Association of the GSTP1 gene (Ile105Val) Polymorphism withChronic Myeloid Leukemia

The GSTP1 enzyme plays a key role in biotransformation and bioactivation of certain environmentalpollutants such as benzo[a]pyrene-7, 8-diol-9,10-epoxide (BPDE) and other diol epoxides of polycyclic aromatichydrocarbons. It catalyses the detoxification of base propanols that arise from DNA oxidation thus offering cellularprotection against oxidative stress. A single nucleotide polymorphism at codon 105 results in the substitutionof isoleucine (Ile) to valine (Val) causing a metabolically less active variant of the enzyme. We here assessed theimpact of the GSTP1 codon 105 polymorphism in chronic myeloid leukemia (CML) development and therapyresponse. The Ile105Val polymorphism was analyzed using a PCR-RFLP technique. Two hundred and sixtypatients with CML and 248 healthy, age and sex matched controls were included in the study of associations withpatient characteristics and treatment outcome. The GSTP1 Ile105Val polymorphism was significantly associatedwith CML development (χ2 = 9.57; df = 2; p = 0.0084). With respect to clinical phase, CML patients in advancedphase (accelerated and blast crisis) had higher frequency of heterozygous (Ile/Val) genotype (47.62%) comparedto chronic phase (36.5%). Further 54.5% of patients in blast crisis carried valine allele as compared to thosein chronic phase (36.5%). The frequency of combined genotypes (Ile/Val, Val/Val) was elevated in cytogeneticpoor (41.6%) and minor (53.57%) responders as compared to major (38.51%) responders. Hence the presentstudy suggests that GSTP1 Ile105Val polymorphism with reduced GSTP1 enzyme activity might influence CMLdevelopment, progression and response rates.     

Association of CYP3A5*3 and CYP1A1*2C Polymorphism with Development of Acute Myeloid Leukemia in Egyptian Patients

Aim: Cytochrome P450 (CYP) enzyme catalyzes the phase I metabolism reaction which metabolize endogenous and exogenous DNA-reactive chemical compounds and xenobiotics which could induce genotoxicity and increase the risk for leukemia. We aimed to detect frequency of CYP3A5*3 and CYP1A1*2C polymorphisms in Egyptian acute myeloid leukemia (AML) patients and to determine role of allele’s variants as a risk factor for developing leukemia. Patients and Methods: A case-control study was conducted on seventy acute myeloid leukemia patients and thirty control subjects. Samples were analyzed for prevalence of CYP3A5*3 and CYP1A1*2C polymorphisms using PCR - restriction fragment length polymorphism method. Results: CYP3A5*3 polymorphism (3/3) and (1/3) genotype were significantly elevated in AML group compared to control group (p=0.002). However, no statistical significant differences were found between patients and control group as regard CYP1A1*2C polymorphism. Conclusion: Our results suggest that Egyptians carrying CYP3A5*3 polymorphism might have an increased risk of AML emphasizing the significance of effective phase I detoxification in carcinogenesis.     

Cytogenetic Investigation in Chronic Myeloid Leukemia: Study from an Indian Population

Chronic myeloid leukemia (CML) is a malignant neoplasm of hematopoietic cells characterized by abnormalproliferation of myeloid precursors, decreased rates of self destruction and an arrest in cellular differentiation. Thebone marrow and peripheral blood accumulates all forms of mature and immature granulocytes, primarily blastcells. It is the most common type of leukemia seen in India, accounting for 30% of all leukemias. Cytogenetic analysisplays a vital and important role in the diagnosis of CML patients. The present study consists of cytogenetic evaluationof 175 CML cases from the Indian population with ages ranging from 6 – 86 years (mean of 42.8). The study populationincluded 115 males (65.72%) and 60 females (34.28%) with a Male: Female ratio 1.9:1. Out of the175 cases, 164(93.7%) were successfully karyotyped while culture failure was observed for 11 (6.3%). Among the 164 reportedcases, 53 (32.3%) showed a normal karyotype while within the 111 (67.7%) abnormal cases, 96 cases (86.5%) showedthe presence of Philadelphia (Ph’) chromosome with standard translocation t(9;22); Ph’+ve along with secondaryaberrations was detected in 9 (8.1%) cases. Variants of Ph’ chromosome were detected in only one case (0.9%). Ph’-ve CML with other chromosomal aberrations were detected in 5 (4.5%) cases, including +8, del 20q, del 11q andmarker chromosome. Furthermore, we believe that availability of more advanced molecular techniques can be usedas a supportive tool in CML diagnosis even though it cannot fully replace cytogenetics, which remains the backbonefor laboratory investigation of the disease.     

