Focal lymphoblastic transformation of chronic myelogenous leukemia develops into erythroid leukemia:A case report

BACKGROUND We present a case of focal lymphoblastic transformation to erythroid leukemia following acute myeloblastic transformation in a patient with chronic myelogenous leukemia(CML)and discuss its mechanism of occurrence and development.CASE SUMMARY The presence of the Philadelphia(Ph)chromosome was identified through karyotype analysis,while the BCR-ABL fusion gene was detected using quantitative real-time polymerase chain reaction of the peripheral blood sample.Fluorescence in situ hybridization was used to detect the expression of the BCRABL gene in the lymphoma.Antigen expression and gene mutations in the primitive cells were detected by flow cytometry.The analysis confirmed the presence of CML along with focal lymphoblastic transformation to erythroid leukemia.Additionally,the patient was found to have secondary erythroid leukemia,along with multiple new gene mutations and abnormalities in complex karyotypes of chromosomes.CONCLUSION Our findings suggest a possible molecular basis for the focal lymphoblastic transformation secondary to myeloblastic transformation in patients with CML.

Overexpression of P-glycoprotein induces acquired resistance to imatinib in chronic myelogenous leukemia cells

Imatinib,a breakpoint cluster region(BCR)-Abelson murine leukemia(ABL) tyrosine kinase inhibitor(TKI),has revolutionized the treatment of chronic myelogenous leukemia(CML).However,development of multidrug resistance(MDR) limits the use of imatinib.In the present study,we aimed to investigate the mechanisms of cellular resistance to imatinib in CML.Therefore,we established an imatinib-resistant human CML cell line(K562-imatinib) through a stepwise selection process.While characterizing the phenotype of these cells,we found that K562-imatinib cells were 124.6-fold more resistant to imatinib than parental K562 cells.In addition,these cells were cross-resistant to second-and third-generation BCR-ABL TKIs.Western blot analysis and reverse transcription-polymerase chain reaction(RT-PCR) demonstrated that P-glycoprotein(P-gp) and MDR1 mRNA levels were increased in K562-imatinib cells.In addition,accumulation of [14C]6-mercaptopurine(6-MP) was decreased,whereas the ATP-dependent efflux of [14C]6-MP and [3H]methotrexate transport were increased in K562-imatinib cells.These data suggest that the overexpression of P-gp may play a crucial role in acquired resistance to imatinib in CML K562-imatinib cells.

Anti-proliferative effects of a small molecule inhibitor of CDKAT7519 on chronic myeloid leukemia (CML) cells through haltingthe transition of cells from G2/M phase of the cell cycle

Pathogenesis of chronic myeloid leukemia(CML)has mostly been studied with regard to the oncogenic role of BCR/ABL fusion,however,recent disclosures have declared that the challenges with the treatment of CML patients would not be resolved until the role of other aberrancies is ignored.Given the involvement of cyclin-dependent kinases(CDKs)in the pathogenesis of CML,the present study aimed to investigate the effects of a multi-CDK inhibitor AT7519 on BCR/ABL-harboring CML-derived K562 cells.Our results showed that AT7519 effectively reduced the survival of K562 and induced its anti-proliferative effect through the induction of G2/M arrest due to elevated p21 and p27.The resulting data also revealed that either direct or indirect suppression of c-Myc using specific c-Myc inhibitor 10058-F4 and selective PI3K inhibitor CAL-101 resulted in a superior cytotoxicity,suggesting that the activation of PI3K pathway could attenuate antileukemic effects of the inhibitor,at least partly,through a c-Mycdependent mechanism.To the best of our knowledge,to date,no study has addressed the effect of autophagy on CML cell response to AT7519,and,herein,we proposed for the first time that the suppression of autophagy boosted AT7519 cytotoxicity against K562.Overall,we suggested that selective CDK inhibitor AT7519 exerted antileukemic effect against CML cells and propose a novel therapeutic application for the inhibitor either as a single agent or in combination with c-Myc and/or PI3K inhibitors.

