Types Of Cytogenetic Abnormalities Research Articles

Tandem Autologous Stem Cell Transplantation Does Not Benefit High-Risk Myeloma Patients in the Maintenance Era: Real-World Results from The Canadian Myeloma Research Group Database

In patients with multiple myeloma, the presence of high-risk cytogenetic abnormalities is associated with worse disease control and survival. Autologous stem cell transplant does benefit these patients. Tandem transplantation has been explored as a means to deepen responses and further improve survival however, its role remains controversial. This is particularly true in the era of novel agent induction and post-transplant maintenance therapy. The aim of this study was to use the Canadian Myeloma Research Group database (CMRG-DB) and examine a large cohort of real-world patients comparing the outcomes of tandem versus single ASCT specifically in high-risk patients receiving novel agent-based induction and post-transplant maintenance. The data for this study was derived retrospectively from a comprehensive national-level database of Canadian patients with multiple myeloma. High-risk cytogenetics were defined as presence of del17p, t(4;14) or t(14;16). Those receiving allogeneic transplant were excluded. Tandem transplantation was defined as a second ASCT performed consecutively without interim relapse or progression after first ASCT. Those with relapse or progressive disease within three months of completing a first transplant were excluded. We compared response depth, progression free and overall survival based on single or tandem transplantation procedures. The impact of covariates of interest was also assessed. 381 patients with high-risk cytogenetics were identified. 242 received single and 139 patients received tandem transplants. All received post-transplant maintenance. The most common induction regimen for these patients was cyclophosphamide, bortezomib, and steroids (CyBorD, 87%). Forty-one patients (10.8%) required reinduction prior to first ASCT. The best overall responses at any time were 98.3% (90.5% ≥ VGPR) and 98.6% (89.9% ≥ VGPR) in the single and tandem ASCT groups respectively. Survival outcomes were similar with the median PFS for single or tandem ASCT of 35.2 and 35.3 months (p=0.88) and the median OS were 92.6 and 88.9 months respectively (p=0.72). No statistically significant differences were seen based on type of cytogenetic abnormality or type of maintenance. This was confirmed on multivariate analysis. In the real-world setting, tandem ASCT does not improve outcomes for MM patients with high-risk cytogenetics. This may be driven by the use of effective pre- and post-ASCT therapies. The development of more potent induction and consolidation along with current nearly ubiquitous continuous maintenance therapies until disease progression does not support the use of a second high dose procedure.

Molecular Cytogenetics and Immunophenotypic Characterization of Childhood Acute Leukaemia Patients in Tertiary Care Center of Bangladesh – An observative study

Background: Childhood leukemia is genetically a heterogeneous disease. Various types of cytogenetic abnormalities and immunophenotypic character are present in leukaemia which are important for risk stratification, treatment and play as significant prognostic factor. Pediatric acute leukaemia presents with varying clinical, morphological, immunological and molecular characteristics. It is very highly curable if diagnosed and treated properly. For detail typing and subtyping of acute leukemia immunophenotyping and cytogenetics are crucial. The aim of this study was to find out the genetic abnormalities and immunophenotypic characterization of childhood acute leukaemia patients in Bangladesh. Material & Methods: This was a retrospective observational study and was conducted in the Department of pediatric hemato-oncology of Combined Military Hospital, Dhaka and Ahsania Mission Cancer Hospital, Mirpur, Dhaka, Bangladesh during the period from February,2014 to March, 2022. There was total 98 cases of acute leukaemia. Results: In total 98 patients completed the study. We found that 79.59% patients were ALL and 20.41% patients were AML. Among ALL 80.64% were B cell type, 6.40% were T cell Type ; 12.82 % had TEL/AML1 or ETV6/RUNX1 t(12;21)(p13;q22), 5.13% patient had TCF3/PBX1 or E2A/PBX1 t(1;19)(q23;p13). In AML30.00% patient had PML/RARAt(15;17)(q22;q21), 10.00% patient had AML1/ETO or RUNX1/CBFA2T1 t(8;21) (q22;q22), 5.00% patient had FLT3/ ITD. In case of B-ALL highest expression of antigen was CD19 (91.64%) followed by CD10 (80.58%), HLADR (67.94%), CD22 (72.68%), CD79a (72.68%), TdT (52.14%) and CD34 (48.98%). In 44.24% cases there was co-expression of CD10 and CD19 and there was 11.6% expression of myeloid marker CD13 and 1.58% expression of T cell marker CD5. In case of T-ALL there was 100% expression of CD3. Expression of other antigen CD4, CD5, CD7, CD8, CD4/8 co-expression, TdT was 60%. There was 40% expression of CD1a and CD2. There was 20% expression of CD10, CD34 and TCRab also. In case of AML highest expression was MPO (93.75%) followed by CD33 (87.50%), CD13 (81.25%), CD117 (75%), HLADR (43.75%) and CD64 (50%). There was 6.25% aberrant expression of B-ALL marker CD19 and T-ALL marker CD3, CD4, CD5, CD7 also. Conclusion: Depending on this study we can say that except few variations distribution of immunophenotypical subtypes and genetic abnormalities of childhood acute leukaemia are almost similar to other literature published from neighboring countries.This study will serve as a guideline for future study in our country in this aspect.