Detection of BCR/ABL Fusion Gene by Hematological and Cytogenetical Analysis in Chronic Myeloid Leukemia Patients in Quetta,Pakistan

Background: Chronic myeloid leukemia (CML) is a myeloproliferative disorder of pluripotent stem cells,caused by reciprocal translocation between the long arms of chromosomes 9 and 22, t(9;22)(q34;q11), known asthe Philadelphia chromosome. Materials and Methods: A total of 51 CML patients were recruited in this study.Complete blood counts of all CML patients were performed to find out their total leukocytes, hemoglobin andplatelets. FISH was performed for the detection of BCR-ABL fusion and cryptogenic tests using bone marrowsamples were performed for the conformation of Ph (9;22)(q34;q11) and variant translocation mechanisms.Results: In cytogenetic analysis we observed that out of 51 CML patients 40 (88.9%) were Ph positive and 4(8.88%) had Ph negative chromosomes. Mean values of WBC 134.5 103/μl, hemoglobin 10.44 mg/dl, and platelets288.6 103/μl were observed in this study. Conclusions: In this study, Ph positive translocation between chromosome(9:22)(q34;q11) were observed in 40 (88.9%) CML patients.     

Chronic Myeloid Leukemia - Prognostic Value of Mutations

Chronic myeloid leukemia (CML) is a stem cell disorder characterized by unrestricted proliferation of the yeloid series that occurs due to the BCR-ABL fusion oncogene as a result of reciprocal translocation t(9;22) q34;q11). This discovery has made this particular domain a target for future efforts to cure CML. Imatinib evolutionized the treatment options for CML and gave encouraging results both in case of safety as well as lerability profile as compared to agents such as hydroxyurea or busulfan given before Imatinib. However, about 2-4% of patients show resistance and mutations have been found to be one of the reasons for its development. European Leukemianet gives recommendations for BCR-ABL mutational analysis along with other tyrosine kinase inhibitors (TKIs) that should be administered according to the mutations harbored in a patient. The following overview gives recommendations for monitoring patients on the basis of their mutational status.     

CYP3A5基因多态性与晚期非小细胞肺癌紫杉醇化疗敏感性关系的研究

[目的]探讨CYP3A5基因多态性与晚期非小细胞肺癌紫杉醇化疗敏感性的关系。[方法]对接受以紫杉醇为基础化疗方案进行化疗的60例晚期非小细胞肺癌患者,采用PCR-RFLP方法检测外周血CYP3A5＊1/＊3基因多态性,比较不同基因型之间紫杉醇化疗近期疗效的差异。[结果]60例患者中,CYP3A5基因为＊1/＊1、＊1/＊3和＊3/＊3型的患者数分别有6例（10.00%）、20例（33.33%）和34例（56.67%）。＊1/＊1和＊1/＊3基因型患者的有效率为19.23%,＊3/＊3基因型患者的有效率为55.88%,＊1/＊1和＊1/＊3基因型患者的有效率明显低于＊3/＊3基因型患者（P=0.004）。[结论]对非小细胞肺癌患者治疗前进行CYP3A5基因型的测定,可预测紫杉醇的化疗疗效。     

Treatment and Survival in Patients with Chronic Myeloid Leukemia in a Chronic Phase in the West of Iran

Background: CML includes 30% of all leukemias, and occurs from childhood to old age. The present study was a retrospective analysis of chronic phase CML patients registered to a Hematology Clinic in Kermanshah, Iran, with checking of treatment options. Materials and Methods: Between 2002 and 2014, 85 CML patients referred to our hematology clinic were enrolled in our study. We surveyed age, sex, B-symptoms, splenomegaly, Sokal score, Hasford score, treatment and survival in all patients. Philadelphia chromosome analysis was conducted for each patient by conventional cytogenetics. We compared treatment in the patients with three drugs, imatinib, hydroxyurea (HU) and interferon alpha (IFN-α). Results: The mean age of the patients at diagnosis was 47.5±14.5 years (range, 23-82 years), with 43 (50.6%) being male. Some 13 (15.3%) were referred to our clinic for the first time with B-symptoms and 44 patients (51.8%) had splenomegaly. The Sokal score for 77 (90.6%) was low, 4 (4.7%) was intermediate and 4(4.7%) was high, but Hasford (Euro) scores for all patients were low. The 5-year survival rate for treated patients with imatinib, imatinib plus HU and imatinib plus HU plus IFN-α was 90.5%, 81.1% and 55.6%, respectively Conclusions: The results show that imatinib therapy alone provides better survival in CML patients compared to HU or IFN-α. Combinations of IFN-α and/or HU with imatinib probably reduce survival.     