Myeloid sarcoma presenting as a colon polyp and harbinger of chronic myelogenous leukemia

Myeloid sarcoma, also known as granulocytic sarcoma or chloroma is an unusual accumulation of malignant myeloid precursor cells in an extramedullary site, which disrupts the normal architecture of the involved tissue. It is known to occur more commonly in patients with acute myelogenous leukemia and less commonly in those with myelodysplastic syndrome and myeloproliferative neoplasm, such as chronic myelogenous leukemia. The most common sites of involvement include bone, skin and lymph nodes. However, rare cases have been reported in the gastrointestinal tract, genitourinary tract, or breast. Most commonly, a neoplastic extramedullary proliferation of myeloid precursors in a patient would have systemic involvement of a myeloid neoplasm, including in the bone marrow and peripheral blood. Infrequently, extramedullary disease may be the only site of involvement. It may also occur as a localized antecedent to more generalized disease or as a site of recurrence. Herein, we present the first case in the English literature of a patient presenting with an isolated site of myeloid sarcoma arising in the form of a colonic polyp which, after subsequent bone marrow biopsy, was found to be a harbinger of chronic myelogenous leukemia.

Redistribution of Platelet Membrane Glycoprotein IV and Release of Intracellular α-granule Thrombospondin in Patients with Chronic Myelogenous Leukemia

The redistribution of platelet membrane glycoprotein IV (GPIV) and the release of intracellular Q-granule thrombospondin (TSP) were examined and the inhibition of 5-thromboglobulin (&TG) and platelet factor 4 (PF4) in patients with chronic myelogenous leukemia (CML) was observed and quantitation of β-TG and PF4 in sera was conducted. GPIV in inactive platelet from CML was 36080±17010 molecules/platelet as compared with 13190±4810 from the controls (P<0,01), No abnormality was found in the distribution of platelet membrane GPIb and GPIIb/III.(P>0. 05). The GPIV redistribution on active platelet membrane induced thrombin (1U/ml) from CML and healthy donors was 44320132310 and 228001 12700 molecules/platelet respectively (P<0. 01 ). The difference in the release of intracellular Q-granule TSP between CML and the control group was not found (P>0.05). There was no direct correlation between GPIV expression and TSP binding after platelet activation. The high leveIs of β-TG and PF4 in sera inhibited release of intracellular a-granule TSP in vitro. These results indicate that the abnormality of platelet membrane GPIV is a common marker in CML, therefore the specific increase of platelet GPIV in patients with CML may be a useful tool for the diagnosis and monitoring of the platelet dysfunction. The release of interna1 TSP pools is hindered by either β-TG or PF4 in sera.

Preliminary Research on the p53 Gene Rearrangements in the Evolution of Chronic Myelogenous Leukemia to Blast Crisis

DNA from 36 patients with chronic myelogenous leukemia (CML) at various clinical stages and 6 cases of acute leukemia was investigated for alterations of the p53 gene by Southern blot analysis.Rearrangements of the p53 gene were seen in 3 of 12 (25.00%) cases of blast crisis and accelerated phase (AP) of CML and in only one of 18 chronic phrase (CP),just as has been reported previously. Meanwhile,by restriction fragment length polymorphism (RFLP) analysis the Bgl II site polymorphism in the p53 gene was also found. The frequency in Chinese people detected here was 0.392,which was strikingly higher than that in some other countries(P<0. 001).These results suggested that the alterations of the p53 gene, for example,p53 rearrangements,were probably responsible for the progression of BC in some CML patients, and that the frequency of Bgl II polymorphism in the p53 gene might be related to the population distribution.

