Clonal Chromosomal Abnormalities Research Articles

Potential role for second‑generation tyrosine kinase inhibitors in patients with chronic myeloid leukemia harboring additional clonal chromosome abnormalities: A retrospective CML Cooperative Study Group analysis.

Tyrosine kinase inhibitor (TKI) treatment is the standard of care for patients with chronic myeloid leukemia (CML). Even in the imatinib era, the presence of 'clonal chromosomal abnormalities' in the Philadelphia chromosome (CCA/Ph+) at diagnosis reportedly increased the risk of disease progression and predicted shorter survival. However, it remains unclear whether CCA/Ph+ is a poor prognostic marker in the era of new‑generation TKIs. The data of patients with CML in the chronic phase (CP) that were extracted from the CML Cooperative Study Group database were retrospectively analyzed. Of the 328 eligible patients, 33 (10.1%) had CCA/Ph+, including 9 major route and 24 minor route aberrations. The characteristics of patients with and without CCA/Ph+ were similar; however, the proportion of blasts was higher among patients with CCA/Ph+. Notably, the survival rate of patients with CCA/Ph+ was not inferior to that of patients without CCA/Ph+, and there were no differences in responses to TKIs. All 9 patients with major route CCA/Ph+ attained a major molecular response (MMR) with no disease progression, and 8 ultimately achieved a deep molecular response. In particular, the median interval between TKI initiation and achievement of MMR was shorter in patients initially treated with a second‑generation TKI than in those treated with imatinib (5 vs. 10 months). The present retrospective study, thus, revealed favorable treatment outcomes in CML‑CP patients with CCA/Ph+ treated with second‑generation TKIs. The data indicated that administering second‑generation TKIs as first‑line treatments is preferable in CML‑CP patients with CCA/Ph+.

Benign inflammatory myofibroblastic tumour of the nose: a rare case report

Inflammatory myofibroblastic tumor (IMT) of the head and neck region are rare and benign and of unknown etiology. They mimic as malignant lesions both clinically and radiologically. It encompasses a spectrum of myofibroblastic proliferation along with varying amount of inflammatory infiltrate. Recently, the concept of this lesion being reactive has been challenged based on the clinical demonstration of recurrences and metastasis and cytogenetic evidence of acquired clonal chromosomal abnormalities. Here we report a case of a 16 year old male who presented with a swelling on the nasal dorsum for 6 months. MRI and FNAC revealed a spindle cell neoplasm. The swelling was resected completely under general anaesthesia. Histopathology with immunohistochemistry proved it to be a benign inflammatory myofibrobalstic tumour. Complete surgical resection is the best treatment modality for IMT.

Clinical implications of clonal chromosomal abnormalities in Philadelphia negative cells in CML patients after treated with tyrosine kinase inhibitors

Emergence of clonal chromosomal abnormalities in Philadelphia chromosome-negative (CCA/Ph-) cells in chronic myeloid leukemia (CML) patients during the treatment with tyrosine kinase inhibitors (TKIs) is an interesting phenomenon. Although previous studies revealed some potential impact of CCA/Ph- on CML patients’ outcome, clinical significance of CCA/Ph- in CML patients remains to be further elucidated. We retrospectively reviewed the patients with CML evaluated at Genoptix Medical Laboratory in Carlsbad, California from 2005 to 2015. Twenty-four CML patients with CCA/Ph- cells were identified. These include 18 patients with single chromosomal abnormality, 4 patients with double chromosomal abnormalities, and two patients with complex cytogenetic abnormalities. In addition to trisomy 8 and monosomy 7, we identified that 20q- was also a common abnormality in CCA/Ph- cells. Most of the patients with CCA/Ph- cells demonstrated no significant dysplasia or increased blasts with two exceptions: one patient with persistent 7q- exhibiting mild dysmegakaryopoiesis, suggestive of an early evolving myelodysplastic syndrome, and another patient with complex cytogenetic abnormalities who developed acute myeloid leukemia after gained MLL amplification. One patient with complex cytogenetic abnormalities showed optimal response to TKI treatment, no overt dysplasia, and no disease progression during almost 4-years of follow-up. More interestingly, FISH tests could identify more cases with double chromosomal abnormalities and these cases showed suboptimal responses to TKI treatments. Our observation indicates that 20q- was also a common abnormality in CCA/Ph- cells, further FISH tests revealed additional CCA/Ph-, and the majority of CML patients with two or more chromosomal abnormalities in Ph- cells showed inferior response to TKI treatments. The results of our study suggest that CML cases with CCA/Ph- may represent a group of patients with heterogeneous genetic alterations.

Philadelphia chromosome-negative myeloid neoplasms in patients with Philadelphia chromosome-positive chronic myeloid leukemia during tyrosine kinase inhibtor-therapy