Prevalence of BCR-ABL T315I Mutation in Malaysian Patients with Imatinib-Resistant Chronic Myeloid Leukemia

Objective: Chronic Myeloid Leukemia (CML) is caused by a reciprocal translocation between chromosomes 9and 22, t(9;22) (q34;q11) which encodes for the BCR-ABL fusion protein. Discovery of Imatinib Mesylate (IM) asfirst line therapy has brought tremendous improvement in the management of CML. However, emergence of pointmutations within the BCR-ABL gene particularly T315I mutation, affects a common BCR-ABL kinase contact residuewhich impairs drug binding thus contribute to treatment resistance. This study aims to investigate the BCR-ABL T315Imutation in Malaysian patients with CML. Methods: A total of 285 patients diagnosed with CML were included in thisstudy. Mutation detection was performed using qualitative real-time PCR (qPCR). Results: Fifteen out of 285 samples(5.26%) were positive for T315I mutations after amplification with real-time PCR assay. From the total number ofpositive samples, six patients were in accelerated phase (AP), four in chronic phase (CP) and five in blast crisis (BC).Conclusion: Mutation testing is recommended for choosing various tyrosine kinase inhibitors (TKIs) to optimizeoutcomes for both cases of treatment failure or suboptimal response to imatinib. Therefore, detection of T315I mutationin CML patients are clinically useful in the selection of appropriate treatment strategies to prevent disease progression.     

Prediction of Sensitivity to STI571 among Chronic Myeloid Leukemia Patients by Genome-wide cDNA Microarray Analysis

One of the most critical issues to be solved in regard to cancer chemotherapy is the establishment of ways to predict the efficacy of anti-cancer drugs for individual patients. To develop a prediction system based on expression of specific genes, we analyzed expression profiles of mononuclear cells from 18 chronic myeloid leukemia (CML) patients who were treated with the tyrosine kinase inhibitor STI571. cDNA microarrays representing 23 040 genes identified 79 genes that were expressed differentially between responders and non-responders to STI571. On the basis of the expression patterns of 15 or 30 of these genes among the patients, we developed a "Prediction Score" system that could clearly separate the responder group from the non-responder group. Verification of this system using four additional ("test") cases succeeded in predicting the response of each of those four patients to the drug. These results provide the first evidence that gene-expression profiles can predict sensitivity of CML cells to STI571, and may eventually lead to the achievement of "personalized therapy" for this disease.     

Immunophenotype and Ultrastructural Studies in Blast Crisis of Chronic Myeloid Leukemia

Thirty-four patients with chronic myeloid leukemia in blast crisis (CML-BC) were evaluated for lineage differentiation, with immunological markers and the presence of ultrastructural peroxidase. Eighteen (52.9%) were found to have myeloid blast crisis. Cytochemically, myeloperoxidase (MPO) could bedetectedonly in six patientson light microscopy while in the remaining 12 patients, myeloid differentiation was confirmed only by demonstration of MPO either at ultrastructural level or by the reactivity with anti myeloperoxidase (anti MPO) antibody. Six (17.6%) had lymphoid blast crisis as identified by lymphoid specific markers (CD 19; CD 10; CD7; CD4) along with the absence of myeloid markers. Heterogenous blast cell populations with mixed lineage differentiation were seen in 4 (I I .7%) patients. These cases showed both lymphoid (CD 19, CD 10) and myeloid (anti MPO and ultrastructural MPO) characteristics. A single case of megakaryoblastic blast crisis was identitied with positivity for CD41 and CD42 along with the presence of platelet peroxidase at the ultrastructural level. Five cases (148) of CML blast crisis remained unclassifiable. These results suggest that blast crisis in CML show an arrest of differentiation at an early stage when compared to de novo acute leukemias. This is particularly evident from the fact that MPO could only be demonstrated ultra-StNCtIIrally or with anti MPO antibody in the majority of patients with myeloid differentiation. It is expected that utilisation of molecular studies including immunoglobulin and T-cell receptor gene rearrangement and m-RNA expression for myeloperoxidase will provide a better insight into the level of differentiation for the presently unclassifiable cases of CML-blast crisis.     

