。MRD2^(+)不是患儿移植治疗禁忌症尽早移植明显改善高危患儿的预后。