Myelodysplastic Syndrome with Ph Negative Monosomy 7 Chromosome following Transient Bone Marrow Dysplasia during Imatinib Treatment for Chronic Myeloid Leukemia

Abstract Background VASP (vasodilator-stimulated phosphoprotein) and Zyxin are actin regulatory proteins that control cell-cell adhesion. Zyxin directs actin assembly by interacting and recruiting VASP to specific sites of adhesion. The phosphorylation of VASP modifies their activity in cell-cell junctions. PKA phosphorylates VASP at serine 157 regulating VASP cellular functions. VASP is a substrate of BCR-ABL oncoprotein and is tyrosine-phosphorylated in leukemic cells. However, the function of VASP and Zyxin in hematopoietic cells, in the BCR-ABL pathway and its participation in chronic myeloid leukemia (CML) remains unknown. Aims To analyze VASP and Zyxin expression in bone marrow cells from CML patients and healthy donors, as well the involvement of these proteins in hematopoietic cell differentiation and in the BCR-ABL signaling pathway. Materials and Methods VASP and Zyxin expression and phosphorylation were studied in bone marrow samples from 29 individuals (5 healthy donors, 5 CML patients at diagnosis, 16 CML patients responsive to treatment with tyrosine kinase inhibitors (ITK) and 3 CML patients resistant to ITK). One patient was analyzed at diagnosis and after ITK response. VASP or Zyxin silencing was performed by shRNA-lentiviral delivery in K562 cell line, an appropriated shControl was used. ShControl, shVASP and shZyxin K562 cells were induced to megakaryocytic differentiation with 20nM of PMA (phorbol myristate -13 -12 acetate) during 4 days and CD61 expression, a marker for maturing megakaryocytes, was verified by flow cytometry. During megakaryocytic differentiation, VASP and Zyxin gene expressions were evaluated by quantitative PCR; protein expression and activation were determined by Western blotting. Effector proteins of proliferation, apoptosis and adhesion in the BCR-ABL signaling pathway were analyzed in cells silenced for VASP or Zyxin. The interaction of VASP and BCR-ABL or FAK was evaluated by co-immunoprecipiation. Results Healthy donors showed p-VASP ser157 expression, in contrast to CML patients at diagnosis who did not present phospho-VASP ser157. After Imatinib treatment CML patients restored VASP phosphorylation however resistant patients maintained this absence. Zyxin showed the same expression in patients and healthy donors. During Imatinib treatment of K562 cells, phospho-VASP ser157 expression was increased and its interaction with BCR-ABL protein was reduced. VASP and Zyxin gene expressions were upregulated during megakaryocyte differentiation of K562 cells (8.7-fold increase, P=0.0115, and 3.6-fold increase, P=0.015, respectively). VASP and Zyxin protein expressions were increased during megakaryocytic differentiation, including the active form of these proteins (p-VASP ser157 and p-Zyxin ser142). VASP silencing in K562 cells resulted in a 40% decrease of CD61 expression at the end of the megakaryocytic differentiation (P<0.05). In addition, VASP and Zyxin silencing resulted in a decrease of BCL-2 and BCL-XL proteins. VASP binds to FAK, an adhesion effector protein of the BCR-ABL pathway, and it´s silencing resulted in a decreased phosphorylation of FAK y925. Conclusions In BCR-ABL cells, VASP and Zyxin modulated anti-apoptotic proteins and megakaryocytic differentiation. Hence, the altered expression of VASP activity in CML patients may contribute to the pathogenesis of the disease, affecting cellular differentiation or leukemic cell adhesion. Disclosures: No relevant conflicts of interest to declare.

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Acute Myeloid Leukemia with Basophilic Differentiation Transformed from Myelodysplastic Syndrome

Case Reports in Hematology ◽

10.1155/2017/4695491 ◽

2017 ◽

Vol 2017 ◽

pp. 1-6 ◽

Cited By ~ 1

Author(s):

Yasuhiro Tanaka ◽

Atsushi Tanaka ◽

Akiko Hashimoto ◽

Kumiko Hayashi ◽

Isaku Shinzato

Keyword(s):