Prognostic Relevance of DNMT3A,FLT3 and NPM1 Mutations in Syrian Acute Myeloid Leukemia Patients

Objective: Among all types of hematological neoplasms, acute myeloid leukemia (AML) has the highest death rate. Recently, cytogenetic and molecular genetics are crucial in the management, as a consequence of their effect on AML pathogenesis, classification, risk-stratification, prognosis and treatment. Methods: 100 Syrian adults with Normal Karyotype (NK) newly diagnosed  AML patients were included in this study, all cases confirmed histologically and immunohistochemically. Patients were divided into six subgroups using flow cytometry and cytological results. Polymerase chain reaction (PCR) was performed on exon 11-12 for FMS-like tyrosine kinase-3 internal tandem duplication (FLT3-ITD), exon 12 for Nucleophosmin1 (NPM1), and exon 23 for DNA methyltransferase 3A (DNMT3A) using target primers, the electropherograms were analyzed for gene mutations by comparing with the reference DNA sequence. Data were compared and aligned with different sequences using the NCBI BLAST Assembled Genomes tool. Results: FLT3-ITD, NPM1 and DNMT3A were detected in 24%, 22 % and 4%  patients respectively. M2 subtype had the most frequent incidence of diagnosis in AML. FLT3-ITD mutation patients had the highest mean of death cases, while the DNMT3A mutation patients had the lowest. On the other hand, the highest mean of remission was in patients with NPM1 mutation and the lowest in the carriers of the FLT3-ITD mutation. It was observed that the mean relapsed patients with FLT3-ITD and DNMT3A mutation was 3.4 and 2 months respectively, with no significant differences between (FLT3-ITD and DNMT3A) carriers and non-carriers relapsed. On the contrary,  the mean relapsed for NPM1 mutation carriers was 2.4  months with significant statistical differences. The mean survival time for patients with FLT3-ITD and NPM1  mutation was 5.9 months and 5.85 months respectively, with significant correlation. Between it was 5.88 months in DNMT3A patients with no significant differences. Finally, It was noted that the mean event free survival (EFS) of FLT3-ITD mutation patients was 4.818 months and the mean EFS of NPM1 mutation patients was 4.805 months, with significant statistical differences (p<0.05) between the mutation patients and non-mutated patients regarding to EFS, While this mean was not statistically significant in patients carrying DNMT3A mutation. Conclusion: Patients with FLT3-ITD and NPM1 mutations have the worst prognosis, where the presence of those mutations was significantly related to overall survival (OS) and EFS. Our study reflects that DNMT3A was not an extremely bad prognostic effect as an independent factor. We can declare according to this study that genetic mutation and variants detection could easily be incorporated into the regimen evaluation of AML patients.     

Cardiovascular Risk and Cardiovascular Events in Patients With Chronic Myeloid Leukemia Treated With Tyrosine Kinase Inhibitors

Background

Cardiovascular events (CVEs) have been observed in patients with chronic myeloid leukemia treated with second-generation tyrosine kinase inhibitors.

Association of Multidrug Resistance Gene-1 (MDR1 C1236T) Polymorphism with the Risk of Acute Myeloid Leukemia in a Moroccan Population

The human multidrug resistance MDR1 gene plays a crucial role in the absorption, transport, metabolism and elimination of harmful compounds. An impaired metabolism of these compounds related to genetic polymorphism may cause cancer such as acute myeloid leukemia AML. Objective: The present study investigated the relationship between C1236T polymorphism and the risk of AML development in a sample of Moroccan population. Methods: The present case-control study included 131 AML patients and 136 healthy controls. The MDR1 C1236T polymorphism was identified by PCR-RFLP method. Meta-analysis was performed to discuss our results. Statistical analyses were performed using SPSS, MetaGenyo and MedCalc. Results: A positive association was found between the 1236TT mutant genotype and the risk of AML (OR 2.39; 95% CI 1.02-5.57, p= 0.04) compared to the wild type 1236CC. In addition, the recessive model revealed that carriers of 1236TT mutant genotype were more exposed to develop AML when compared to the combined 1236CC/CT genotype (OR: 2.27, CI: 1.01–5.05, p=0.04). The clinical parameters of AML showed no significant association. Meta-analysis demonstrated no statistically significant association between this polymorphism and AML susceptibility. Conclusion: Our study suggests that the MDR1C1236T polymorphism appears to be associated with the risk of AML. Further studies, including a large sample size, are needed to confirm these findings.     