SPECTRUM OF CYTOGENETIC ABNORMALITIES IN PEDIATRIC PATIENTS WITH ACUTE MYELOID LEUKEMIA

Objective: To determine the frequency and types of cytogenetic abnormalities in pediatric Acute Myeloid Leukemia. Material and Methods: This cross-sectional study was conducted from January 2021 to October 2021 in Hematology department of the CHUGHTAI Institute of Pathology in Lahore Pakistan. Total 60 patients who were newly diagnosed with Acute myeloid leukemia in CHUGHTAI Institute of Pathology and were also referred from Children Hospital Lahore were included. Patients under the age of 16 years and from both gender were included. Informed consent was taken from patient’s guardian/parents. Patients had their bone marrow aspirates processed for standard G-banding and their karyotypes were examined using a cyto-vision system, where the number of chromosomes was counted and examined for any changes, such as damaged, missing, rearranged, or additional copies of chromosomes. Morphology, immunophenotyping of aspirate, and trephine biopsy are used to make the diagnosis of AML. Data was entered in SPSS version 23.0. Result: There were total 60 patients included in the study. The mean age was 10.2+ 3.2 years. There were n=32(53.3%) males and n=28(46.6%) female. Normal karyotype was observed in n=36(60%) patients, abnormal were seen in n=14(23.3%) and unsuccessful was shown in n=10(16.6%) patients. Favorable cytogenetic results were seen in n=6 (10%), intermediate in n=41 (68.3%), and unfavorable in n=4 (6.6%) cases. t(8;21) (q22:q22) and t(15;17)(q24;q21) were found in 8.3% and 1.6% of our study population respectively which have favorable prognosis . One of our patients had complex cytogenetic abnormalities, including [del5p, del7q, del11q-17+] which shows poor prognosis. Conclusion: The study discovered that 14 (23.3%) of the total pediatric patients with acute myeloid leukemia showed aberrant cytogenetic abnormalities. Chromosomal abnormalities should be identified as early as possible since they can be used for AML risk stratification and prediction of prognosis. Keywords: Acute myeloid leukemia, Complex cytogenetic, Cyto-vision, Karyotype, Morphology, Pediatric

Cytogenetic abnormalities in a sample of females with premature ovarian failure

BackgroundPremature ovarian failure (POF) is a complex heterogeneous disorder characterized by the triad of amenorrhea, hypergonadotropinism, and hypoestrogenism in women before the expected age of menopause. In most POF patients, the etiology is idiopathic. X chromosome abnormalities are known to be responsible for many POF cases but the effect of sex chromosome low level mosaicism on ovarian function still remains unclear. The aim of this study was to investigate the prevalence and type of cytogenetic abnormalities as well as low-level sex chromosome mosaicism in Egyptian females with POF.ResultsThe present study recruited thirty women with POF and thirty women with normal reproductive history as a control group. Conventional cytogenetic analysis was carried out on POF patients in order to detect cytogenetic abnormalities. FISH on interphase and metaphase nuclei from patients with normal karyotype as well as from thirty control women with normal reproductive history was performed using X, Y, and 18 centromeric probes to evaluate low-level sex chromosome mosaicism. Conventional cytogenetic analysis of peripheral blood lymphocytes demonstrated chromosomal aberrations in 7 cases. FISH revealed that the rate of X chromosome mosaicism was significantly higher in POF patients than in the control group.ConclusionWe concluded that X chromosome abnormalities including low level mosaicism may be underlying the pathology of POF as increased mosaicism may lead to accelerated oocyte aging and premature follicular atresia.

P–383 Cytogenetic analysis of products of conception from miscarriages following natural conception and IVF pregnancy: a comparative study