HIM_(1) AND HIM4, TWO MONOCLONAL ANTIBODIES POTENTIALLY USEFUL FOR AUTOLOGOUS BONE MARROW TRANSPLANTATION IN CHRONIC MYELOGENOUS LEUKEMIA

We developed two complement-fixing MoAbsHIMand HIM(murine)that were specifically reac-tive with chronic myelogenous leukemia (CML) cells.They were capable of fixing human or rabbit com-plement and suitable for CML cells purging of re-mission marrow from CML patients.HIMreactedwith majority leukemic cells form 7 out of 10 CMLpatients by complement-mediated cytotoxicity(C’MC)assay(positive cells 80%—90%),HIMreacted withmajority CML cells from 4 out of 5 CML by C’MCassay(positive cells 80%—90%).Treatment withHIMor HIMand human C’was capable of lysing97% of K562,U937,HL-60 and CML cells in a 20fold excess of unrelated cells by indirect FITC+EBstain.Using limited dilution culture,incubation withHIMand C’produced 1.5 logs inhibition of growthin K562 cells,and 1.9 logs in U937 cells,and withHIMand C’produced 2.9 logs inhibition in HL-60cells and 3.0 logs in U937 cells.Both MoAbs cocktailwas shown 1.8 logs in K562 cells and 3.2 logs in U937cells.They were no suppression on the growth o

Research on Marketing Strategies of Product D—a Chronic Myelogenous Leukemia Drug for Pharmaceutical Company A

Objective To put forward some suggestions on the marketing strategies for chronic myelogenous leukemia in a pharmaceutical enterprise.Methods Based on the development status of the pharmaceutical industry and the SWOT analysis of a company’s product,the marketing strategies were formulated to provide theoretical basis for pharmaceutical enterprise to adapt to the new medical reform.Results and Conclusion Nowadays,due to fierce competition,in order to expand the new market,enterprises should implement the strategies of new products,centralized management and professional training.Meanwhile,the effective marketing strategies should be formulated and strictly carried out according to the conditions of the pharmaceutical company.

Cryptococcal Meningitis in Patient with Chronic Myeloid Leukemia

Objective: This study aimed to report the case of a female patient with chronic myeloid leukemia affected by cryptococcal meningitis. Case report: ML, white, 48 years old, female sex, previously diagnosed with chronic myeloid leukemia that has been refractive to the use of imatinib and who has recently begun using nilotinib, was admitted complaining of sudden and disabling migraine in the last 1 month associated with asthenia, adinamia, anorexia, disinterest for daily activities, dizziness, nausea, and vomiting. She evolved with ataxia, and started to stroll with help and showed decrease of muscular strength in her upper limbs. She also presented episodes of decrease of consciousness, with look fixation, no respond to sound stimulation, and short-term hearing loss. The cerebrospinal fluid showed presence of Cryptococcus sp. and, therefore, we began treatment with intravenous liposomal amphotericin B in the dose of 3 mg/kg/day, for 6 weeks. A new cerebrospinal fluid analysis, at the end of treatment, also showed rare structures that are compatible with Cryptococcus sp. As sequelae, she continued with hearing loss in her right ear and enhancement in her right auditory canal, seen in the magnetic resonance imaging. After stabilization and clinical improvement, she was discharged. After 3 weeks, she was hospitalized again with degeneration of the condition, and died due to intracranial hypertension secondary to cryptococcal infection. Final Considerations: This report reinforces the need of reflecting on fungi pathologies, especially in immunosuppressant patients, as well as the importance of early diagnosing and making a fast intervention, with the aims of providing quality of life and comfort to the patient and of minimizing neurological sequelae to the patient.

Predictive indicators of successful tyrosine kinase inhibitor discontinuation in patients with chronic myeloid leukemia

Clinical trials have demonstrated that some patients with chronic myeloid leukemia(CML)treated for several years with tyrosine kinase inhibitors(TKIs)who have maintained a molecular response can successfully discontinue treatment without relapsing.Treatment free remission(TFR)can be reached by approximately 50%of patients who discontinue.Despite having similar levels of deep molecular response and an identical duration of treatment,the factors that influence the successful discontinuation of CML patients remain to be determined.In this review we will explore the factors identified to date that can help predict whether a patient will successfully achieve TFR.We will also discuss the need for the identification of predictive biomarkers associated with a high probability of achieving TFR for the future personalized identification of patients who are suitable for the discontinuation of TKI treatment.