目的探讨Ph阳性慢性髓性白血病（Ph+CML）患者在酪氨酸激酶抑制剂（TKI）治疗中出现Ph阴性克隆性染色体异常（CCA/Ph−）后发生Ph−骨髓增生异常综合征或急性髓系白血病（MDS/AML）患者的临床特征。方法回顾性分析2001年5月至2017年12月收治的出现CCA/Ph−的CML患者资料。结果共收集63例在TKI治疗过程中检出CCA/Ph−的Ph+CML患者，其中7例发生Ph−MDS/AML，56例疾病稳定。与疾病稳定组相比，首次检出CCA/Ph−时，Ph− MDS/AML组患者血红蛋白（P=0.007）和PLT（P=0.006）水平更低，外周血原始细胞比例（P<0.001）更高；当发生MDS或AML时，Ph−MDS/AML组患者的CCA/Ph−以“−7”为主（5/7，71.4%），而Ph−疾病稳定组以“+8”为多见（32/56，57.1%）。Ph−MDS/AML组患者总体预后不良，而Ph−疾病稳定组多数CCA/Ph−持续存在或有变化，大部分患者分子学反应良好，疾病稳定。结论CML患者接受TKI治疗中，少数会发生CCA/Ph−，其中大部分病情稳定，极少数会发生Ph−MDS/AML。当出现不明原因血细胞减少或CCA/Ph−为“−7”时，应重视外周血涂片、骨髓细胞遗传学和骨髓细胞形态学检测。

Cytogenetic analysis of 130 renal oncocytomas identify three distinct and mutually exclusive diagnostic classes of chromosome aberrations.

The cytogenetic alterations in renal oncocytoma (RO) are poorly understood. We analyzed 130 consecutive RO for karyotypic alterations. Clonal chromosome abnormalities were identified in 63 (49%) cases, which could be categorized into three classes of mutually exclusive cytogenetic categories. Class 1 (N = 20) RO had diploid karyotypes with characteristic 11q13 rearrangement in balanced translocations with 10 or more different chromosome partners in all cases. We identified recurrent translocation partners at 5q35, 6p21, 9p24, 11p13-14, and 11q23, and confirmed that CCND1 gene rearrangement at 11q13 utilizing fluorescence in situ hybridization (FISH). Class 2 RO (N = 25) exhibited hypodiploid karyotypes with loss of chromosome 1 and/or losses of Y in males and X in females in all cases. The class 3 tumors comprising of 18 cases showed diverse types of abnormalities with the involvement of two or more chromosomes exclusive of abnormalities seen in classes 1 and 2 tumors. Furthermore, karyotypically uninformative cases were subjected to FISH analysis to identify classes 1 and 2 abnormalities. In this group, we found similar frequencies of CCND1 rearrangement, loss of chromosome 1 or Y as with karyotypically abnormal cases. We validated our results against 91 tumors from the Mitelman database. Correlation of clinical data with all the three classes of ROs showed no clear evidence of overall patient survival. Our findings support the hypothesis that RO exhibit three principal cytogenetic categories, which may have different roles in initiation and/or progression. These cytogenetic markers provide a key tool in the diagnostic evaluation of RO.

Prognosis of clonal chromosomal abnormalities in Philadelphia negative metaphases cells in chronic myeloid leukemia with tyrosine kinase inhibitor therapy

目的探讨酪氨酸激酶抑制剂（TKI）治疗期间Ph−细胞间期出现克隆性染色体异常（CCA/Ph−）的特征以及对慢性髓性白血病（CML）患者预后的影响。方法回顾性分析河南省肿瘤医院2007年8月至2017年7月期间接受TKI治疗并出现CCA/Ph−的30例CML患者的临床资料。结果30例CML患者，男性19例（63.3%）。首次出现CCA/Ph−时患者中位年龄44（14~68）岁，TKI中位治疗13（2~94）个月。TKI治疗3个月BCR-ABLIS≤10%者占46.7%（14/30）。60.0%（18/30）的患者首次出现CCA/Ph−的克隆比例≥50%。63.3%（19/30）的患者CCA/Ph−呈一过性出现（仅出现1次），36.7%（11/30）呈反复出现（出现次数≥2）。CCA/Ph−类型：+8为主异常占60.0%，−7/7q−为主异常占13.3%，−Y异常占6.7%。中位随访50个月，76.7%（23/30）的患者维持完全细胞遗传学反应；63.3%（19/30）获主要分子学反应，其中43.3%（13/30）达分子学无法检测（UMRD）。30例患者中位无事件生存（EFS）时间为44个月，2年和5年EFS率分别为（82.1±7.3）%和（52.4±12.8）%，中位总生存（OS）时间为50个月，2年和5年OS率分别为（92.6±5.0）%和（77.2±14.7）%。单因素分析显示：男性、CCA/Ph−出现≥2次、3个月BCR-ABLIS>10%的CML患者2年EFS率分别较女性、CCA/Ph−一过性出现、3个月BCR-ABLIS≤10%者显著降低（P值均<0.05）；CCA/Ph−出现≥2次者2年OS率较CCA/Ph−一过性出现者显著降低（P<0.05）。多因素分析显示：CCA/Ph−出现频率是影响CML患者EFS率的独立危险因素（RR=4.741，95%CI 1.21~18.571，P=0.018）。结论TKI治疗期间出现CCA/Ph−以+8最常见，其次为−7/7q−和−Y；CCA/Ph−克隆比例多≥50%；可呈反复出现或一过性出现，以一过性出现更常见。CCA/Ph−反复出现（≥2次）是影响CML患者EFS和OS的独立危险因素。

Cancer chromosome breakpoints cluster in gene-rich genomic regions.