Abstract Study question Do cytogenetic results from products of conception from miscarriages differ from patients conceiving by natural conception versus IVF? Summary answer: Cytogenetic results were similar, with no statistical difference from miscarriages following natural conception and assisted conception. What is known already Cytogenetic sampling of products of conception (POC) following recurrent miscarriages (RM) are recommended to rule out parental chromosomal rearrangements. The RCOG recommends cytogenetic testing in cases of recurrent miscarriages (3 consecutive miscarriages). However some units routinely request cytogenetic analysis following a single miscarriage following an IVF pregnancy. There is no evidence to support the routine sampling of POCs following assisted conception. Study design, size, duration: Retrospective cohort study of 117 cytogenetic samples, followed up by the outcome of parental karyotyping if applicable. Patients were categorised based on mode of conception into natural conception (NC) with recurrent miscarriages (≥3) or one miscarriage following IVF. Data collected between 2018–2020. Primary Outcome measure: Presence and type of cytogenetic abnormality; individual parental targeted G-band karyotyping result. Participants/materials, setting, methods: A total of 117 cytogenetic results were reviewed, of which 35 were unsuitable for analysis due to contamination (Total n = 79: NC = 60, IVF = 19). Main results and the role of chance: Cytogenetic analysis showed abnormal results in 59% of miscarriages following natural conception and 53% of miscarriages from IVF pregnancy (p = 0.46). Abnormal cytogenetic results were mainly sporadic. Trisomy 16 was the commonest abnormality in both groups. Others included Trisomy 15, 22, 21, 8, 13, 5, 9, 10, 14, 18, single X (Turner’s), all occurring in the same frequency in both groups. As expected 35 out of 45 abnormal cytogenetic results occurred with a maternal age greater than 35 years. One couple from the NC group were referred to a geneticist for a Trisomy 9 imbalance. All other parental karyotyping results were normal. Limitations, reasons for caution This study contains a small sample size, and would benefit from further data collection to account for a percentage of samples being inadequate for analysis. Wider implications of the findings: Cytogenetic results were similar from miscarriages following natural conception and assisted conception. IVF does not increase the risk of miscarriage from abnormal embryonic karyotype. Routine cytogenetic testing following one miscarriage in patients undergoing IVF is not cost effective. Trial registration number Not applicable

Adverse prognostic impact of complex karyotype (≥3 cytogenetic alterations) in adult T-cell acute lymphoblastic leukemia (T-ALL)

The potential prognostic value of conventional karyotyping in adult T-cell acute lymphoblastic leukemia (T-ALL) remains an open question. We hypothesized that a modified cytogenetic classification, based on the number and type of cytogenetic abnormalities, would allow the identification of high-risk adult T-ALL patients. Complex karyotype defined by the presence of ≥3 cytogenetic alterations identified T-ALL patients with poor prognosis in this study. Karyotypes with ≥3 abnormalities accounted for 16 % (22/139) of all evaluable karyotypes, corresponding to the largest poor prognosis cytogenetic subgroup of T-ALL identified so far. Patients carrying karyotypes with ≥3 cytogenetic alterations showed a significantly inferior response to therapy, and a poor outcome in terms of event-free survival (EFS), overall survival (OS) and cumulative incidence of relapse (CIR), independently of other baseline characteristics and the end-induction minimal residual disease (MRD) level. Additional molecular analyses of patients carrying ≥3 cytogenetic alterations showed a unique molecular profile that could contribute to understand the underlying molecular mechanisms of resistance and to evaluate novel targeted therapies (e.g. IL7R directed) with potential impact on outcome of adult T-ALL patients.

Types of Cytogenetic Abnormalities in the Apical Root Meristem of Elodea canadensis from the Yenisei River Areas with Different Types of Anthropogenic Pollution

The change in the spectrum of chromosomal abnormalities in the apical root meristem of Elodea canadensis Michx. (elodea) growing in areas with different types of anthropogenic (radioactive and chemical) pollution of the bottom sediments of the Yenisei River is shown. In the samples of elodea collected in the zone of radioactive contamination of the river by discharges of the Rosatom Mining and Chemical Combine (MCC, Zheleznogorsk), the predominant type of abnormalities consist of chromosomal aberrations (bridges, stickiness, chromosomal fragmentation), as well as multiple abnormalities in one cell. The elodea samples from the areas of the Yenisei River with nonradioactive (chemical) contamination of bottom sediments by the enterprises of the city of Krasnoyarsk are predominantly characterized by abnormalities associated with the disturbance of the mitotic spindle of cell division.

Types of cytogenetic abnormalities in apical root meristem of Elodea сanadensis from Yenisei River areas with different types of anthropogenic pollution

Background. The spectrum of cytogenetic abnormalities in the apical root meristem of Elodea сanadensis Michx. (elodea) from the Yenisei River was studied depending on the type of anthropogenic pollution. We compared elodea samples from areas with radiation-contaminated sediments (area affected by Rosatom Mining and Chemical Combine, MCC, Zheleznogorsk) and areas with chemical pollution (Krasnoyarsk). Materials and methods. The apical root meristem of elodea was fixed in acetic alcohol (96 % ethanol and glacial acetic acid, 3 : 1) and stained with 1 % aceto-orcein. The cytogenetic analysis of metaphase and ana-telophase elodea cells was carried out with temporal squashed preparations using an Olympus CX31 microscope. Results. At metaphase stage, the predominant types of abnormalities in elodea cells were disoriented chromosomes (up to 9 % of total metaphase cells) and chromosome agglutination (6 %). In the area affected by the MCC an increased content of ring chromosomes in elodea cells was detected, but in terms of frequency of their occurrence no significant differences were revealed between samples from areas with different types of pollution. Among abnormalities at ana-telophase stage, bridges (to 20% of dividing cells) and disoriented chromosomes (up to 8%) dominated. The following abnormalities were also detected: multipolar and asymmetrical mitoses, agglutination and cells with multiple chromosome abnormalities. It was shown that in areas with the highest content of 137Cs in bottom sediments the frequency of cells with bridges and multiple chromosome abnormalities significantly increased as compared to samples from other areas. Conclusion. In the radioactive contamination area of the Yenisei River the spectrum of abnormalities was dominated by chromosome aberrations (bridges, agglutination, chromosome fragmentation) and multiple abnormalities in one and the same cell. In areas with non-radioactive (chemical) contamination of sediments, mitotic spindle irregularities prevailed.