Possible Evolution of Paroxysmal Nocturnal Hemoglobinuria into Chronic Myeloid Leukemia: A Rare Transformation

PNH is a rare acquired clonal hematopoietic stem cell disorder characterized by abnormal sensitivity of red blood cells to lysis by complement. It is caused by genetic mutation resulting in deficiency of glycosyl phosphatidylinositol anchor (GPA) for cell membrane proteins including complement regulating proteins CD55 and CD59. PNH tends to be associated with Aplastic Anemia (anemia due to failure of the bone marrow to produce red and white blood cells as well as platelets), Myelodysplastic Syndrome (a group of cancers in which immature blood cells in the bone marrow do not mature or become healthy blood cells) or rarely Acute Myeloid Leukemia (AML) (also known as acute nonlymphocytic leukemia, representing a group of clonal hematopoietic stem cell disorders in which both a block in differentiation and unchecked proliferation result in the accumulation of myeloblasts at the expense of normal hematopoietic precursors). Here we report a case and assume possible evolution of PNH into CML (a myeloproliferative malignant clonal disease characterized by presence of fusion BCR/ABL fusion oncogene).

A novel cytogenetic abnormality r(7)(::p11.2->q36.3::) in a Philadelphia-positive chronic myeloid leukemia case

The so-calledPhiladelphia(Ph) chromosome is present in more than 90% of chronic myeloid leukemia (CML) cases. It results in juxtaposition of the 5' part of the BCR gene on chromosome 22 and the 3' part of the ABL1 gene on chromosome 9. An additional acquired monosomy 7 or deletion of 7q is associated with poor prognosis in a variety of myeloid disorders. Here we report a novel Ph chromosome positive CML case with a ring chromosome 7 [r(7)]. Immunophenotyping was compatible with CML, although 4.5% of total leucocytes appeared like acute myelogeneous leukemia (AML) subtype M2. The r(7) was characterized in detail by array-proven multicolor banding (aMCB), the latter being of enormous significance to characterize breakpoint regions in detail. Underlying mechanisms and prognostic are discussed, as ring chromosomes are rare cytogenetic abnormalities in hematopoietic malignancies.

伊马替尼治疗小儿CML及对BCR-ABL融合基因转归的影响

目的 分析应用伊马替尼治疗小儿慢性粒细胞白血病(CML)及对BCR-ABL融合基因转归的影响。方法选取CML患儿86例,按随机数字表法分为研究组(n=43)和对照组(n=43),对照组予以第一代伊马替尼,研究组予以第二代伊马替尼。对比两组血液学疗效、BCR-ABL融合基因转归情况、血液学不良反应发生率、非血液学不良反应发生率、2年生存率。结果 研究组总缓解率[88.37%(38/43)]较对照组[65.12%(28/43)]高(P<0.05);研究组BCR-ABL^(IS)≤10%达标率[95.35%(41/43)]、BCR-ABL^(IS)≤0.0032%达标率[51.16%(22/43)]较对照组[67.44%(29/43)]、[20.93%(9/43)]高(P<0.05);研究组血液学不良反应发生率[6.98%(3/43)]与对照组[11.63%(5/43)]对比无显著差异(P>0.05);研究组非血液学不良反应发生率[4.65%(2/43)]与对照组[16.28%(7/43)]对比无显著差异(P>0.05);研究组2年生存率[97.50%(39/40)]与对照组[90.48%(38/42)]对比无显著差异(P>0.05)。结论 小儿CML应用第二代伊马替尼药物治疗,可提高血液学疗效,促进BCR-ABL融合基因转归,具有用药安全性,并可改善预后。