Cancer cells are characterized by chromosome abnormalities, of which some, in particular balanced rearrangements, are associated with distinct tumor entities and/or with specific gene rearrangements that represent important steps in the carcinogenic process. However, the vast majority of cytogenetically detectable structural aberrations in cancer cells have not been characterized at the nucleotide level; hence, their importance and functional consequences are unknown. By ascertaining the chromosomal breakpoints in 22 344 different clonal structural chromosome abnormalities identified in the karyotypes of 49 626 cases of neoplastic disorders we here show that the distribution of breakpoints is strongly associated (P < 0.0001) with gene content within the affected chromosomal bands. This association also remains highly significant in separate analyses of recurrent and nonrecurrent chromosome abnormalities as well as of specific subtypes of cancer (P < 0.0001 for all comparisons). In contrast, the impact of band length was negligible. The breakpoint distribution is thus not stochastic—gene‐rich regions are preferentially affected. Several genomic features relating to transcription, replication, and chromatin organization have been found to enhance chromosome breakage frequencies; this indicates that gene‐rich regions may be more break‐prone. The salient finding in the present study is that a substantial fraction of all structural chromosome abnormalities, not only those specifically associated with certain tumor types, may affect genes that are pathogenetically important. If this interpretation is correct, then the prevailing view that the great majority of cancer chromosome aberrations is cytogenetic noise can be seriously questioned.

Results of Cytogenetic and Molecular Cytogenetic Studies in Relapce/Refractory of Multiple Myeloma

Background. Multiple myeloma (MM) – a tumor of the lymphoid tissue, which is characterized by low proliferative activity of abnormal cells and a wide range of acquired chromosomal aberrations that play a leading role in predicting the course of the disease. Accumulation of new data in the relapsed/refractory of MM will make it possible to understand the nature of the formation of cloning insensitive to chemotherapy (CT), to predict the effectiveness of therapy, which will further improve the individualization of treatment for each patient. Objective. The aim of the study is to determine the peculiarities of chromosomal aberrations of abnormal clones of bone marrow (BM) cell in relapsed/refractory of MM by standard cytogenetic and molecular cytogenetic investigations. Methods. Analysis of BM cells chromosomal abnormalities in relapsed/refractory of MM for 62 patients with MM was performed. Slides of metaphase chromosomes were prepared according to standard procedure after cultivating during 24 or 96 h in the medium without stimulation. Results. Clonal chromosomal abnormalities were formed, complexity structure of karyotypes with clone structures was shown. In structural rearrangements, derivative chromosomes (34.9%) and balanced translocations (23.2%) were dominated at cytogenetic procedure. By molecular cytogenetic (i-FISH) investigation chromosomal abnormalities were detected in 26.7% cases. Del(17)(p13.1) was registered in 22.0%. Aberrations involving IGH were fixed in 5.7%. The atypical signal distribution determined by i-FISH may indicate another possible mechanism for the evolution of an abnormal clone. Conclusions. The cytogenetic peculiarities of the formation of abnormal BM cell clones in relapse/refractory of MM formations, discovered by us, supplement the understanding of the biological features of the disease and contribute to the outline of further tasks for the study of genetic aberrations in MM.

Poor prognosis of chromosome 7 clonal aberrations in Philadelphia-negative metaphases and relevance of potential underlying myelodysplastic features in chronic myeloid leukemia.

Clonal chromosome abnormalities in Philadelphia-negative cells could concern chronic myeloid leukemia patients treated by tyrosine kinase inhibitors. The European LeukemiaNet distinguishes -7/del(7q) abnormalities as a “warning”. However, the impact of clonal chromosome abnormalities, and specifically those of -7/del(7q), in Philadelphia-negative cells on clinical outcomes is unclear and based on case-reports showing morphological dysplasia and increased risk of acute myeloid leukemia, suggesting the coexistence of chronic myeloid leukemia and high-risk myelodysplastic syndrome. The aim of this study was to determine whether the impact of -7/del(7q) clonal chromosome abnormalities in Philadelphia-negative cells on the clinical outcome is different from that of other types of abnormalities, and we argue for an underlying associated high-risk myelodysplastic syndrome. Among 102 chronic myeloid leukemia patients with clonal chromosome abnormalities in Philadelphia-negative cells with more than a median of 6 years of follow up, patients with -7/del(7q) more frequently had signs of dysplasia, a lower cumulative incidence of deep molecular response and often needed further treatment lines, with the consequent impact on event-free and progression-free survival. Morphological features of dysplasia are associated with myelodysplastic syndrome/acute myeloid leukemia mutations and compromise the optimal response to tyrosine kinase inhibitors, irrespectively of the type of clonal chromosome abnormalities in Philadelphia-negative cells. However, mutation patterns determined by next-generation sequencing could not clearly explain the underlying high-risk disease. We hereby confirm the pejorative prognostic value of -7/del(7q) clonal chromosome abnormalities in Philadelphia-negative cells and suggest that myelodysplastic features constitute a warning signal that response to tyrosine kinase inhibitors may be less than optimal.

Inflammatory myofibroblastic tumour of gall bladder and liver: a case series & review of literature

Inflammatory myofibroblastic tumors are rare benign tumors that can mimic malignancy of unknown aetiology. It has spectrum of myofibroblastic proliferation along with varying amount of inflammatory infiltrate. Recently, the concept of this lesion being reactive has been challenged based on the clinical demonstration of recurrences and metastasis and cytogenetic evidence of acquired clonal chromosomal abnormalities. We hereby report a case series of three patients with inflammatory myofibroblastic tumour involving gallbladder, liver. Inflammatory myofibroblastic tumor (IMT) occurs more frequently in childhood and the most common involvement is seen in the lungs. Primary inflammatory myofibroblastic tumors of the gallbladder are rather infrequent. The present knowledge is based on case reports. Initiating factors such as reactive, infections, autoimmune and neoplastic processes, has been proposed but the etiology of most remains unknown. The most frequently involved organ in the abdomen is the liver, while primary gallbladder involvement is quite rare. Present knowledge about this entity is based on case reports in the literature (9). The clinical picture in cases with IMT depends on the organ of involvement and site of the organ; abdominal pain, jaundice and ascites might be the presenting symptoms, according to the localisation in the liver involvement. In our cases symptoms were abdominal pain and dyspepsia. CT demonstrated involvement of liver, gallbladder, which is rare. Hence, decision of extended cholecystectomy taken to do in these cases.

