Fluorescence In Situ Hybridization Monitoring of BCR-ABL-Positive Neutrophils in Chronic-Phase Chronic Myeloid Leukemia Patients during the Primary Stage of Imatinib Mesylate Therapy

We describe a method for monitoring chronic myeloid leukemia (CML) patients treated with imatinib that uses fluorescence in situ hybridization (FISH) to detect BCR-ABL in peripheral blood (PB) granulocytes. First, we compared this method, termed Neutrophil-FISH, with interphase FISH (i-FISH) analysis of bone marrow (BM), i-FISH analysis of PB mononuclear cells, and conventional cytogenetic analysis (CCA) of BM in 30 consecutive CML patients. We found the percentage of BCR-ABL-positive neutrophils as determined by Neutrophil-FISH to correlate best with the percentage of Philadelphia chromosome-positive metaphases in the BM determined by CCA (y = 0.8818x + 5.7249; r(2) = 0.968).We then performed a serial Neutrophil-FISH study of 10 chronic-phase CML patients treated with imatinib and found that the technique could clearly separate imatinib responders from nonresponders within 12 weeks of drug administration. There was a significant difference in the percentages of BCR-ABL-positive neutrophils between responder (mean 3 SD, 18.2% 3 11.8%) and nonresponder (82.4% 3 5.1%) groups at 12 weeks (P < .0001, Student t test).Together with real-time quantitative polymerase chain reaction analysis, Neutrophil-FISH represents another useful method for monitoring CML patients during the primary myelosuppressive stage of imatinib therapy because it is a quick, simple, and reliable method for assessing cytogenetic response.     

Short-term vasculoprotective effects of imatinib mesylate on intimal hyperplasia of arterial anastomosis: An experimental study using a rabbit model

BACKGROUND:

Since the beginning of the ‘microvascular era’, the success rates of microvascular procedures have increased to more than 90% in most series. The main reason for failure, however, is the healing of microarterial anastomosis, which is dependent on the status of endothelial cells and affects the rate of arterial thrombosis. In 80% of arterial thrombosis cases, complications are primarily observed during the first 72 h after surgery. Healing of arterial anastomosis results in intimal hyperplasia in which myofibroblasts comprise the predominant cell type. Intimal hyperplasia has been described previously as an adaptive process that occurs in response to hemodynamic stress or injuries to the vascular bed. During wound healing, fibroblasts proliferate, migrate and differentiate into myofibroblasts – a process that takes one to three days. Imatinib mesylate (ST1571-Gleevec, Novartis, Germany) is a specific platelet-derived growth factor receptor blocker that has found use as an adjunct to sirolimus in cardiovascular surgery for restenosis. However, its potential utility in preventing arterial thrombosis in microvascular surgery has not been evaluated in routine plastic surgery practice.

METHODS:

Twenty-four randomly selected, male, white New Zealand rabbits were divided into six groups (A to F), and the femoral artery model was used for arterial anastomosis. Following anastomosis, groups A, B and C received phosphate-buffered saline orogastrically. In groups D, E and F, imatinib mesylate was administered via an orogastric tube twice per day at a dose of 10 mg/kg starting two days before arterial anastomosis. Following anastomosis, imatinib mesylate was administered for one, three and seven days, and the regression of intimal hyperplasia was recorded.

RESULTS:

In groups administered imatinib mesylate (ie, groups D, E and F), intimal hyperplasia decreased by up to 50%, which represented a statistically significant difference. Histological analysis confirmed smooth muscle cell migration from the tunica intima to media on days 3 and 7 in groups E and F.

CONCLUSION:

The present study revealed that imatinib mesylate, which was initiated as a prophylactic, systemic pretreatment and continued for seven days, gradually decreased intimal hyperplasia at the anastomosis site.     