The Epigenetic Repression of Mir-375 Is the Dominant Mechanism for the Constitutive Activation of PDPK1/RSK2 Signaling Axis in Multiple Myeloma

Abstract [Introduction] Multiple myeloma (MM) is a cytogenetically/molecularly heterogeneous hematologic malignancy that remains mostly incurable, and the identification of a universal and relevant therapeutic target molecule is essential for the further development of therapeutic strategy. We have recently identified that 3-phosphoinositide-dependent protein kinase 1 (PDPK1), a serine threonine kinase, and its major downstream substrate RSK2, a member of the 90 kDa ribosomal S6 kinase family of serine threonine kinases, were universally active in eleven human MM-derived cell lines (HMCLs) examined regardless of the type of cytogenetic abnormality, the mutation state of RAS, RAF and FGFR3 genes and myeloma cells of approximately 90% of symptomatic patients at diagnosis. Our study also disclosed that PDPK1/RSK2 signaling axis played pivotal roles in myeloma pathophysiology by regulating series of downstream molecules, such as c-MYC, IRF4, D-type cyclins, or PLK1, while the inactivation of either PDPK1 or N-terminal domain of RSK2 resulted in the induction of apoptosis in myeloma cells which was accompanied by the activation of BH3-only proteins BIM and BAD (Shimura Y, Mol Caner Ther 2012; Chinen Y, Cancer Res 2014). Here we assessed the underlying mechanism for PDPK1 overexpression in MM. [Methods] The miR-375 expression level was analyzed by the quantitative RT-PCR in 11 HMCLs and 92 patient-derived myeloma cells isolated by CD138-positive cell sorting (normal plasma cells (N=10), MGUS (N=21), newly diagnosed MM (NDMM) (N=27), relapsed/refractory (RRMM) (N=34). The pre-miR-375 precursor molecule (miR-375 mimics), the siRNA targeted against PDPK1, or a negative control RNA-oligonucleotides was transfected into 8 cell lines by utilizing Hemagglutinating Virus of Japan (HVJ)-envelope vector. The copy number abnormality of PDPK1 gene was assessed by double-color FISH for PDPK1 gene and the centromere of chromosome 16. The methylation status of miR-375 promoter site was analyzed by methylation-specific PCR (MSP). This study was conducted in accordance with the Declaration of Helsinki and with the approval of the Institutional Review Boards. Patient-derived samples were obtained with informed consent, and normal bone marrow plasma cells were obtained from volunteers who were not affected by hematologic disease. [Result] The level of miR-375 expression was calculated with 2-ΔCt methods. Human U6 snRNA was examined as the reference. The median log102-ΔCt ± SD of normal plasma cells, MGUS, NDMM, RRMM and HMCLs were -2.46 ± 0.67, -3.64 ± 0.68, -4.23 ± 0.95, -3.92 ± 1.24 and -3.69 ± 0.29 respectively. When compared to normal plasma cells, the miR-375 expression was significantly decreased in NDMM and RRMM (p<0.01, respectively), and tended to be decreased in MGUS (p=0.083) and HMCLs (p=0.097). As the causative of miR-375 repression, our study disclosed that the promoter sites of miR-375 gene were hypermethylated in 8/8 of HMCLs when examined by MSP. The interphase FISH for PDPK1 with centromere chromosome 16 indicated the copy number of PDPK1 gene was increased in 11/11 HMCLss, however, this was never the case with patient-derived myeloma cells (0/7). Importantly, the miR-375 gene transfection resulted in the reduction of PDPK1 expression in 7 of 8 HMCLs, and it simultaneously caused the reduction of the expression levels of IGF1 receptor and JAK2, the known targets of miR-375. Furthermore, when treated with 5-Azacitidine and/or Trichostatin A, miR-375 was markedly upregulated, suggesting that the overlapping epigenetic deregulations, such as DNA hypermethylation or histone deacetylation, are involved in the silencing of miR-375. [Conclusion and Discussion] PDPK1 is activated by autophosphorylation and, therefore, its expression level is the crucial determinant for its activity, Because our study revealed the miR-375 expression as the major regulator of PDPK1 expression, it is suggested that the abnormally repressed miR-375 is the major causative for the constitutive hyperactivation of the PDPK1/RSK2 signaling axis in MM. Moreover, since the decreased miR-375 expression was observed in plasma cells of MGUS and was more pronounced in MM, miR-375 repression by epigenetic deregulation may be involved in both disease development and progression of MM. Disclosures No relevant conflicts of interest to declare.