Abstract 3415: Cytogenetics of renal oncocytomas identify three distinct and mutually exclusive diagnostic classes of chromosome aberrations

Abstract Christopher B. Anderson1, Michael Lipsky1, Subhadra V. Nandula2,3, Christopher E. Freeman2, Thomas Matthews2, Catlin Walsh2, Gen Li1, Matthias Szabolcs2, Mahesh M. Mansukhani 2, James M. McKiernan1, Vundavalli V. Murty 2 1Departments of Urology, 2Pathology and Cell Biology, Columbia University Medical Center, New York, New York; 3Cancer Genetics Inc, Rutherford, New Jersey The cytogenetic alterations in renal oncocytoma (RO) are poorly understood. We analyzed 130 consecutive RO for karyotypic alterations. Clonal chromosome abnormalities were identified in 63 (48.5%) cases, which could be categorized into 3 classes of mutually exclusive cytogenetic categories. Class 1 RO had diploid karyotypes with 11q13 rearrangement in balanced translocations with ten or more different chromosome partners in all 20 (31.7%) cases. We identified recurrent translocation partners at 5q35, 6p21, 9p24, 11p13-14 and 11q23, and confirmed that CCND1 gene rearrangement at 11q13 utilizing fluorescence in situ hybridization (FISH). Class 2 RO exhibited hypodiploid karyotypes with loss of chromosome 1 and concurrent losses of Y in males and X in females in 25 (39.7%) cases. The class 3 tumors comprising of 18 (30.2%) cases showed diverse types of abnormalities with the involvement of 2 or more chromosomes exclusive of abnormalities seen in Class 1 and 2 tumors. Furthermore, karyotypically uninformative cases were subjected to FISH analysis to identify class 1 and 2 abnormalities. In this group we found similar frequencies of CCND1 rearrangement, loss of 1 or Y as with karyotypically abnormal cases. We validated our results against 91 tumors from the Mittleman database. Correlation of clinical data with all the 3 classes of RO showed no clear evidence of overall patient survival. Our findings support the hypothesis that RO exhibit 3 principal cytogenetic categories, which may have different roles in initiation and/or progression. These cytogenetic markers provide a key tool in the diagnostic evaluation of RO. Citation Format: Christopher B. Anderson, Michael Lipsky, Subhadra V. Nandula, Freeman E. Christopher, Matthews Thomas, Caitlin E. Walsh, Gen Li, Matthias Szabolcs, Mahesh M. Mansukhani, James M. McKiernan, Murty V. Vundavalli. Cytogenetics of renal oncocytomas identify three distinct and mutually exclusive diagnostic classes of chromosome aberrations [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 3415.

Cytogenetic features in primary myelodysplastic syndrome Egyptian patients

Karyotype is the most important diagnostic and prognostic parameter in myelodysplastic syndromes (MDS) and is abnormal in approximately 50% of patients. We emphasized the importance of chromosomal analysis and reported the most frequent cytogenetic abnormalities in 50 MDS (29 males (58%) and 21 females (42%), median age: 57.5 years) Egyptian patients using conventional banding analysis (CBA). Karyotype description was conducted according to the International System for Human Cytogenetic Nomenclature (ISCN, 2013). Patients were diagnosed based on complete history, bone marrow (BM) aspirate, peripheral blood (PBL) examination, and Iron stain. MDS with multilineage dysplasia (MDS-MLD) was the most frequently encountered subtype; 19/50 (38%) followed by MDS with single lineage dysplasia (MDS-SLD); 11/50 (22%). 27/50 patients (54%) showed a normal karyotype while 23 patients (46%) showed clonal nonrandom chromosomal abnormalities. Most patients with MDS with excess blasts-II (MDS-EB-II) showed abnormal karyotype (3/4; 75%) followed by MDS-EB-I (3/5, 60%) and MDS-MLD (10/19, 53%). Among 50 primary MDS patients; 14/50 (28%) had a single chromosomal abnormality, 3/50 (6%) had double chromosomal abnormality, and 6/50 (12%) had complex karyotype. Male sex was more frequently associated with higher IPSS prognostic risk categories than female gender. The most common single chromosomal abnormalities were −5/del5q; 7/50 (14%) patients followed by −7; 4/50 (8%) patients. +8, del20q and delY were each detected in 1/50 patient (2%). Abnormalities of chromosome 5 (−5/del5q) as a single chromosomal abnormality was the most frequent chromosomal abnormality among Egyptian primary MDS patients followed by complex karyotype. Cytogenetic characteristics of MDS Egyptian patients were similar to North African and European patients. Karyotype offers useful information in establishing accurate diagnosis and male gender is an important predisposing factor that can predict worse prognosis in MDS patients.