伊马替尼治疗Ph阳性慢性粒细胞白血病引起的骨髓形态学变化

目的　观察伊马替尼治疗Ph阳性慢性粒细胞白血病 (CML)引起的骨髓形态学变化 ,并探讨其与血液学、遗传学疗效之间的关系。方法　 117例Ph阳性CML患者 ,包括干扰素治疗失败的慢性期 5 4例、加速期 4 1例、急髓变期 2 2例 ,日服伊马替尼 4 0 0或 6 0 0mg,持续 18个月以上。结果　治疗18个月内 ,各期患者骨髓增生程度显著降低、原始粒细胞 早幼粒细胞显著减少、骨髓无CML特征的比例显著增加 ,慢性期和加速期患者粒、红细胞比例显著降低、巨核细胞数量显著减少 (P值均 <0 0 5 )。获得血液学疗效者骨髓形态持续正常。治疗中发生骨髓增生低下或极度低下还是增生活跃以上与血液学和遗传学疗效密切相关 :慢性期患者其遗传学有效率分别为 5 8.8%和 86 .5 % (P =0 .0 35 ) ,加速期患者血液学完全缓解率分别为 2 6 .3%和 75 .0 % (P =0 .0 0 4 ) ,急变期患者生存期短于 6个月者比例分别为 77.8%和 16 .7% (P =0 .0 0 9)。在CML进展期 ,治疗 1个月时骨髓形态学无CML特征与有CML特征者相比 ,加速期患者 18个月疾病进展率显著降低 (分别为 2 5 .0 %和 75 .0 % ,P =0 .0 2 8)、总生存率显著升高 (分别为 75 .0 %和 11.8% ,P =0 .0 0 4 ) ;急变期患者获得血液学效应的比例显著增加 (分别为 10 0 .0 %和 4 0 .0 % ,P =0     

Optimal Time Points for BCR-ABL1 Tyrosine Kinase Domain Mutation Analysis on the Basis of European LeukemiaNet Recommendations in Chronic Myeloid Leukemia

BackgroundIn this study we aimed to evaluate appropriate time points for mutation analysis of chronic myeloid leukemia.Patients and MethodsIn total, 961 blood samples obtained from 605 chronic-phase chronic myeloid leukemia patients treated with imatinib were subjected to Sanger sequencing to detect BCR-ABL1 mutations. Mutation frequencies at landmark time points (3, 6, and 12 months) were assessed with 16 landmark responses defined by European LeukemiaNet and 2 additional responses, including a complete hematologic response (CHR) at 3 months and a complete cytogenetic response (CCyR) at 12 months.ResultsAfter 12 months of imatinib treatment of 605 patients, 28 (4.6%) patients harbored 33 mutations, including 23 (69.7%) highly resistant T315I and P-loop mutations. Sequencing data from 650 samples were compared with cytogenetic responses. The mutation frequencies in optimal, warning, and failure groups were 0.5% (2/430), 1.8% (2/110), and 19.1% (21/110), respectively. The molecular response was assessed using 956 samples, and the mutation frequencies were 0.7% (3/425), 3.4% (12/358), and 7.6% (14/173) for the optimal, warning, and failure groups, respectively. For the 2 additional responses, the mutation frequencies in patients with CHR at 3 months and with CCyR at 12 months were 0% (0/160) and 1.7% (4/242), respectively. Overall, mutations were frequently detected at 3-month cytogenetic failure (25.0%), 12-month cytogenetic failure (23.2%), and 6-month cytogenetic failure (10.5%) using response–mutation association analysis.ConclusionIrrespective of mutation frequency, failure of achievement of a cytogenetic response should be conducted with appropriate mutation analysis.     

The Outcomes of Chronic Myeloid Leukemia Patients With Molecular Warning Responses During Imatinib Treatment According to the European LeukemiaNet 2013 Recommendations