Bone Marrow ◽

Acute Myeloid Leukemia ◽

Myelodysplastic Syndrome ◽

Myeloid Leukemia ◽

Bone Marrow Failure ◽

Immunosuppressive Treatment ◽

Flow Cytometric Analysis ◽

Chromosomal Analysis ◽

Monosomy 7 ◽

Acute Myeloid

Myelodysplastic syndrome (MDS) terminally transforms to acute myeloid leukemia (AML) or bone marrow failure syndrome, but acute myeloid leukemia with basophilic differentiation has been rarely reported. An 81-year-old man was referred to our department for further examination of intermittent fever and normocytic anemia during immunosuppressive treatment. Chromosomal analysis showed additional abnormalities involving chromosome 7. He was diagnosed as having MDS. At the time of diagnosis, basophils had not proliferated in the bone marrow. However, his anemia and thrombocytopenia rapidly worsened with the appearance of peripheral basophilia three months later. He was diagnosed as having AML with basophilic differentiation transformed from MDS. At that time, monosomy 7 was detected by chromosomal analysis. We found that basophils can be confirmed on the basis of the positivity for CD203c and CD294 by flow cytometric analysis. We also found by cytogenetic analysis that basophils were derived from myeloblasts. He refused any chemotherapy and became transfusion-dependent. He died nine months after the transformation. We should keep in mind that MDS could transform to AML with basophilic differentiation when peripheral basophilia in addition to myeloblasts develops in patients with MDS.

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Evaluation of Ph+/CD34+ Residual Cells in Chronic Myeloid Leukemia Patients after Long Lasting Treatment with Imatinib Mesylate.

Blood ◽

10.1182/blood.v106.11.1999.1999 ◽

2005 ◽

Vol 106 (11) ◽

pp. 1999-1999

Author(s):

Monica Bocchia ◽

Elisabetta Abruzzese ◽

Micaela Ippoliti ◽

Simona Calabrese ◽

Alessandro Gozzetti ◽

...

Keyword(s):

Bone Marrow ◽

Chronic Myeloid Leukemia ◽

Imatinib Mesylate ◽

Median Time ◽

False Positive ◽

Myeloid Leukemia ◽

Prolonged Treatment ◽

Published Data ◽

Imatinib Treatment ◽

Cd34 Cells

Abstract Although the success of imatinib mesylate therapy represents an exciting advance in targeted cancer therapy, it has still to be determined whether responses to this p210 inhibitor in chronic myeloid leukemia (CML) patients will be durable. In fact most of clinical studies agree on the evidence of a persistent molecular disease in the majority of treated patients and altough the absolute level of bcr-abl transcript may vary over the treatment, yet a molecular complete response is of rare observation. In addition, discontinuation of imatinib exerts always in rapid loss of response. In accordance to this the persistence of malignant progenitors in patients in complete cytogenetic response (CCR) after short term imatinib treatment, has been recently demonstrated. In particular, Bathia et al. showed in 12/15 patients studied after a median time of 10 months of imatinib treatment a median of 11% of residual CML CD34+ progenitors in the bone marrow (by FISH Dual Fusion bcr/abl analysis)while only 3/15 patients had no detectable residual CD34+ cells. Less is known about residual Ph+/CD34+ cells surviving after a prolonged therapy with this targeting drug. Thus, we evaluated the amount of bone marrow residual CD34+ cells in 17 CML patients in stable CCR after a long lasting treatment with imatinib. At the time of evaluation, the patients were on conventional dose (400mg) Imatinib for a median time of 48 months (range 36–58 months) having achieved a CCR status (conventionally defined as the complete absence of t(9;22) on caryotypic analysis) within 3 to 6 months of treatment. However all of them still showed molecular disease as detected by nested RT-PCR. Bone marrow CD34+ cell-enriched populations were selected from mononuclear cells using immunomagnetic column separation and were evaluated after cytospin by FISH using a bcr-abl Dual Color Extra Signal Probe(LSI bcr-abl ES, Vysis), that is able to detect bcr-abl fusion in interphase nuclei with a false positive signal rate close to 0. A minimum of 100 CD34+ nuclei per each sample were evaluated. Interestingly, in 8/17 patients no Ph+/CD34+ cells were detected, while in the remaining 9/17 patients a median of 2% (range 0.5–11%) of bcr-abl positive progenitors were still observed. In this small selected serie of patients prolonged treatment with imatinib appears to be correlated with a lower, yet detectable, amount of residual bone marrow Ph+/CD34+ cells when compared to previously published data. This result could be partly explained with the different specificity and sensitivity of the probe used (bcr/abl ES<1% false positive; bcr-abl Dual Fusion 8–10% false positive) The clinical significance of these data as well as the role of this cell target to monitor minimal residual disease in CML needs to be evaluated on a larger serie of patients.