Wilms’ tumor gene 1 expression can predict sudden disease progression to blast crisis in patients with chronic myeloid leukemia receiving imatinib therapy

Background The Philadelphia chromosome is the hallmark of chronic myeloid leukemia (CML). As the disease progresses to accelerated phase and blast crisis (BC), clonal evolution may occur with the emergence of additional chromosomal abnormalities. Wilms’ tumor 1 gene (WT1) plays an important role in leukemogenesis, and its expression could represent a useful molecular marker of hematological malignancies. Aim The aim of this study was to evaluate WT1 expression and compare its kinetics with that of BCR-ABL1 expression in peripheral blood of patients with CML, to explore the utility of WT1 as an alternative marker for prediction of early disease progression and sudden transformation to BC. Patients and Methods A total of 49 newly diagnosed patients with CML, and 20 normal individuals as controls were enrolled in this study. WT1 and BCR-ABL1 expression was evaluated by quantitative real-time PCR. Results There was significant correlation between the expression of WT1 and BCR-ABL1 only at diagnosis and after 3 and 6 months of therapy. At 12 months of follow-up, an increase in %WT1 associated with low BCR-ABL1 was noticed in two of cases. One case suddenly progressed to BC at 18 months, and the other case showed emergence of clonal chromosome abnormality (+8) and progressed to accelerated phase. Conclusion Our results suggested that serial assessment of WT1 transcript level in patients with CML may be a useful marker for predicting early and sudden disease progression into advanced stages.

Clonal chromosomal abnormalities appearing in Philadelphia chromosome–negative metaphases during CML treatment

AbstractClonal chromosomal abnormalities in Philadelphia chromosome-negative (CCA/Ph−) metaphases emerge as patients with chronic phase chronic myeloid leukemia (CP-CML) are treated with tyrosine kinase inhibitors (TKIs). We assessed the characteristics and prognostic impact of 598 patients with CP-CML treated on clinical trials with various TKIs. CCA/Ph− occurred in 58 patients (10%); the most common were −Y in 25 (43%) and trisomy 8 in 7 patients (12%). Response to TKI therapy was similar for patients with CCA/Ph− and those without additional chromosomal abnormalities (ACAs). We further categorized CCA/Ph− into those in which –Y was the only clonal abnormality, and all others. We found that patients with non –Y CCA/Ph− had worse failure-free survival (FFS), event-free survival (EFS), transformation-free survival (TFS), and overall survival (OS) compared with those without ACAs with the following 5-year rates: FFS (52% vs 70%, P = .02), EFS (68% vs 86%, P = .02), TFS (76% vs 94%, P < .01), and OS (79% vs 94%, P = .03). In a multivariate analysis, non –Y CCA/Ph− increased the risk of transformation or death when baseline characteristics were considered with a hazard ratio of 2.81 (95% confidence interval, 1.15-6.89; P = .02). However, this prognostic impact was not statistically significant when achieving BCR-ABL <10% at 3 months was included in the analysis. In conclusion, non –Y CCA/Ph− are associated with decreased survival when emerging in patients with chronic-phase CML across various TKIs. This trial was registered at www.clinicaltrials.gov as #NCT00048672, #NCT00038649, and #NCT00050531 (imatinib); #NCT00254423 (dasatinib); #NCT00129740 (nilotinib); and NCT01570868 (ponatinib).

Cytogenetic and molecular cytogenetic investigations in relapse of B-cell chronic lymphocytic leukemia

Сhromosomal abnormalities have been analyzed in bone marrow cells of 61 patients with relapse of B-cell chronic lymphocytic leukemia. The cytogenetic results have allowed the structural stratification of the obtained karyotypes into ten groups of clones: normal, normal/near tetraploid, abnormal/normal, abnormal/ near tetraploid/normal, evolution of clonal chromosome abnormalities; evolution of clonal chromosome abnormalities/normal, evolution of clonal chromosome abnormalities/near tetraploid/normal, independent clones, independent/normal clones; and independent/near tetraploid/normal clones. The identified structural rearrangements included translocations, deletions, insertions, and duplications; however, deletions with the involvement of bands 17p12, 13q12–q14, 11q14, and 11q23 dominated (63.8%). The application of i-FISH helped to show the presence of one to four abnormalities per karyotype. The identified cytogenetic and molecular cytogenetic rearrangements may signify a multilevel nature of the process underlying the development of resistant karyotypes. The results obtained under both methods have revealed the presence of a heterogenic cell population with possibly different levels of chemotherapy resistance.

Risk Factors Associated with Survival Outcome in Patients with Chronic Myeloid Leukaemia in Accelerated Phase Treated with Tyrosine Kinase Inhibitors