BackgroundIn the European LeukemiaNet (ELN) 2013 recommendations, chronic myeloid leukemia (CML) patients with warning response (WR) were suggested to be monitored closely continuing with the same tyrosine kinase inhibitor (TKI). Differently, the guidelines of the National Comprehensive Cancer Network considers switching to another TKI as an option.Patients and MethodsWe retrospectively evaluated 73 CML patients receiving first-line imatinib, who were followed and managed in accordance with ELN recommendations. We compared patients with molecular WR with patients with optimal response (OR) and failure regarding short- and long-term outcomes.ResultsThe cumulative major molecular response (MMR) rates in patients with OR were significantly higher at any time point than those achieved by the WR group. Patients with WR at 3 months had significantly inferior failure-free survival (FFS) than optimal responders, but overall survival (OS) was similar. For 6 and 12 months, the WR and OR groups had similar FFS and OS. Twenty of 23 patients with WR at 12 months achieved MMR during imatinib treatment.ConclusionIt takes longer to get to ELN time points with imatinib than second-generation TKIs (2GTKIs). Treatment might fail in a small proportion of the patients with WR during imatinib treatment, but close and careful monitoring and timely switching to 2GTKIs might translate into favorable outcomes. Avoiding early switch to 2GTKIs would prevent patients from experiencing potential toxicities. There is still a need for prospective comparative studies (ie, continuing imatinib treatment vs. switching to 2GTKIs) in patients with WR, to justify the validity of this response category and to explore the benefit of treatment change in these patients.     

Tratamiento de la enfermedad de injerto contra huésped crónica esclerodermiforme con imatinib: una perspectiva dermatológica

Introduction

Chronic graft-versus-host disease (cGVHD) is the most important cause of late non-relapse mortality after allogeneic hematopoietic stem cell transplantation. Sclerodermatous cGVHD is usually steroid refractory and remains a therapeutic challenge. Activating antibodies against the PDGFR have been reported in patients with sclerodermatous cGVHD. These antibodies induce PDGFR phosphorylation and lead to fibrosis. There is increasing evidence of successful treatment of sclerodermatous cGVHD with imatinib, a tyrosine kinase inhibitor.

伊马替尼对BCR/ABL融合基因阳性的急性巨核细胞白血病原代细胞凋亡作用的研究

目的:研究酪氨酸激酶抑制剂伊马替尼(imatinib)对BCR/ABL融合基因阳性的急性巨核细胞白血病原代细胞凋亡的作用,探讨急性巨核细胞白血病的治疗。方法:报告1例急性巨核细胞白血病的临床及实验室特征;应用骨髓细胞直接法和24h短期培养法制备染色体和R显带技术进行细胞遗传学分析;多重RTPCR方法同时检测29种白血病融合基因。取患者骨髓白血病细胞,台盼蓝拒染实验观察伊马替尼对细胞生长曲线的影响;流式细胞仪分析细胞周期。结果:①该例患者的染色体核型为46,XX,t(1;16)(q22;q22),未见Ph染色体;表达BCR/ABLb2a2mRNA,临床治疗效果差,生存期短。②伊马替尼可以抑制该原代白血病细胞的生长,促其凋亡,使大部分细胞受阻于G0/G1期。结论:伊马替尼可以诱导BCR/ABL阳性急性巨核细胞白血病原代细胞的凋亡。     

直肠间质瘤伊马替尼新辅助治疗后经肛门内镜显微手术切除：附13例报告

目的:评价伊马替尼新辅助治疗结合经肛门内镜显微手术(TEM)局部切除治疗直肠间质瘤(GIST)的疗效和安全性。方法:入组13例直肠GIST患者,先予以伊马替尼治疗30~90 d,肿瘤退缩后予以TEM局部切除肿瘤,分析患者术前临床病理资料、新辅助治疗反应性及相关手术指标。结果:接受伊马替尼新辅助治疗3个月内所有患者均出现缓解,肿瘤降期显著。所有患者除1例改行腔镜下经腹直肠局部切除外,均行TEM局部切除,其中9例的患者(70%)在接受伊马替尼治疗2个月内进行手术。平均手术时间45 min,平均术后住院时间6.7 d。11例患者(85%)手术出血少于20 m L,无术后严重出血或感染病例。无治疗相关死亡或其他严重不良事件。12例TEM患者随期无复发或死亡。结论:部分局部进展期直肠GIST,伊马替尼新辅助治疗后结合TEM能够获得理想的手术效果,安全性高,手术创伤小,是一种可选择的治疗方式。