β-谷甾醇通过诱导ROS累积造成氧化应激抑制K562/ADR细胞增殖并促进凋亡和分化

目的:探讨β-谷甾醇抑制耐阿霉素人类髓性白血病细胞K562/ADR增殖、促进细胞凋亡与分化的效果及其可能机制。方法:使用细胞毒性试验计算β-谷甾醇对K562/ADR细胞的半数抑制率(IC50),采用CCK-8法检测细胞增殖率,采用流式细胞术检测细胞凋亡率与线粒体膜电位下降率,使用分光光度法检测细胞上清中SOD活力与MDA含量,使用荧光试剂盒检测细胞ROS强度,使用联苯胺染色检测红细胞分化程度,使用蛋白质印迹(Western blot)检测珠蛋白转录因子1(GATA-1)、β-珠蛋白(β-globin)和核因子e2相关因子2(NF-E2)蛋白表达水平。结果:β-谷甾醇对K562/ADR细胞的半数抑制浓度的IC50为163.64μmol/L。与空白组相比,3个浓度的β-谷甾醇均能抑制细胞增殖、促进细胞凋亡与红细胞分化,降低SOD活力、线粒体膜电位,增加MDA与ROS含量,升高GATA-1、β-globin和NF-E2蛋白表达水平(P<0.01)。结论:β-谷甾醇能抑制K562/ADR细胞增殖,促进细胞凋亡,并且诱导红细胞分化,其机制可能是通过促进ROS积累引起氧化应激失衡。

分化抑制因子家族在慢性髓系白血病中的表达及临床意义

目的:探索分化抑制因子(inhibitor of differentiation,ID)家族在慢性髓系白血病(chronic myeloid leukemia,CML)中的表达和启动子甲基化水平,并分析其临床意义。方法:应用定量PCR及定量甲基化特异性PCR的方法检测2010年1月至2017年12月期间江苏大学附属人民医院就诊的非恶性血液病患者(对照组)和CML患者骨髓单个核细胞中ID2/ID3/ID4表达及ID4启动子甲基化水平,通过分组分析ID家族异常的临床意义。结果:ID2及ID3表达在CML患者中均呈现显著上调(P<0.001,P<0.05),而ID4表达在CML患者中呈现显著下调(P<0.01)。其中,接受者操作特征曲线分析揭示ID2表达可作为CML鉴别的潜在分子标志物(AUC=0.895,P<0.001)。CML患者中ID4启动子高甲基化概率显著高于对照组患者(P=0.001),且ID4启动子甲基化与ID4表达呈现负相关(r=-0.424,P=0.002)。通过分组分析发现ID2高表达较易发生于男性患者中(P=0.040);ID4低表达/高甲基化较易发生于加速/急变期患者(P=0.003,P<0.001)。此外,CML加速/急变期患者ID4表达水平低于慢性期患者(P<0.001),而ID4甲基化水平高于慢性期患者(P<0.001)。通过单因素及多因素Logistic回归分析发现ID4高甲基化是CML患者疾病进展的独立危险因素(P=0.007)。结论:ID家族在CML患者中表达态势不同,其中ID2/ID3表达上调;而ID4表达下调,与ID4启动子高甲基化相关。ID4表达/甲基化与CML疾病进展相关,其中ID4甲基化可能是CML疾病进展的独立危险因素。

积雪草苷抑制伊马替尼耐药的K562细胞的生长和迁移

目的研究积雪草苷对慢性粒细胞白血病(chronic myelogenous leukemia,CML)细胞增殖、迁移和周期停滞的作用,并且探讨了其对GATA-1表达的调节。方法以CML细胞系K562、抗伊马替尼(imatinib,IM)K562细胞(K562r)和人源CML细胞为模型,CCK-8测定不同浓度积雪草苷对3种细胞活力的影响。选择0、25、50、100μmol/L积雪草苷处理K562r细胞,CFSE标记测定细胞增殖,Transwell测定细胞迁移,流式细胞仪检测细胞周期,蛋白质印迹检测cyclin A、CDK2、CDK4、cyclin D1、Ki67、PCNA、VEGF、MMP-9和MMP-14蛋白水平。结果与对照组比较,积雪草苷处理可显著抑制K562r细胞增殖和迁移,并促进其细胞周期阻滞。同时我们发现积雪草苷处理可显著上调K562r细胞中GATA-1水平。结论积雪草苷能够抑制K562r细胞增殖、迁移并诱导其细胞周期阻滞,其机制可能与GATA-1的表达上调相关。