Heterogeneity of V52 TCR 8 Gene Rearrangements in Acute Myeloid Leukemia

In a previous study, we described the occurrence of T-cell receptor (TCR)δ gene rearrangements in 9/100 acute myeloid leukemia (AML) cases. In this study, we further characterized these rearrangements by Southern Blot hybridization using a Vδ2 specific probe and polymerase chain reaction (PCR). Southern Blot analysis revealed that rearrangements involved the Vδ2 gene segment in four patients. Interestingly the restriction fragments detected by the Vδ2 probe differed markedly in size. PCR analysis revealed a complete Vδ2(Dn)Jδ1 gene rearrangement, an incomplete Vδ2Dδ3 rearrangement and a large amplification product, which cannot be explained with normal VDJ recombinatorial processes, in one case each. Furthermore although Vδ2 and Jδ1 were rearranged, no comigration of rearranged fragments was observed and no PCR product was obtained in one case.

Obtained results in AML differ from findings in acute lymphoblastic leukemia (ALL) of B-cell lineage, where a more homogeneous pattern of rearrangements has been described. This heterogeneity might be related to the illegitimate occurrence of TCRδ gene rearrangements in AML.     

金克对慢性粒细胞性白血病细胞因子的影响

「目的」观察金克冲剂治疗慢性粒细胞性白血病（慢性期）的疗效,并探讨细胞因子水平的意义。「方法」收集３０例慢粒慢性期患者,用金克冲剂治疗,观察患者的临床疗效、细胞因子水平。「结果」治疗组完全缓解率３６．６７％,部分缓解率４３．３３％,总有效率８０％。治疗组细胞因子水平中白介素２,可溶性白介素２受体,肿瘤坏死因子α,干扰素γ治疗前后有显著差异;且治疗后与正常对照组比较无显著差异（Ｐ〉０．０５）。「结论」金克冲剂对慢性粒细胞性白血病慢性期疗效确切,其作用机理可能与改善细胞因子水平有关。     

Neuropilin-1基因在髓细胞白血病和正常人骨髓基质细胞中的表达

目的了解neuropilin-1(NP-1)基因在髓细胞白血病(AML和CML)患者及正常人骨髓基质细胞中的表达情况。方法收集12例AML、14例CML和20例正常对照骨髓标本,分离单个核细胞。进行体外长期培养,收集贴壁细胞(骨髓基质细胞)。利用逆转录-聚合酶链反应分别检测3组骨髓基质细胞中NP-1基因的表达。结果成功建立了AML、CML和正常人骨髓基质细胞培养方法,NP-1基因在AML、CML骨髓基质细胞中的表达率分别为47.1%和50%,明显低于正常对照(85%)。结论NP-1基因可表达于部分AML、CML和大部分正常人的骨髓基质细胞中,其在髓细胞白血病患者骨髓基质细胞中的低表达, 可能与其调节造血功能异常有关。     

Neuropilin-1某因在髓细胞白血病和正常人骨髓基质细胞中的表达(英文)

目的了解 neuropilin-1(NP-1)基因在髓细胞白血病(AML 和 CML)患者及正常人骨髓基质细胞中的表达情况。方法收集12例 AML、14例 CML 和20例正常对照骨髓标本,分离单个核细胞。进行体外长期培养,收集贴壁细胞(骨髓基质细胞)。利用逆转录-聚合酶链反应分别检测3组骨髓基质细胞中 NP-1基因的表达。结果成功建立了 AML、CML 和正常人骨髓基质细胞培养方法,NP-1基因在 AML、CML 骨髓基质细胞中的表达率分别为47.1%和50%,明显低于正常对照(85%)。结论 NP-1基因可表达于部分 AML、CML 和大部分正常人的骨髓基质细胞中,其在髓细胞白血病患者骨髓基质细胞中的低表达,可能与其调节造血功能异常有关。