Lymphoma of host origin in a marrow transplant recipient in remission of acute myeloid leukemia and receiving cyclosporin A

Lymphomas are an uncommon complication of solid organ transplantation and rarely occur after marrow transplantation. When post-marrow transplant lymphomas have occurred, they have been of donor cell origin and when sought, Epstein-Barr virus DNA has been found in the tumor. A 21-year-old woman developed a poorly differentiated lymphocytic lymphoma 6 months after bone marrow transplantation for acute myeloid leukemia in remission. Cyclosporin A had been used as an immunosuppressant. A chromosomal polymorphism demonstrated that the tumor was of host origin and contained a monoclonal tumor marker, 46,XX INV 4 (p16q12). The tumor did not contain the DNA of the Epstein-Barr virus.     

Cytogenetic changes and their prognostic significance in elderly acute myeloid leukemia

Cytogenetic analyses were performed in 72 acute myeloid leukemia (AML) patients > or = 60 years old. Karyotype was normal in 35 (48.6%) patients (group III). 3 patients (4.2%) had favourable karyotype with t(15;17) as an isolated aberration (group IV). 21 patients (29.2%) had adverse karyotypes (group I) and 13 (18%) had intermediate karyotypes (group II). Adverse karyotypes were simple (< 3 aberrations), with add 3q, 5q-, 7q-, in 5 persons, and complex (> or = 3 aberrations) in 16. Karyotypes of 14 patients from the latter group contained > or = 5 aberrations. Laboratory and clinical data were comparable between groups with > or = 3 and with > or = 5 changes. In more than 2/3 complex karyotypes chromosome 5 and 7 aberrations also were found. AML clinical course of group II patients was more similar to that of group I than of groups III and IV. A frequency of complete remissions differed statistically between group I and the others and a frequency of complete and partial remissions together--between I + II and III + IV groups. Overall survival time differed statistically between all groups. There were significantly more patients with secondary AML in groups I and II than in group III. Analysis according to FAB did not show prognostic significance of this classification. Authors conclude that cytogenetics have a fundamental prognostic importance in AML of the elderly and should be taken into account in establishing therapeutic strategies.     

α-干扰素治疗慢性髓细胞性白血病的骨髓组织学改变

目的 :探讨长期使用α 干扰素 (α INF)治疗对慢性髓细胞性白血病 (CML)患者骨髓组织的影响。方法 :应用常规HGF染色、CD4 1免疫组织化学染色及Gomori银浸染色方法 ,对比分析 2 9例CML患者经α INF联合羟基脲 (HU)或HU单剂治疗后的骨髓塑包切片上粒系、巨核系及网硬蛋白纤维的改变。结果 :长期使用α INF治疗后 ,造血组织及粒 /红细胞比例分别下降至 80 .4 %及 4 .0∶1,与对照组相比差异有统计学意义 (P <0 .0 5 ) ;同时 ,网状纤维含量增高 (P <0 .0 5 ) ,而CD4 1细胞数量无明显改变 ,其形态以成熟巨核细胞为主。结论 :长期使用α INF治疗对CML患者骨髓造血主质及造血间质均有影响 ,骨髓切片有助于全面评价α INF的治疗作用     

Characterization and prognostic significance of mitochondrial DNA variations in acute myeloid leukemia

Recent studies have suggested that mutations in the mitochondrial genome (mtDNA) may play a role in the development and response to treatment for human cancer. The aim of this study was to investigate whether mtDNA variations have any prognostic relevance, to clarify the spectra of mtDNA variation and to determine whether there was any correlation to known prognostic factors in acute myeloid leukemia (AML). To elucidate this, we sequenced the entire mtDNA in 56 AML patients and 14 control subjects. When analyzing the biologic impact of the non‐synonymous variations in the mtDNA coding genes, we found an inferior disease‐free survival for patients exhibiting variations in the two most important catalytic genes of the complex IV of the oxidative phosphorylation complexes (OXPHOS), that is, the cytochrome c oxidase subunit I and the cytochrome c oxidase subunit II (hazard ratio 2.6, P = 0.03; multivariate analysis). In addition, the most frequent variation was the T16311C in the control region, which was found in 11 (20%) of the 56 patients. This observation was confirmed in another cohort of 173 diagnostic AML samples. In this expanded group, the T16311C variation tended to be associated with chromosomal abnormalities.     