Conventional Cytogenetics and FISH [del13q, del17p, t(11;14), t(4;14)] Findings and Their Relationship with Other Risk Factors in Multiple Myeloma

Multiple myeloma (MM) represents a malignant proliferation of plasma cells in bone marrow. The prognosis is highly variable and mar- kedly affected by a number of prognostic features. We aimed to determine the relationship between conventional cytogenetics and FISH (del13q, del17p, t(11;14), t(4;14)) findings and other prognostic factors in MM. In our study, we investigated the frequencies and the types of cytogenetic abnormalities in 50 MM patients. The most common chromosomal abnormality was hypodiploidy ob- served in 40.5% of 42 patients (17/42) screened with conventional cytogenetic analysis. Del 13q was detected in 37.5% of 48 pa- tients (18/48)) screened with fluorescence in situ hybridization (FISH). 29 patients were investigated for del 17p, t(11;14), and t(4;14). Del 17p was detected in 3 (10.3%), t(11;14) in 7 (24.1%), t(4;14) in one (3.4%) of them. International Staging System (ISS) stage was increased as the Durie-Salmon (DS) stage increased. We didn't find a relationship between the cytogenetic abnormalities that we studied and DS or ISS stages.

Therapy-Related Myelodysplasia/Acute Myeloid Leukemia (t-MDS/AML) After Conventionally-Treated Breast Cancer: Impact of Cytogenetics on Outcome After Allogeneic Hematopoietic Cell Transplantation (HCT),

Abstract 4149 Background: The outcome of early stage breast cancer has improved in recent years with use of effective combination chemotherapy, including cyclophosphamide, anthracyclines or taxanes. Patients who undergo lumpectomy also receive radiation therapy for locoregional control. t-MDS/AML are rare but serious consequences of these therapeutic exposures; historically with an overall poor prognosis. Translocations involving the mixed-lineage leukemia (MLL) gene on chromosome band 11q23, and core binding factor genes on 21q22 and 16q22, are hallmarks of t-MDS/AML resulting from treatment with topoisomerase II inhibitors (anthracyclines). Loss of part or whole of chromosomes 5 and/or 7 (5-/7- abnormality) are associated with t-MDS/AML after treatment with alkylating agents and radiation. Allogeneic HCT (sibling-related, unrelated or double-cord) offers the only curative option for some of these patients. However, the long-term outcome of post-breast cancer t-MDS/AML after allogeneic HCT has not been described, and subgroups with better or worse prognosis not identified. Methods: From 1997 to 2011, 28 patients with t-MDS/AML after treatment of breast cancer with conventional chemotherapy and/or radiation underwent allogeneic HCT at City of Hope. Overall survival defined from the time of allogeneic HCT to the last date of follow up or date of death, was estimated using Kaplan-Meier method. Cox regression techniques were utilized to understand predictors of overall survival. Results: Of the 28 patients, 16 had been treated for early-stage breast cancer (I/IIA), 11 had received treatment for locally advanced disease (IIB/ III) and 1 patient had metastatic disease. Twenty-five patients received combination chemotherapy as adjuvant therapy or for metastatic disease; this included cyclophosphamide (100%), doxorubicin (88%) and taxanes (52%). Details of chemotherapy are unknown for one patient. Twenty-one patients (75%) also received radiation (2 received radiation alone). Median age at time of diagnosis of t-MDS/AML was 56 years (range: 34–79); median time from diagnosis of breast cancer to diagnosis of t-MDS/AML was 2.4 years (range 1–12.3); median time from t-MDS/AML to allogeneic HCT was 0.55 year (range: 0.15–4.1). Ten patients presented with MDS and 18 with AML. Cytogenetic abnormalities included MLL - 11q23 abnormality (n=12), 5-/7- and/or complex abnormalities (n=9), t(15;17)(q22;q21.1) (n=1), t(8;21) (n=1), inv(16)(p13.1;q22) (n=2). Of the 28 patients, 27 were treated with induction therapy and 74% were in complete remission (CR) at the time of HCT. Fourteen patients (50%) received sibling-related HCT, 13 received matched unrelated donor (MUD) transplant and 1 received a double cord transplant. Two patients received second allogeneic HCT and one patient received donor lymphocyte infusion (DLI) for relapsed t-MDS/AML. Sixty-one percent received reduced-intensity conditioning with fludarabine/melphalan and others received full-intensity conditioning. Sixteen patients developed cGvHD after transplant. After a median follow up of 2.2 years, 8 patients had died (cause of death: t-MDS/AML [n=5], sepsis [n=1], GvHD [n=1], breast cancer [n=1]), with an overall survival of 78.2% at two years post-HCT for the entire cohort. The 2-year overall survival was 71.4% for sibling-related HCT and 84.4% for MUD (log-rank p=0.14). There was a difference in overall survival by the type of cytogenetic abnormalities. Thus, the 2-year survival rate for patients with 11q23 abnormality was 90.9% while that for patients with chromosome 5-/7- abnormalities or complex abnormalities was 62.5% (log rank p=0.27). Patients with cGvHD had better outcome compared with those without cGvHD (2 year survival 87.5% vs. 64.3%, log-rank p value=0.03). These findings were confirmed on multivariate regression analysis, which revealed chromosome 5-/7- or complex abnormalities to be associated with a significantly worse outcome (HR=6.9, p=0.035), while cGvHD was associated with a better outcome (HR=0.14, p=0.02) after adjustment for age at HCT. Conclusions: Although t-MDS/AML after conventional treatment of breast cancer has historically been shown to have a poor prognosis, allogeneic HCT in patients with available donors provides a chance of cure. Among patients undergoing allogeneic HCT, a superior outcome was observed in patients with 11q23 abnormalities and among those with cGvHD. Disclosures: No relevant conflicts of interest to declare.