Chronic Myeloid Leukaemia (CML) is a triphasic disease which typically presents in chronic phase with risk of progression to more aggressive phases in a certain proportion of patients. Accelerated Phase (AP), as described in the pre-Tyrosine Kinase Inhibitor (TKI) era by Kantarjian et al in 1988, is an intermediate stage with a poor median overall survival (OS) of ≤18 months without haematopoietic stem cell transplantation (HSCT). Since TKI therapy has revolutionized CML treatment, a significantly improved OS has been seen in most CML patients, including those in AP. Not all CML-AP patients require HSCT upfront nowadays and many are able to achieve major molecular remission (MMR) and favourable OS on TKI therapy. However, updated classifications of CML-AP by the World Health Organization (WHO) and European LeukemiaNet (ELN) do not reflect these significant advances in the TKI era. There is a need to re-evaluate the CML-AP classification that will have an impact on treatment decisions for CML-AP patients. In this study, we explored the association between various haematological parameters at diagnosis and the probabilities of OS and progression-free survival (PFS) of CML-AP patients on TKI therapy.Overall Survival (OS) and Progression-Free Survival (PFS) trends of 75 newly diagnosed CML-AP patients treated with frontline TKIs between 2000 to 2013 from Singapore General Hospital and Seoul St. Mary's Hospital in South Korea were retrospectively analysed with regards to demographic and haematological parameters, such as cell counts from serum and bone marrow at diagnosis, using cox proportional hazards analysis. Survival was also compared using log-rank test with Bonferroni corrections between CML-AP patients and 227 CML Chronic Phase (CML-CP) high-risk Sokal and 34 Blast Crisis (CML-BC) patients on TKI-based therapy. OS was defined as duration from diagnosis of CML-AP to death from any reason. PFS was defined as duration from disease diagnosis to the first occurrence of progression or death due to CML.As a whole, CML-AP patients treated with frontline TKI had survival that paralleled CML-CP high-risk Sokal patients (p-value = 0.694 for OS, p-value = 0.258 for PFS). Most of the death and progression occurred less than 3 years of starting TKI therapy (69.2% for OS, 84.6% for PFS). Multivariable analysis in CML-AP patients showed that male gender, bone marrow (BM) blasts ≥10% and clonal chromosomal abnormalities (CCAs) at diagnosis were associated with poor OS (Hazard Ratio (HR) 18.53, p-value = 0.013; HR 1.16, p-value = 0.010; HR 5.05, p-value = 0.044, respectively) and poor PFS (HR 12.96, p-value = 0.021; HR 1.17, p-value = 0.007; HR 8.84.05, p-value = 0.008, respectively). CML-AP patients with all 3 of these risk factors experienced the worst OS compared to those with 1 or zero risk factors (p-value <0.001). Patients with all 3 risk factors also had the poorest PFS compared to those with 2, 1 and zero risk factors (p-value = 0.022, <0.001, <0.001 respectively; figure 1). CML-AP Patients with 2 risk factors or less, had OS and PFS probabilities comparable to CML-CP patients with high-risk Sokal score (p-value = 0.082 for OS, p-value= 0.813 for PFS, figure 2 and 3 respectively). However, CML-AP patients with all 3 risk factors showed inferior OS and PFS probabilities similar to CML-BC patients (p-value = 0.799 for OS, p-value = 0.624 for PFS; figure 2 and 3 respectively).Our findings suggested that CML-AP was a heterogeneous group with varying survival probabilities on TKI therapy. Male gender, BM blasts ≥10% and CCAs at diagnosis were risk factors shown to be predictive of survival probabilities, and identified a high-risk sub-group among CML-AP patients with inferior OS and PFS rates similar to CML-BC patients. Aggressive chemotherapeutic strategies including HSCT should be warranted in these patients. However, TKI therapy alone with close molecular surveillance may be a reasonable option for optimally responding low-risk CML-AP patients who are not eligible for HSCT. [Display omitted] [Display omitted] [Display omitted] DisclosuresKim:BMS: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding, Speakers Bureau; Novartis: Consultancy, Honoraria, Research Funding, Speakers Bureau; ILYANG: Consultancy, Honoraria, Research Funding; Pfizer: Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding, Speakers Bureau. Goh:BMS: Honoraria; Novartis: Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding; Roche: Honoraria; Janssen: Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding; Celgene: Honoraria; Takeda: Honoraria; Alexion: Honoraria, Membership on an entity's Board of Directors or advisory committees.

CpG-Stimulated Cytogenetic Abnormalities Associated with Shorter Time-to-First Treatment

BackgroundPrognostic factors correlate with clinical outcomes independent of treatment. Many prognostic factors have been identified in CLL. More effective treatments, like B cell receptor (BCR) signaling pathway inhibitors, have the potential to nullify the prognostic impact of some markers. Time-to-first treatment is an endpoint unaffected by choice of treatment. CpG-stimulated metaphase karyotype can reliably and reproducibly identify cytogenetic abnormalities in CLL that may not be seen with standard non-stimulated karyotype or by FISH. Complex cytogenetics, defined as 3 or more chromosome abnormalities identified in 2 or more metaphases was the highest-risk feature for shorter progression-free and overall survival in patients receiving ibrutinib for relapsed/refractory CLL. Complex karyotype is not uncommon among relapsed/refractory CLL cases, particularly those who previously received genotoxic chemotherapy. The frequency and impact of karyotype abnormalities have not previously been reported for treatment-naïve CLL.MethodsWe evaluated treatment-naïve patients with CLL who had their initial evaluation, which included prognostic factor assessment, at MDACC between July 2013 and June 2016. CpG-stimulated metaphase karyotype of CLL cells from blood or bone marrow was performed by culture of mononuclear cells for 72hrs in media containing CpG-685 (20ug/ml), phorbol 12-myristate 13-acetate (PMA; 0.04ug/ml) and Pokeweed mitogen (PWM; 0.1ug/ml). Banding and analyses were by standard laboratory procedures. Twenty metaphases were analyzed per culture and patients were categorized as having diploid karyotype, a single clonal chromosome abnormality present in more than 1 metaphase, two clonal chromosome abnormities present in more than 1 metaphase, or 3 or more chromosome abnormalities identified in more than 1 metaphase (complex).ResultsCpG-stimulated karyotype was obtained in 500 treatment-naïve patients with CLL. The frequency and distribution of chromosome abnormalities with other prognostic factors and time-to-first treatment were analyzed (Table). The majority (69%) of patients had diploid cytogenetics. Higher-risk prognostic features such as del(17), del(11q), unmutated IGHV and ZAP70 expression were associated with presence of complex karyotype abnormalities. Shorter time-to-first treatment from diagnosis was associated with 1, 2, and 3 or more clonal chromosome abnormalities compared to diploid karyotype (p=.0005) (Figure).A previous multivariable model identified the following independent characteristics correlated with time-to-first treatment: size of the largest cervical lymph node, number of nodal sites involved, FISH cytogenetics, LDH, and IGHV mutation status (Wierda et al. JCO 29:4088, 2011). We are working to integrate stimulated cytogenetics results into this multivariable model for time-to-first treatment.ConclusionsClonal chromosome abnormalities were identified in nearly 30% of previously untreated patients with CLL and were associated with shorter time-to-first treatment. Analyses are ongoing to determine optimal integration into a multivariable model for time-to-first treatment. [Display omitted] [Display omitted] DisclosuresJain:Celgene: Research Funding; Novimmune: Consultancy, Honoraria; Genentech: Research Funding; Seattle Genetics: Research Funding; ADC Therapeutics: Consultancy, Honoraria, Research Funding; Incyte: Research Funding; Infinity: Research Funding; Novartis: Consultancy, Honoraria; Abbvie: Research Funding; Pharmacyclics: Consultancy, Honoraria, Research Funding; Servier: Consultancy, Honoraria; Pfizer: Consultancy, Honoraria, Research Funding; BMS: Research Funding. Thompson:Pharmacyclics: Consultancy, Honoraria. Burger:Gilead: Research Funding; Roche: Other: Travel, Accommodations, Expenses; Janssen: Consultancy, Other: Travel, Accommodations, Expenses; Portola: Consultancy; Pharmacyclics, LLC, an AbbVie Company: Research Funding. O'Brien:Pharmacyclics, LLC, an AbbVie Company: Consultancy, Honoraria, Research Funding; Janssen: Consultancy, Honoraria. Wierda:Genentech: Research Funding; Gilead: Research Funding; Acerta: Research Funding; Abbvie: Research Funding; Novartis: Research Funding.