Clinical and histological features retain their prognostic impact under interferon therapy of CML: A pilot study

In 55 patients with Ph¹⁺ CML under interferon (IFN) monotherapy, an immunohistochemical and morphometric study on pretreatment bone marrow biopsies was performed to evaluate the prognostic impact of clinical as well as histological disease features. For identification of megakaryocytes we used the PAS stain and CD61 to calculate the subtraction of precursors (pro- and megakaryoblasts). Demonstration of macrophages and their different subsets was carried out by PG-M1 (CD68) and the GSA-I lectin. The erythroid precursors were stained by Ret40f (anti-glycophorin C). Density of argyrophilic (reticulin plus collagen) fibers was determined by applying Gomori's silver impregnation method. Clinical variables like state of hematological response to IFN administration, age, spleen and liver size, myeloblasts plus promyelocytes, basophils as well as basophils and eosinophils exerted a predictive capacity by univariate statistical analysis. However, when entering these factors into previously published risk models, i.e., the so-called Sokal score and its modifications, to assess subgroups with different survival patterns or relative risk groups, a clear-cut discrimination was not feasible. Bone marrow features of prognostic value consisted of megakaryocytes and their precursors, fibers, and pro- and erythroblasts. Only when including histological variables into a formerly reported Cox model, could a significant separation of patients into the different categories or relative risk groups be computated. In conclusion, the present data emphasize the prognostic impact of histological parameters to be considered in all clinical trials on CML.     

急性白血病骨髓中的血管生成的研究

目的 研究血管生成在急性白血病发病,预后中的作用。方法 应用免疫组化法检测30例初治急性白血病患者的骨髓活检组织治疗前后微血管密度（MVD）及血管内皮生长因子（VEGF）的变化。结果 初治急性白血病患者骨髓病理组织中的MVD和VEGF阳性率明显高于正常对照组;骨髓完全缓解后其MVD及VEGF阳性率明显下降,与正常对照组无显著性差异;治疗后未完全缓解组的MVD及VEGF阳性率较治疗前无显著性下降。结论 急性白血病骨髓中存在血管生成,可能与急性白血病的预后有关。     

Therapy for acute myeloid leukemia in 119 adults: A comparison of two treatment protocols

Summary Of 119 patients with acute myeloid leukemia, 69 were treated with Adriamycin, Vincristine and Cytosine Arabinoside (Therapy 1) and 50 with Daunorubicin, Cytosine Arabinoside and 6-Thioguanine (Therapy 2) as well as a consolidation therapy. The maintenance therapy with Cytosine Arabinoside and 6-Thioguanine was the same for both groups. The complete remission rate was 44% for Therapy 1 and 68% for Therapy 2 (p<0.05). — The median values for remission duration were 7 and 13 months respectively (p=0.10); for survival time the median values were 18 and 19 months. These figures show in retrospect that high remission rates can be attained through intensive induction therapy and that longer remission duration is correlated with more aggressive induction therapy. A mild form of maintenance therapy seems to have little effect on the duration of complete remission.     

Clinical significance of Treg cell frequency in acute myeloid leukemia

This study was designed to investigate the clinical significance of peripheral blood CD4⁺ CD25⁺ CD127 low-regulatory T (T_reg) cells in acute myeloid leukemia (AML) patients. T_reg cells in the peripheral blood of 80 AML patients and 35 age-matched healthy controls were counted by flow cytometry. Correlations between the frequency of circulating T_reg cells and disease status, treatment outcome, or prognosis were evaluated. The percentages of T_reg cells in patients at diagnosis and during refractory/relapse were significantly higher than that in healthy controls. There was no significant difference in the percentages of T_reg cells between patients in remission and healthy controls. After six cycles of chemotherapy, the percentage of T_reg cells in patients who achieved complete remission was significantly lower than that in patients at diagnosis, but there was no difference in T_reg frequency between refractory/relapse patients and patients at diagnosis. T_reg cells in the peripheral blood of AML patients may play a suppressive role in host antitumor immune response. The frequency of T_reg cells in peripheral blood may thus be used as a biomarker for predicting sensitivity to chemotherapy and prognosis of AML patients. Additionally, T_reg number in peripheral blood could be used to monitor disease status and evaluate disease progression.     

