Major Chromosomal Aberrations Research Articles

High rates of "atypical" single nucleotide polymorphism-based noninvasive prenatal screening results among consanguineous Arab American patients: A single center retrospective study.

Noninvasive prenatal screening (NIPS), using placental cell-free DNA from a maternal blood sample, is currently the most sensitive and specific screening tool for detecting common fetal aneuploidies. The aim of this study was to compare the rates of "atypical" single nucleotide polymorphism (SNP)-based NIPS results and subsequent pregnancy outcomes between Arab American and non-Arab American patients. We conducted a retrospective cohort study of pregnant Arab and non-Arab American patients who had SNP-based NIPS performed between September 2018 and January 2021 at an urban health system in Michigan. The rate of "atypical" results and other perinatal outcomes were compared between groups using descriptive statistics. "Atypical" results due to multifetal gestations, either undisclosed or unknown at time of ordering, were excluded. Five thousand eight hundred and seventy-three patients underwent SNP-based NIPS: 771 (13.1%) were identified as Arab American, 5102 (86.9%) were non-Arab American, and 49 (0.8%) patients received "atypical" results. Arab patients represented only 13.1% of patients screened (771/5873) but had a significantly higher rate of "atypical" results than non-Arab American patients (17/771 [2.2%] vs. 32/5102 [0.6%]; p < 0.001). Of the 17 Arab patients with "atypical" results, 9 (52.9%) were in known consanguineous relationships. No major congenital anomalies or chromosomal aberrations were identified for any patients who had "atypical" results, and no significant differences in other perinatal outcomes were observed between Arab and non-Arab American patients. A better understanding of the association between consanguinity and "atypical" SNP-based NIPS results would aid in appropriate test selection and interpretation and may help physicians and genetic counselors provide better perinatal counseling and follow-up care for patients in consanguineous relationships.

Neonatal Outcomes for Women at Risk of Preterm Delivery Given Half Dose Versus Full Dose of Antenatal Betamethasone: A Randomized, Multicentre, Double-blind, Placebo-Controlled, Non-inferiority Trial

ABSTRACT Antenatal corticosteroid administration to women at risk of preterm delivery is recommended worldwide because of their effectiveness in preventing respiratory distress syndrome, as well as in reducing the rate of necrotizing enterocolitis, intraventricular hemorrhage, and neonatal death in preterm infants. However, the current usual dosage regimen of 2 injections of 12 mg of betamethasone (BMZ) 24 hours apart originated from preclinical experiments in the 1960s and has not been investigated in randomized clinical trials. These authors therefore designed a randomized trial for investigating whether a half dose of BMZ would be as effective as the standard full-dose regimen. The primary outcome was the need for exogenous surfactant within the first 48 hours of life. Secondary outcomes included the incidence and severity of respiratory disease, such as respiratory distress syndrome. This randomized, multicenter, double-blind placebo-controlled, noninferiority study was conducted in 37 perinatal centers in France. Pregnant women 18 years or older with singleton pregnancies at risk of delivery preterm (and already treated with dose 1 of antenatal BMZ before 32 weeks' gestation) were eligible to participate. Exclusion criteria included multiple pregnancy, having previously received full-dose antenatal corticosteroids, 4 cm or greater cervical dilation, cervical length of 20 mm or greater, fetal major malformations or chromosomal aberrations, or being unable to communicate fluently in French. Upon hospital admission after the first BMZ injection, women were provided information about the trial, and those who agreed were randomly assigned in a 1:1 ratio to receive either a saline placebo or the second intramuscular BMZ dose. A total of 3196 women participated, with 1597 in the half-dose group and 1599 in the full-dose group. In all, 30 fetuses were stillborn, 16 neonates were lost to follow-up, and 9 neonates died before evaluation, leaving 1567 neonates in the half-dose group and 1574 neonates in the full-dose group. In the intention-to-treat analysis, the primary outcome occurred in 313 (20%) of neonates in the half-dose group and 276 (17.5%) in the full-dose group (risk difference, 2.4%; 95% confidence interval [CI], 0.3–5.2); thus, noninferiority was not shown. The per-protocol analysis also did not show noninferiority (risk difference, 2.2%; 95% CI, −0.6 to 5.1). There were no differences in the rates of neonatal death, grade 3 to 4 intraventricular hemorrhage, stage ≥2 necrotizing enterocolitis, severe retinopathy of prematurity, or bronchopulmonary dysplasia. Between the 2 groups, all other secondary prematurity-associated outcomes were also similar. Strengths of the study included its large sample size and low loss-to-follow-up rate (0.5%), as well as consistent perinatal management approach through the Groupe de Recherche en Obstetrique et Gynecologie. However, the study also had limitations, such as the potential for the primary outcome to be considered a short-term surrogate only partially reflecting the overall severity of neonatal illness. The rate of the primary outcome observed in the study was also 2.5% lower than anticipated, which the authors attribute to the numbers of deliveries that occurred before 32 weeks' gestation and beyond 7 days following treatment. The number of births occurring within this window of time may limit the ability to demonstrate true differences between the 2 groups, meaning that the authors could not rule out the possibility of half-doses being inferior given higher preterm birth rates. In addition, long-term evaluation of the infants involved in the study via a follow-up evaluation at 5 years of age will be critical to determining the overall effect of BMZ dose reduction on neurological development. The authors concluded that this trial indicated the half-dose antenatal BMZ regimen did not meet criterion for noninferiority when compared with the currently recommended regimen in singleton pregnancies for preventing respiratory distress syndrome that requires exogenous intratracheal surfactant.