慢性粒细胞性白血病患者TKI依从性与临床结局的关系

目的探讨慢性粒细胞性白血病(CML)患者对酪氨酸激酶抑制剂(TKI)的依从性及与临床结局的关系。方法选取2020年1月至2023年6月安徽医科大学附属宿州医院收治的80例CML患者,采用中文版莫里斯基用药依从性量表(MMAS⁃8)评估患者的TKI依从性。根据评估结果将患者分为高依从性组和低依从性组,采用多因素Cox比例风险模型分析两组患者依从性相关因素,并比较两组TKI更换率、主要分子反应(MMR)响应率、无进展生存期、生活质量评分。结果Cox回归模型多因素分析结果显示,男性、年龄<70岁、高收入及维持前线TKI治疗的患者依从性更高,且患者的临床结局显示出更明显的改善(P<0.05)。高依从性组的TKI更换率显著低于低依从性组,差异有统计学意义(P<0.05)。高依从性组的TKI更换率显著低于低依从性组,差异有统计学意义(P<0.05)。高依从性组的MMR响应率显著高于低依从性组,差异有统计学意义(P<0.05)。Kaplan⁃Meier生存分析结果显示,高依从性组(62.35±12.39)无进展生存期显著高于低依从性组患者(39.04±10.07),差异有统计学意义(P<0.05)。高依从性患者的生活质量评分显著高于低依从性患者,差异有统计学意义(P<0.05)。结论高依从性CML患者的临床疗效显著更好,临床上应加强干预措施以提高患者的TKI依从性,从而改善患者的预后。

高三尖杉酯碱诱导K562和CML细胞凋亡及分化的实验研究

目的 明确高三尖杉酯碱 (HHT)治疗慢性粒细胞性白血病 (CML)的机理。方法 应用细胞生长曲线、克隆形成能力、细胞内血红蛋白含量测定、细胞形态 ,结合DNA凝胶电泳、流式细胞仪等方法 ,观察HHT对K5 6 2细胞株及CML慢性期细胞的作用方式。进一步用RT PCR方法研究bcr abl、c myc、max基因在转录水平的改变。结果 低浓度HHT抑制K5 6 2细胞的克隆形成能力并使K5 6 2细胞内血红蛋白含量增加 ;0 .1μmol/L及以上浓度HHT可诱导K5 6 2细胞及CML细胞凋亡 ;细胞周期分析发现细胞阻滞于G1期。c myc基因在加药 2 4h开始下调 ;bcr abl和max基因的表达未见明显改变。结论 低浓度HHT可抑制K5 6 2细胞增殖并诱导其向红系分化 ;超过一定“阈浓度”HHT可诱导K5 6 2细胞和CML慢性期细胞凋亡 ;HHT可通过抑制c

中文版FACT-Leu量表用于CML患者生活质量评估及影响因素分析

目的探讨白血病治疗功能评估量表(FACT-Leu)在中国慢性髓系白血病(CML)患者中的适用性,了解国内CML患者生活质量的现状及影响因素,为医疗、护理干预提高患者生活质量提供参考。方法选取2017年4-5月就诊于四川大学华西医院门诊的确诊为CML患者120例,采用FACT-Leu中文版和自制的生活质量影响因素问卷进行横断面调查,了解生活质量现状。结果共发放问卷120份,回收有效问卷111份,FACTLeu中文版的内在一致性系数Cronbach'sα为0.817,各维度分别为生理状况0.875,社会/家庭状况0.898,情感状况0.735,功能状况0.880,白血病专用条目0.864。不同性别、文化程度、医疗保险种类、自理能力、依从性(未遵医嘱服药次数)的CML患者,其生活质量差异有统计学意义(P<0.05)。其中,女性、生活无法完全自理、未遵医嘱服药次数较多、离异或丧偶是危险因素,城镇职工医疗保险或商业保险、文化程度较高是保护性因素。结论FACT-Leu量表中文版在中国CML患者的生活质量评价中具有良好的信度和适用性;我国CML患者生活质量与性别、文化程度、婚姻状况、服药依从性、生活自理能力有关,提示医疗护理中,应重视对CML患者服药依从性和自理能力的干预,以提高其生活质量。