Clinical significance of genetic aberrations in secondary acute myeloid leukemia

The study aimed to identify genetic lesions associated with secondary acute myeloid leukemia (sAML) in comparison with AML arising de novo (dnAML) and assess their impact on patients' overall survival (OS). High‐resolution genotyping and loss of heterozygosity mapping was performed on DNA samples from 86 sAML and 117 dnAML patients, using Affymetrix Genome‐Wide Human SNP 6.0 arrays. Genes TP53, RUNX1, CBL, IDH1/2, NRAS, NPM1, and FLT3 were analyzed for mutations in all patients. We identified 36 recurrent cytogenetic aberrations (more than five events). Mutations in TP53, 9pUPD, and del7q (targeting CUX1 locus) were significantly associated with sAML, while NPM1 and FLT3 mutations associated with dnAML. Patients with sAML carrying TP53 mutations demonstrated lower 1‐year OS rate than those with wild‐type TP53 (14.3% ± 9.4% vs. 35.4% ± 7.2%; P = 0.002), while complex karyotype, del7q (CUX1) and del7p (IKZF1) showed no significant effect on OS. Multivariate analysis confirmed that mutant TP53 was the only independent adverse prognostic factor for OS in sAML (hazard ratio 2.67; 95% CI: 1.33–5.37; P = 0.006). Patients with dnAML and complex karyotype carried sAML‐associated defects (TP53 defects in 54.5%, deletions targeting FOXP1 and ETV6 loci in 45.4% of the cases). We identified several co‐occurring lesions associated with either sAML or dnAML diagnosis. Our data suggest that distinct genetic lesions drive leukemogenesis in sAML. High karyotype complexity of sAML patients does not influence OS. Somatic mutations in TP53 are the only independent adverse prognostic factor in sAML. Patients with dnAML and complex karyotype show genetic features associated with sAML and myeloproliferative neoplasms. Am. J. Hematol., 2012. © 2012 Wiley Periodicals, Inc.     

Gefitinib induces myeloid differentiation of acute myeloid leukemia

Cure rates for patients with acute myeloid leukemia (AML) remain low despite ever-increasing dose intensity of cytotoxic therapy. In an effort to identify novel approaches to AML therapy, we recently reported a new method of chemical screening based on the modulation of a gene expression signature of interest. We applied this approach to the discovery of AML-differentiation-promoting compounds. Among the compounds inducing neutrophilic differentiation was DAPH1 (4,5-dianilinophthalimide), previously reported to inhibit epidermal growth factor receptor (EGFR) kinase activity. Here we report that the Food and Drug Administration (FDA)-approved EGFR inhibitor gefitinib similarly promotes the differentiation of AML cell lines and primary patient-derived AML blasts in vitro. Gefitinib induced differentiation based on morphologic assessment, nitro-blue tetrazolium reduction, cell-surface markers, genome-wide patterns of gene expression, and inhibition of proliferation at clinically achievable doses. Importantly, EGFR expression was not detected in AML cells, indicating that gefitinib functions through a previously unrecognized EGFR-independent mechanism. These studies indicate that clinical trials testing the efficacy of gefitinib in patients with AML are warranted.     

Clinical characteristics and prognostic implications of NPM1 mutations in acute myeloid leukemia

Recently, somatic mutations of the nucleophosmin gene (NPM1), which alter the subcellular localization of the product, have been reported in acute myeloid leukemia (AML). We analyzed the clinical significance of NPM1 mutations in comparison with cytogenetics, FLT3, NRAS, and TP53 mutations, and a partial tandem duplication of the MLL gene (MLL-TD) in 257 patients with AML. We found NPM1 mutations, including 4 novel sequence variants, in 64 of 257 (24.9%) patients. NPM1 mutations were associated with normal karyotype and with internal tandem duplication (ITD) and D835 mutations in FLT3, but not with other mutations. In 190 patients without the M3 French-American-British (FAB) subtype who were treated with the protocol of the Japan Adult Leukemia Study Group, multivariate analyses showed that the NPM1 mutation was a favorable factor for achieving complete remission but was associated with a high relapse rate. Sequential analysis using 39 paired samples obtained at diagnosis and relapse showed that NPM1 mutations were lost at relapse in 2 of the 17 patients who had NPM1 mutations at diagnosis. These results suggest that the NPM1 mutation is not necessarily an early event during leukemogenesis or that leukemia clones with NPM1 mutations are sensitive to chemotherapy.     