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The Importance of Bone Marrow Lymphocyte Subtypes As Predicting Factors for Molecular Recurrence in Patients with Chronic Myeloid Leukemia after Discontinuation of Imatinib

Blood ◽

10.1182/blood-2021-149242 ◽

2021 ◽

Vol 138 (Supplement 1) ◽

pp. 2564-2564

Author(s):

Arthur Gomes Oliveira Braga ◽

Katia B Pagnano ◽

Marina Dal'Bó Pelegrini Campioni ◽

Ana Beatriz P Lopez ◽

Konradin Metze ◽

...

Keyword(s):

Bone Marrow ◽

Chronic Myeloid Leukemia ◽

Board Of Directors ◽

Median Time ◽

Myeloid Leukemia ◽

Lymphocyte Subsets ◽

Chronic Phase ◽

Imatinib Treatment ◽

Advisory Committees ◽

Treatment Free Remission

Abstract Introduction: in recent years the feasibility of the discontinuation of tyrosine kinase treatment in chronic myeloid leukemia (CML) has been proven, and several clinical factors influencing the duration of treatment-free remission (TFR) after discontinuation have been studied. Aim: we analyzed the influence of bone marrow (BM) lymphocyte subsets on the molecular recurrence after discontinuation of Imatinib (IM) in CML. Methods: in a recent discontinuation study performed at our Institution (EDI-PIO trial) we assessed BM lymphocyte subsets before and after introduction of pioglitazone which was given 3 months before discontinuation of IM. Criterias for discontinuation were: patient in chronic phase at diagnosis, a minimum of 3 years on TKI and sustained molecular remission MR4.5 for at least 2 years. Lymphocyte populations studied: B, TCD8, TCD4, T CD4+CD8+ and T CD4- CD8- besides T naïve and memory, TCRαβ, TCRγδ, NK-t and NK cells. The influence of Sokal score at diagnosis, the duration of imatinib treatment together with the BM lymphoid subsets on the time of treatment-free remission (TFR) were examined by uni- and multivariate Cox regressions. Results: we studied 30 out of 32 patients diagnosed between 1998 and 2013 that reached criteria for discontinuation that were included in EDI-PIO trial: 13 male and 17 female. Median age at diagnosis: 41 years (22-65). Median time of IM treatment: 116.3 months (38.2-209.3); 11 patients (36%) had a molecular recurrence in a median of 5,17 months (2.4-29.4). For patients remaining in TFR the median time of follow-up was 46 months (26.3-55.9). Median overall time of IM treatment (IM-Tr) was 65 months for patients recurring and 124 months for those remaining in TFR. In the univariate Cox regression a significant value was found for IM-Tr, higher percentages of T CD4+CD8+ and lower ones of TCRγδ lymphocytes at discontinuation. In the multivariate model only the T CD4+CD8+ remained. Conclusion: discontinuation of IM is feasible in patients remaining in continuous MR4.5 for more than 2 years. A longer time of IM treatment and higher values of BM T CD4+CD8+ predict a lower risk of relapse. Key words: chronic myeloid leukemia, imatinib, discontinuation, lymphocytes, pioglitazone Disclosures Pagnano: Novartis: Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Astellas: Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Pintpharma: Other: Lecture; EMS: Other: Lecture; Jansenn: Other: Lecture.

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Myelodysplasia and Acute Myeloid Leukemia during Treatment of Chronic Myeloid Leukemia with Dasatinib

Blood ◽

10.1182/blood.v112.11.4284.4284 ◽

2008 ◽

Vol 112 (11) ◽

pp. 4284-4284

Author(s):

Monika Conchon ◽

Israel Bendit ◽

Patricia Ferreira ◽

Lucia Dias ◽

Cristina Kumeda ◽

...