Prognostic Value of BCR-ABL1 Transcript Type in Chronic Myeloid Leukemia Patients Treated Frontline with Nilotinib

Background. The fusion protein encoded by the BCR-ABL1 fusion gene may differ in size, but the great majority of chronic myeloid leukemia (CML) patients have a e13a2 (b2a2) or a e14a2 (b3a2) junction. In CML patients treated frontline with imatinib, the e14a2 transcript has been recently associated to faster and deeper molecular responses; in some studies a better outcome has been also reported. Very limited information on the prognostic impact of the BCR-ABL1 transcript type in CML patients treated frontline with second generation tyrosine kinase inhibitors (TKIs) is still available: a study from MDACC reported lower molecular response rates and a trend for inferior event-free survival in e13a2 patients.Aim. To evaluate if the BCR-ABL1 transcript type (e14a2 vs e13a2) affect the response and the clinical outcome in newly diagnosed adult CML patients treated frontline with nilotinib (NIL).Methods. An analysis of 345 CML patients in early chronic phase (ECP) enrolled within 3 multicentric prospective studies of the GIMEMA CML Working Party (ClinicalTrials.gov NCT00481052, NCT00769327, NCT01535391) was performed. The initial treatment was NIL 300 mg BID or NIL 400 mg BID. Definitions: major molecular response (MMR), BCR-ABL1IS ratio < 0.1%; deep molecular response (MR4.0), BCR-ABL1IS ratio < 0.01% with > 10,000 ABL1 copies; progression, transformation to advanced phases; death, at any time and for any reason. Cumulative incidences of response were estimated under consideration of competing risks (progression, death) and compared by Gray test. Progression-free survival (PFS) and overall survival (OS) were estimated using the Kaplan-Meier method and compared by log-rank test.Results. Seven patients expressing rare transcripts (e1a2 or e19a2) and 10 patients with unknown transcript type were excluded: 328 out of 345 patients were evaluable, 124 (38%) with e13a2 transcript, 174 (53%) with e14a2 transcript and 30 (9%) expressing both transcripts. The baseline characteristics of patients with e13a2 or e14a2 transcripts were comparable: no significant differences in age, gender, Sokal or EUTOS long-term survival score distribution, presence of clonal chromosomal abnormalities in Ph+ cells, NIL dose were observed; the only difference was a higher platelet count in patients with e14a2 transcript (median 374 vs 313 x 103/µl, p=0.006). The median follow-up was 60 months in both groups (range 24-82 months). The response rates and the survival probabilities were uniformly lower in patients with e13a2 transcript (N=124) compared to patients with e14a2 transcript (N=174), but the differences were not significant: MMR by 12 months, 66% vs 72%, p=0.244; MR4.0 by 36 months, 56% vs 66%, p=0.067; estimated cumulative incidence of MMR, 82% vs 88%, p=0.135; estimated cumulative incidence of MR4.0, 60% vs 69%, p=0.101; estimated PFS, 88% vs 93%, p=0.547; estimated OS, 89% vs 94%, p=0.436 (Figure 1). The responses and the survival probabilities of patients co-expressing the e13a2 and the e14a2 transcripts (N=30) were similar to or even better than the ones of e14a2 patients. Grouping together the patients with e14a2 transcript alone and the patients with co-expression of both transcripts (N=174+30=204), and comparing them to patients with e13a2 transcript alone (N=124), the response differences became significant (cumulative incidence of MMR and MR4.0, p=0.050 and p=0.036, respectively), but no outcome differences emerged (PFS and OS, p=0.340 and p=0.276, respectively).Conclusions. Despite a trend for lower response rates and inferior outcome in patients with e13a2 transcript, the observed differences were small and mostly not significant. Further studies in larger patient cohorts are required to clarify whether nilotinib and other second generation TKIs are able to overcome the adverse prognostic impact of transcript type, potentially affecting the speed and the depth of molecular response, the probability of achieving a treatment-free remission and the patient outcome. [Display omitted] DisclosuresCastagnetti:Bristol-Myers Squibb: Consultancy, Honoraria; Pfizer: Consultancy, Honoraria; ARIAD Pharmaceuticals: Consultancy, Honoraria; Novartis: Consultancy, Honoraria. Gugliotta:Bristol Myers Squibb: Consultancy, Honoraria; Novartis: Consultancy, Honoraria. Breccia:Ariad: Honoraria; Bristol Myers Squibb: Honoraria; Novartis: Consultancy, Honoraria; Pfizer: Honoraria; Celgene: Honoraria. Carella:Millenium: Speakers Bureau; Genentech: Speakers Bureau. Tiribelli:Ariad Pharmaceuticals: Consultancy, Speakers Bureau; Novartis: Consultancy, Speakers Bureau; Bristol-Myers Squibb: Consultancy, Speakers Bureau. Bocchia:Janssen: Honoraria; Novartis: Honoraria; Bristol-Myers Squibb: Honoraria. Soverini:Ariad: Consultancy; Bristol-Myers Squibb: Consultancy; Novartis: Consultancy. Cavo:Bristol-Myers Squibb: Consultancy, Honoraria; Millennium: Consultancy, Honoraria; Amgen: Consultancy, Honoraria; Janssen-Cilag: Consultancy, Honoraria; Celgene: Consultancy, Honoraria. Martinelli:ARIAD: Consultancy; Roche: Consultancy; MSD: Consultancy; Novartis: Speakers Bureau; Genentech: Consultancy; Amgen: Consultancy; BMS: Speakers Bureau; Pfizer: Consultancy, Speakers Bureau; Pfizer: Consultancy, Speakers Bureau; ARIAD: Consultancy; Roche: Consultancy; MSD: Consultancy; Amgen: Consultancy; Genentech: Consultancy. Rosti:Roche: Honoraria, Research Funding, Speakers Bureau; Incyte: Honoraria, Research Funding, Speakers Bureau; Pfizer: Honoraria, Research Funding, Speakers Bureau; BMS: Honoraria, Research Funding, Speakers Bureau; Novartis: Honoraria, Research Funding, Speakers Bureau.