Clinical significance of minimal residual disease in childhood acute myeloid leukemia

Many studies have assessed the clinical significance of the detection of minimal residual disease (MRD) in acute leukemia. Thus far, many studies have suggested that MRD detection to evaluate the response to chemotherapy is useful for predicting the prognosis of childhood acute lymphoblastic leukemia (ALL). However, few studies have reported on the significance of MRD in childhood acute myeloid leukemia (AML), because of small numbers of patients and limited availability of MRD markers. Therefore, we monitored MRD using currently available markers at several points during the treatment for childhood AML and tried to intensify the treatment based on the results of MRD. Thirty-one patients (26 de novo cases and 5 other cases) were examined for MRD between February 1999 and May 2002. After the first consolidation therapy (consolidation 1), the expression of Wilms tumor gene (WT1) and/or leukemia-specific fusion genes such as AML1/MTG8, PML/RAR alpha, and MYH11/CBF beta were analyzed. Patients with positive MRD but in hematological remission at that point were recommended to undergo stem cell transplantation (SCT). Positive WT1 expression (more than 10(3) copies/microgram RNA) was detected in 18 of 31 patients (58.1%) at onset. After consolidation 1 therapy, the WT1 expression became negative in 14 of 18 patients. The AML1/MTG8 fusion gene was expressed in 8 patients, PML/RAR alpha was expressed in 3 patients, and MYH11/CBF beta was expressed in 1 patient. Four of the 8 patients with AML1/MTG8 expression and all 3 with PML/RAR alpha expression also demonstrated positive WT1 expression at onset. Eight (5 de novo cases and 3 other cases) of the 31 patients had no available MRD markers. Four patients who showed pesistently high expression of WT1 after consolidation 1 therapy underwent SCT, and only 1 patient remained in complete remission (CR). Among 14 patients who became negative for WT1 expression, 6 patients received SCT for various reasons. Among 8 patients with the AML1/MTG8 fusion gene, 2 became MRD negative and 6 continued to be positive. Four of these 6 patients underwent SCT, and all but one who underwent syngeneic SCT became MRD negative. On the other hand, 1 of the 2 patients who continued on chemotherapy continued to be MRD positive, suggesting a graft-versus-leukemia effect in allogeneic SCT. All patients with the PML/RAR alpha and MYH11/CBF beta fusion gene continued to be in CR. The 3-year event-free survival in de novo AML was 69.4% +/- 9.8% (n = 26), a result that is encouraging and superior to other reported outcomes. Thus, an MRD-based treatment strategy together with conventional risk factors appears to be required for further improving the outcomes of AML.     

Clinical significance of RAS pathway alterations in pediatric acute myeloid leukemia

RAS pathway alterations have been implicated in the pathogenesis of various hematological malignancies. However, their clinical relevance in pediatric acute myeloid leukemia (AML) is not well characterized. We analyzed the frequency, clinical significance, and prognostic relevance of RAS pathway alterations in 328 pediatric patients with de novo AML. RAS pathway alterations were detected in 80 (24.4%) of 328 patients: NF1 (n=7, 2.1%), PTPN11 (n=15, 4.6%), CBL (n=6, 1.8%), NRAS (n=44, 13.4%), KRAS (n=12, 3.7%). Most of these alterations in the RAS pathway were mutually exclusive also together with other aberrations of signal transduction pathways such as FLT3-ITD (P=0.001) and KIT mutation (P=0.004). NF1 alterations were frequently detected in patients with complex karyotype (P=0.031) and were found to be independent predictors of poor overall survival (OS) in multivariate analysis (P=0.007). At least four of seven patients with NF1 alterations had biallelic inactivation. NRAS mutations were frequently observed in patients with CBFB-MYH11 and were independent predictors of favorable outcomes in multivariate analysis (OS, P=0.023; event-free survival [EFS], P=0.037). Patients with PTPN11 mutations more frequently received stem cell transplantation (P=0.035) and showed poor EFS than patients without PTPN11 mutations (P=0.013). Detailed analysis of RAS pathway alterations may enable a more accurate prognostic stratification of pediatric AML and may provide novel therapeutic molecular targets related to this signal transduction pathway.     