The risk of cytogenetic abnormalities in the late first trimester of pregnancies conceived through assisted reproduction

To determine if pregnancies conceived through infertility treatment are at increased risk of cytogenetic abnormalities in the late first trimester compared with spontaneously conceived pregnancies, or if there is increased risk when comparing less invasive infertility treatment (in vivo group) to in vitro fertilization (in vitro group). Retrospective case-controlled study. University hospital. A total of 1,606 women who spontaneously conceived and 559 women who conceived through infertility treatment undergoing chorionic villus sampling (CVS). None. Cytogenetic abnormalities diagnosed by CVS. No difference in cytogenetic abnormalities was found when comparing spontaneously conceived pregnancies to those conceived through infertility treatment (7.0% versus 5.4%). We also found no difference in the prevalence of cytogenetic abnormalities when comparing in vivo and in vitro fertilization subgroups (4.7% versus 5.8%). Finally, no difference was found when comparing the prevalence of different types of cytogenetic abnormalities between groups. Infertility treatment does not increase the risk of carrying a cytogenetically abnormal fetus in the late first trimester, nor does it increase the preponderance for any specific type of abnormality.

Chromosomal abnormalities in transformed Ph‐negative myeloproliferative neoplasms are associated to the transformation subtype and independent of JAK2 and the TET2 mutations

Evolution to myelofibrosis (MF), acute myeloid leukemia or myelodysplastic syndrome (AML/MDS) may occur over time in myeloproliferative neoplasms (MPN) patients most likely due to the acquisition of additional mutations. The Groupe Francophone de cytogenetique hematologique (GFCH) has collected and reviewed 82 patients with transformation of MPN (66 AML/MDS and 16 MF). JAK2V617F and TET2 mutations were searched for in 40 and 32 patients, respectively. Significantly more -7/del(7q) (P = 0.004) and -5/del(5q) (P = 0.03) were found in AML/MDS with a higher incidence of dup1q (P = 0.01) in MF. Some specific chromosomal abnormalities occurred together, for example -5/del(5q) and -17/del(17p) (P = 0.0007). In multivariate analysis, two factors were independently associated with an inferior overall survival (OS); AML/MDS transformation (P < 0.0001) and -5/del(5q) abnormality (P = 0.02). Although both giving rise to loss of 7q, der(1;7) differed from other 7q deletions in terms of distribution (lower frequency of AML/MDS, P = 0.02), association with chromosomal abnormalities (absence of -5/del(5q), P = 0.003; increased del(20q), P = 0.05), and longer OS (P = 0.0007). We detected 24/40 (60%) JAK2V617F and 8/25 (32%) TET2 mutations in samples following transformation, ranging from wild-type to mutated forms of both genes. The mutated and wild-type forms of the genes were not found to be associated with a specific chromosomal abnormality. There was no evidence that JAK2 or TET2 mutations were associated with the type of MPN transformation, whereas the type of cytogenetic abnormalities were strongly linked, perhaps indicating that they play a specific role in the transformation process.

Pediatric Myelodysplastic Syndromes and Allogeneic Hematopoietic Progenitor Cell Transplantation (HPCT): A Single Center Perspective.