The clinical features, cytogenetic characteristics and survival analysis of 550 myelodysplastic syndromes in a single center

目的回顾性分析550例初诊骨髓增生异常综合征(MDS)患者的临床特点、细胞遗传学特征和生存情况。方法收集患者基本信息，采用WHO(2008)标准进行分型，同时应用国际预后积分系统(IPSS)、基于WHO造血组织肿瘤分类的预后积分系统(WPSS)及修订版IPSS(IPSS-R)积分系统进行预后分层。结果550例MDS患者中位年龄57(12~89)岁，男女比为1.72∶1。血液学特点：外周血细胞三系可有不同程度减少或增加，中位HGB为72(22~154)g/L，中位PLT为52(3~587)×109/L，中位WBC为2.52(0.11~48.00)×109/L。细胞遗传学特征：251例(45.6%)检出克隆性染色体异常，复杂核型89例，占所有患者的16.2%。染色体异常类型主要以不平衡异常为主，三体或单体多见。最常见的克隆性异常是＋8，占染色体异常患者的29.9%(75/251)；其次为−7/del(7q)、del(5q)、del(20q)等。550例患者中位随访27(0.3~144)个月，采用IPSS、IPSS-R、WPSS分别对患者进行分组并比较各组生存，差异均有统计学意义(P<0.001)。其中IPSS-R分组中低危组中位OS期未达到，中危组中位OS期为44(95% CI 28~60)个月，高危组中位OS期为17(95% CI 13~21)个月，极高危组中位OS期为8(95% CI 5~11)个月(P <0.001)。结论该研究中纳入的MDS患者中位年龄57(12~89)岁，细胞遗传学异常类型与欧美报道有所差异，IPSS、IPSS-R和WPSS分层对于预测MDS患者的生存有重要意义。

Cytogenetic place in managing myelodysplastic syndromes: an update by the Groupe francophone de cytogénétique hématologique (GFCH).

The myelodysplastic syndromes (MDS) are preleukemic diseases of elderly patients characterized by defective maturation of clonal hematopoietic progenitor cells resulting in peripheral blood cytopenias. Clonal chromosomal abnormalities are heterogeneous and can be detected in less than 50% of patients with de novo MDS and more frequently in secondary MDS (up to 80%). The karyotype plays an important role in the pathogenesis, diagnosis, and prognosis to evaluate the risk of leukemic transformation and, more recently, in treatment allocation. The gold standard for cytogenetic diagnosis in MDS is conventional chromosome banding analyses of bone marrow metaphases. The most frequent abnormalities are deletions and losses of chromosomes 5 (-5/5q-) and 7 (-7/7q-) and various isolated or combined abnormalities. Fluorescent in situ hybridization and array comparative genomic hybridization can reveal cryptic genetic abnormalities but are not recommended in routine diagnosis. New techniques including next generation sequencing revealed somatic driver mutations especially those affecting genes involved in RNA splicing or those harboring important prognostic value (TP53, ASXL1…) with potential applications in clinical practice in the future.