Keyword(s):

Bone Marrow ◽

Myeloid Leukemia ◽

Kinase Inhibitors ◽

Chemotherapeutic Agents ◽

Imatinib Treatment ◽

Previous Exposure ◽

Positive Clone ◽

Monosomy 7 ◽

Retrospective Assessment ◽

Acute Myeloid

Abstract The incidence and significance of Philadelphia negative aberrant clones in patients treated with the second generation of tyrosine kinase inhibitors, including dasatinib, are infrequent. Here we report 2 patients with myelodysplastic syndrome associated with monosomy 7 in Philadelphia negative cells during consecutive imatininb and dasatinb therapy. No previous exposure to chemotherapeutic agents was reported in both patients. In case 1, retrospective assessment of the bone marrow sample collected prior to treatment with dasatinib did not show any abnormal clone, which became evident only after dasatinib was commenced. In case 2, monsomy 7 were found already in 15% Philadelphia negative cells during imatinib treatment and persisted during dasatinib therapy. In the 2 cases described, we assumed that the consecutive imatinib/dasatinib therapy, could abrogate the proliferation of the Philadelphia positive clone and allow pre-existing monosomy 7 clone to emerge.

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Late Development of Cytogenetic Abnormalities in Ph Negative Cells of Chronic Myeloid Leukemia Patients Treated with Imatinib.

Blood ◽

10.1182/blood.v114.22.1108.1108 ◽

2009 ◽

Vol 114 (22) ◽

pp. 1108-1108

Author(s):

Michael Deininger ◽

François-Xavier Mahon ◽

Francois Guilhot ◽

Giuseppe Saglio ◽

Philipp le Coutre ◽

...

Keyword(s):

Chronic Myeloid Leukemia ◽

Acute Leukemia ◽

Cytogenetic Analysis ◽

Myeloid Leukemia ◽

Conflicts Of Interest ◽

Imatinib Treatment ◽

Trisomy 8 ◽

Monosomy 7 ◽

Late Event

Abstract Abstract 1108 Poster Board I-130 Clonal cytogenetics abnormalities in Ph negative metaphases (CCA/Ph-) are observed in a variable fraction of Chronic Myeloid Leukemia (CML) patients (pts) after they obtain a Complete Cytogenetic Remission (CCyR). It is not known whether such abnormalities develop soon after CCyR or if they can appear as a late event. Cytogenetic analysis remains the only methodology able to detect such abnormalities and its use in pts in CCyR after several years of imatinib therapy is being questioned. The Imatinib Long Term Effect (ILTE) study enrolled 948 CML pts in 24 centers around the world (Europe, North/South America, Africa, Middle East and Asia); in order to be eligible, pts had to achieve a CCyR within 2 years after starting imatinib. These pts are being followed for long term side effects such as loss of CCyR, toxicities including second cancers, and survival. Within the ILTE cohort, 384 eligible pts received imatinib for > 5 years and remained in CCyR at 5 years. In 309 cases, at least one standard routine cytogenetic analysis after 5 years of treatment was available. The median duration of imatinib treatment is 6.5 years in this group of pts. A cytogenetic abnormality in the Ph negative metaphases was detected in 18 cases (5.8%; 99% Confidence Interval: 0-10.1%); the number of available cytogenetics analyses positive for CCA/Ph- varied from 1 to 12 per patient. The percentages of pts positive for CCA/Ph- in the the different participating centers ranged between 0 and 28.6%. Of the 18 cases positive for CCA/Ph-, 10 were diagnosed within the first 5 years of treatment, and 8 cases afterwards. Three pts (17%) developed abnormalities during the first 2 years of treatment, 5 (28%) during the third or fourth year, 4 (22%) during the fifth or sixth year, and 6 (33%) during year 7, 8 or 9. Abnormalities were: deletion of Y chromosome (7 cases), trisomy 8, del 7q (2 cases each), monosomy 7, trisomy 6, del 9q, Y duplication, del 13, del 18. With a median follow up of 4.5 years after first detection, none of the patients have developed acute leukemia or myelodysplasia. In addition none of these 18 pts lost his/her CCyR status. CCA/Ph-are detectable in a low but consistent proportion of CML pts in CCyR; their occurrence is not limited to the first 5 years of treatment. Our study supports the notion that patients with CCA/Ph- have a favorable prognosis, despite the similarity of the abnormalities to those observed in acute leukemia and myelodysplasia, suggesting CCA/Ph- to be quite slow in their evolution. These data suggest that the search for CCA/Ph- should not be limited to the first years of imatinib treatment. Disclosures No relevant conflicts of interest to declare.

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