Abstract 1780Poster Board I-806 IntroductionMyelodysplastic syndromes (MDS) in pediatrics comprise a heterogenous group of disorders that are poorly understood and difficult to treat. Allogeneic HPCT remains the only curative treatment, but published outcomes in children are limited. MethodsWe performed a retrospective review of pediatric patients with MDS who received an allogeneic HPCT on IRB approved protocols between 1992 and 2009 at Children's Memorial Hospital (Chicago, IL). ResultsThe study population consisted of 23 consecutive patients. Median age at transplant was 9.2 years (range 4.6-17.2 years) and median time to transplant from diagnosis was 4.3 months (range 2.2-9.9 months). Nine patients (39%) had de novo MDS, whereas 14 of 23 (61%) patients had treatment-related MDS, and 4 patients in this group had received a previous autologous transplant for a previous malignancy. All patients had a documented cytogenetic abnormality: monosomy 7/7q- in 12 patients (52%), trisomy 8 in 3 (13%), translocation (6;9) in 3 (13%), and others in 5 (22%). Donors were HLA matched siblings in 8 cases (35%), 8/8 loci HLA matched unrelated donors in 6 (26%), and the remaining 9 (39%) were 6-7/8 loci HLA mismatched unrelated donors. HPC source was peripheral blood in 13 patients (57%), cord blood in 7 (30%), and bone marrow in 3 (13%). Fourteen of 23 (61%) patients received a myeloablative conditioning regimen with TBI/Cy in 9 (31%), Bu/Cy in 4 (17%), and Flu/Bu/low-dose TBI in 1. Reduced intensity conditioning (RIC) regimens were used for the remaining 9 (39%) and consisted of Flu/Bu/ATG. Indications for RIC transplant included patient choice in 4 patients, prior autologous transplants in 3 patients, and comorbid conditions in 2 patients. Twenty patients (90%) had evidence of engraftment, with only 1 death prior to day 100. Seven patients (30%) died secondary to transplant-related complications, including infection in 4, GVHD in 2, and toxic epidermolysis in 1. Of the 4 patients that had received a prior transplant, 3 died from transplant-related mortality. Grade 2-4 acute and chronic graft-versus-host disease (GVHD) was seen in 9 (39%) and 11 (48%) patients, respectively. For the entire group, estimated five year relapse-free survival (RFS) and overall survivals (OS) were 47% [95% CI: 28%, 67%] and 50% [95% CI: 25.1%, 71.1%], respectively. Two patients had graft failure, which included 1 patient who received cord blood and 1 patient who received peripheral blood with RIC. Five patients suffered a relapse of original disease, including 3 patients who received RIC. These 3 patients each received a second myeloablative transplant; 2 of these patients remain alive and disease-free. Treatment-related MDS was associated with decreased RFS in comparison to non-treatment-related MDS (33% [95% CI: 11%, 58%]) vs. 70% [95% CI: 22%, 92%], p=0.05). In contrast, five year OS rates reached 80% for those with de novo MDS. There was no significant difference in overall survival between patients who received RIC versus myeloablative conditioning (p=0.17), but RIC regimens were associated with decreased relapse-free survival in comparison to myeloablative conditioning regimens (22% [95% CI: 3%, 51%] vs. 68% [95% CI: 34%, 87%], p=0.02). On univariate analysis, there was no correlation of relapse-free or overall survival with blast count at diagnosis, IPSS score, type of cytogenetic abnormality, receiving AML-like chemotherapy for MDS, CMV serostatus, donor type and HLA match, cell dose, and time to transplant. ConclusionsAllogeneic HPCT is a curative option for pediatric MDS. In this series of high risk patients we achieved OS approaching 80% in patients with de novo MDS. Those who received RIC had an increased relapse rate but were salvaged by myeloablative conditioning regimens and had comparable outcomes to the myeloablative group. The role of RIC regimen transplants in pediatric MDS needs to be studied further. Larger series of pediatric patients are needed to establish a prognostic scoring system to predict outcome and select appropriate conditioning regimens to further improve survival in allogeneic HPCT for MDS. [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

Ploidy and karyotype complexity are powerful prognostic indicators in the Ewing's sarcoma family of tumors: A study by the United Kingdom Cancer Cytogenetics and the Children's Cancer and Leukaemia Group

Ewing's sarcoma family tumors (ESFT) are characterized by the presence of EWSR1-ETS fusion genes. Secondary chromosome changes are frequently described, although their clinical significance is not clear. In this study, we have collected and reviewed abnormal karyotypes from 88 patients with primary ESFT and a rearrangement of 22q12. Secondary changes were identified in 80% (70/88) of tumors at diagnosis. Multivariate analysis showed a worse overall and relapse free survival (RFS) for those with a complex karyotype (overall survival, P = 0.005; RFS, P = 0.04), independent of metastatic disease. Univariate survival analysis showed that a chromosome number above 50 or a complex karyotype was associated with a worse overall survival (>50 chromosomes, P = 0.05; complex karyotype, P = 0.04). There was no association between type of cytogenetic abnormality and the presence of metastatic disease at diagnosis. Univariate and multivariate survival analysis of a small subgroup with trisomy 20 indicated that trisomy 20 was associated with a worse overall and RFS. There was no difference in outcome associated with other recurrent trisomies (2, 5, 7, 8, or 12) or the common recurrent secondary structural rearrangements (deletions of 1p36, 9p12, 17p13, and 16q, and gain of 1q), although numbers were small. These data demonstrate the continued value of cytogenetics as a genome-wide screen in ESFT and illustrates the potential importance of secondary chromosome changes for stratification of patients for risk. Specifically, karyotype complexity appears to be a powerful predictor of prognosis, and the presence of trisomy 20 may be a marker of a more aggressive subset of this group.