Clinically useful prognostic factors in acute myeloid leukemia

The clinical outcome of acute myeloid leukemia (AML) is extremely variable, ranging from survival of a few days to cure. Different clinical and biological features at diagnosis have been reported as useful for the prediction of clinical outcome; however, in most AML cases induction therapy must be initiated as soon as possible, therefore, the possibility of stratifying patients at diagnosis is generally not taken into account, with the exception of acute promyelocytic leukemia in which morphology, immunophenotype, and molecular biology allow a rapid diagnosis and the adoption of specific therapy. As a consequence, prognostic factors in AML are more useful for the prediction of relapse, rather than for the stratification of induction therapy. Most relevant studies, based on large multicenter trials have definitively demonstrated that age and cytogenetics at diagnosis are the most important prognostic determinants for patients with AML. Early blast clearance after induction chemotherapy represents a further important factor of potential utility into clinical practice. Finally, biologic parameters, such as mutations of FLT3 and nucleophosmin, have been reported as useful for the prognostic categorization mainly in patients with intermediate cytogenetics and can also represent potential targets for new therapeutic agents.     

Prevalence and prognostic significance of Flt3 internal tandem duplication in pediatric acute myeloid leukemia

The Flt3 gene encodes a tyrosine kinase receptor that regulates proliferation and differentiation of hematopoietic stem cells. An internal tandem duplication of the Flt3 gene (Flt3/ITD) has been reported in acute myelogenous leukemia (AML) and may be associated with poor prognosis. We analyzed diagnostic bone marrow specimens from 91 pediatric patients with AML treated on Children's Cancer Group (CCG)-2891 for the presence of the Flt3/ITD and correlated its presence with clinical outcome. Fifteen of 91 samples (16.5%) were positive for the Flt3/ITD. Flt3/ITD-positive patients had a median diagnostic white count of 73 800 compared with 28 400 for the Flt3/ITD-negative patients (P =.05). The size of the duplication ranged from 21 to 174 base pairs (bp). Nucleotide sequencing of the abnormal polymerase chain reaction products demonstrated that all duplications involved exon 11 of the Flt3 gene and also preserved the reading frame. Lineage restriction analysis revealed that Flt3/ITD was not present in the lymphocytes, suggesting a lack of stem cell involvement for this mutation. None of the Flt3/ITD-positive patients had unfavorable cytogenetic markers, and there was no predominance of a particular FAB class. The remission induction rate was 40% in Flt3/ITD-positive patients compared with 74% in Flt3/ITD-negative ones (P =.005). The Kaplan-Meier estimates of event-free survival at 8 years for patients with and without Flt3/ITD were 7% and 44%, respectively (P =.002). Multivariate analysis demonstrated that presence of the Flt3/ITD was the single most significant, independent prognostic factor for poor outcome (P =.009) in pediatric AML.     

Acute myeloid leukemia of donor origin after allogeneic bone marrow transplantation for precursor T-cell acute lymphoblastic leukemia: case report and review of the literature

We report a case of donor-derived acute myeloid leukemia (AML) occurring in a 33-year-old man after allogeneic bone marrow transplantation (BMT) for precursor T-cell acute lymphoblastic -leukemia (T-ALL). The cells for BMT were from his human leukocyte antigen (HLA)-matched sister. Fluorescence in-situ hybridization (FISH) analysis showed the AML to be of donor origin (i.e., karyotypically female) with an 11q23 (mixed lineage leukemia (MLL) gene) translocation, while the original T-ALL exhibited a male karyotype with abnormalities of chromosomes 6, 8, and a t(10;14)(q24;q11.2). Subsequent molecular short tandem repeat studies confirmed the AML to be of donor origin. Donor-cell leukemia (DCL) after allogeneic BMT is a rare, yet well-documented, event. Our report presents clinicopathologic information about a case of DCL and a review of the recent literature.