The Association between Cytogenetic Abnormalities and Clinical Outcomes Based on Prognostic Factors of the Children Cancer Group (CCG) in Pediatric Patients with Acute Leukemia: Two Institutional Retrospective Studies

Results: The incidence of cytogenetic abnormalities was 51 (51.5%) in 99 patients, and 27 (39.7%) in acute lymphoblastic leukemia (ALL) patients and 24 (77.4%) in acute myelogenous leukemia (AML) patients. The most frequent cytogenetic abnormality was hyperdiploidy and t(8:21) in the ALL and AML patients, respectively. The overall survival rate (OS)/disease free survival rate (DFS) of the ALL patients was 74.0%/73.9%. The OS/DFS of the standard risk group (88.8%/85.2%) was significantly higher than that of the high-risk group (49.4%/39.3%) in the ALL patients (P=0.0005/P<0.0001). There was no significant difference in the survival rates according to the type of cytogenetic abnormalities among the ALL patients for the standard/high risk groups, based on the CCG prognostic factors. The OS/DFS of the AML patients were 43.4% and 41.7%, respectively, without significant differences of the survival rates according to the type of chromosomal abnormalities. Conclusion: There were significant differences of OS/DFS based on the risk groups in ALL patients when evaluated with the CCG prognostic factors (standard/high) and chromosomal abnormalities (good/ poor), respectively. However, there was no significant correlation between type of cytogenetic abnor- malities and clinical outcomes based on the CCG prognostic factors in children with ALL as well as with AML.

Biological and clinical significance of clonogenic assays in patients with myelodysplastic syndromes.

Biological and clinical significance of growth pattern of hematopoietic progenitors were investigated in 117 patients with primary myelodysplastic syndromes (MDSs) at referral. Abnormal (i.e., "leukemic" or absent) growth of GM colonies (CFU-GM) and GM clusters was found in 47% of patients with "advanced" MDS (RAEB, RAEB-t, and CMML) and in 15% of "low-risk" (RA/RARS) patients. In vitro erythropoiesis was decreased in most of the patients, with significantly lower number of BFU-E in "advanced" MDS than in RA/RARS patients. Megakaryocyte progenitors (CFU-MK) were very low or absent in almost all the patients, regardless of the FAB type. Significant correlation was demonstrated between the number of BFU-E and hemoglobin concentration and between number of CFU-MK and platelet count. Growth capacity of GM progenitors appears to be in proportion to "myeloproliferative" capacity of the malignant clone. T-cell depletion had no influence on growth capacity of hematopoietic progenitors, nor did colony growth respond in a dose-dependent manner to different concentrations of LCM. Growth capacity of MDS hematopoietic progenitors was independent of Bournemouth score, of the presence and type of cytogenetic abnormality, and of the expression of CD95 and caspase-3 antigens on bone marrow cells. However, in patients with "abnormal" growth of GM progenitors, CD34 antigen expression was significantly higher than in patients with "normal" growth. "Abnormal" GM growth was found to be independently predictive regarding the survival and the risk for AML development. In contrast, the prognostic value of erythroid and megakaryocyte cultures was found to be limited.

Terminal Deoxynucleotidyl Transferase Expression in Acute Myelogenous Leukemia and Myelodysplasia as Determined by Flow Cytometry

The significance of terminal deoxynucleotidyl transferase (TdT) expression in acute myelogenous leukemia (AML) remains controversial. Therefore, we studied TdT expression by flow cytometry in 120 previously untreated patients with AML or myelodysplatic syndrome (MDS) to determine the distribution of TdT-positive blasts and the intensity of TdT expression and to seek clinically significant associations. TdT expression measured by flow cytometry (flow TdT%) was heterogeneous, ranging from 0.1% to 87% (median, 8.5%), and 74 patients (62%) had at least 5% TdT-positive blasts. TdT positivity was associated with the MO or Ml subtype and with expression of CD34 and CD7. No significant correlation was found between TdT expression and type of cytogenetic abnormality or rearrangement of immunoglobulin or T-cell receptor genes. Remission lasted longer in patients with a flow TdT% ≤ 5 than in patients with a flow TdT% ≥ 5 (median, 95 weeks vs 55 weeks, p = 0.02); however, complete remission rates did not differ when patients were classified by initial flow TdT%. Survival was slightly better for patients with flow TdT% less than 5% Among patients with a flow TdT% ≥ 5%, those with a higher TdT intensity survived longer than those with a lower intensity. These data suggest that quantitative TdT measurement may contribute to prognostic estimate in AML patients.