Effluent from local palm oil mill refinery in Nigeria is excessively oily and potentially genotoxic

In spite of the fact that the informal, local mills in countries like Nigeria constitute a major portion of the palm oil refinery, adequate attention has not been paid to the quality of the palm oil mill effluent (POME) they generate. In this study, therefore, the physicochemical properties and genotoxic potential of POME generated by a local mill in Nigeria were investigated using the methods prescribed by the American Public Health Association and Allium cepa chromosomal assay, respectively. In addition to the presence of trace/toxic metals (Zn, Pb, Cd, Cr, Cu) and high biochemical oxygen demand, POME contained a very high oil and grease concentration of 10,500 mg L-1 as against the maximum limit of 25 mg L-1 prescribed in the Environmental Management Guideline for the Palm Oil Industry. Mitotic activities in A. cepa roots exposed to POME showed that the number of dividing cells and percentage mitotic index generally decreased with increasing POME concentrations. The major chromosomal aberrations induced by POME were sticky, C-mitosis, bridged anaphase, vagrant, and attached chromosomes. However, no chromosomal aberrations were observed in onion roots exposed to water (control). These results indicate that the local refinery from where the POME was obtained is inefficient at recovering oil from palm fibre. The effluent generated by the mill is also a potential pollutant capable of inducing genotoxic and other adverse effects. These results may be typical of many more local palm oil refineries who use mainly manual methods to extract oil from oil palm fruits.

Abstract 2260: Evaluating a rare t(X;14)(q28;q32) translocation reveals MTCP1 as a driving factor in chronic lymphocytic leukemia

Abstract Chronic lymphocytic leukemia (CLL) is the most prevalent adult leukemia in Western countries and is spelled by substantial genetic and clinical heterogeneity. During CLL transformation, loss or gain of genetic material appears to be a key determinant of disease phenotype and clinical outcome, with major chromosome aberrations observed in up to 80% of patients. Alternatively, balanced translocations, specifically those resulting in constitutive over-expression of various proto-oncogenes under the immunoglobulin heavy chain locus (IGH; 14q32), occur far less frequently. Despite their infrequence, molecular profiling of these rare rearrangements have revealed broad importance of un-recognized genes critical to the pathogenesis of CLL. Employing this strategy, we identified a young CLL patient with a previously undescribed t(X;14)(q28;q32) translocation, co-localization of the mature T cell proliferation 1 (MTCP1; Xq28) coding region with the IGH locus, triggering overexpression of MTCP1 in the CLL cells. Translocations involving MTCP1 are a driving factor in T-prolymphocytic leukemia; however, a role for MTCP1 in CLL has not been described. Inspired by this observation, we screened &amp;gt;1700 suspected CLL cases and evaluated gene expression data for further evidence of MCTP1 aberrations. This query identified seven additional Xq28 rearrangements, revealed MTCP1 mRNA was globally over-expressed in CLL cells compared to normal B-cells, and increased MTCP1 mRNA expression portends a poor response to chemoimmunotherapy. To establish a role for MTCP1 as an oncogene in B cell malignancies, we generated a mouse model with B cell-specific MTCP1 overexpression (Eµ-MTCP1). Longitudinal evaluation revealed a majority of Eµ-MTCP1 mice developed a lethal hematologic malignancy between 5-12 months of age, highlighted by the progressive emergence of clonally related CLL-like B lymphocytes (CD19+/CD5+ B cells) in the blood and accumulating in the spleen and lymph nodes. To support the use of the newly generated Eµ-MTCP1 mouse as a tool for pre-clinical evaluation of CLL therapeutics, we demonstrate that continuous ibrutinib administration in Eµ-MTCP1 mice was sufficient to delay the onset of the CLL-like disease and significantly prolonged survival. In summary, we report Xq28 translocations as rare genetic abnormalities in CLL, yet being one mechanism by which CLL cells amplify expression of MTCP1 compared to normal B cell subsets. Further, the Eµ-MTCP1 mouse model should be considered as an alternative tool for both biologic assessment of co-expressed genes and pre-clinical evaluation of novel CLL therapeutics. Lastly, relevant to all cancer types, successful application of a strategy pursuing the functional consequence of genes involved in rare translocations contributed to the understanding of this disease and identified a novel target for future therapeutic consideration. Citation Format: Janek S. Walker, Zachary A. Hing, Steven Sher, James Cronin, Katie Williams, Bonnie Harrington, Jordan N. Skinner, Casey B. Cempre, Charles T. Gregory, Max Yano, Larry P. Beaver, Brandi R. Walker, Jadwiga M. Labanowska, Nyla A. Heerema, Krzysztof Mrozek, Jennifer A. Woyach, Amy S. Ruppert, Amy Lehman, Hatice Gulcin Ozer, Vincenzo Coppola, John C. Byrd, James S. Blachly, Rosa Lapalombella. Evaluating a rare t(X;14)(q28;q32) translocation reveals MTCP1 as a driving factor in chronic lymphocytic leukemia [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2260.

368P - Genomic analysis of malaysian breast cancers unravel molecular differences from Caucasian breast cancers

BackgroundBreast cancer is the leading cause of cancer death among women, and incidence in Asia is increasing in large part because of changes in reproductive and lifestyle factors. Several appreciable differences exist between breast cancers in Asian and Caucasian women. For example, Asians have a younger median age of incidence and correspondingly, a higher prevalence of hereditary factors, and together, these suggest that they may be crucial differences at the molecular level. MethodsWe performed whole exome sequencing (WES) on 576 Malaysian breast cancers (at median coverage 75X) and their matched normal blood (40X) to detect single nucleotide variations (SNVs) and small insertions and deletions (indels). We also performed shallow whole genome sequencing (sWGS) to detect major chromosomal aberrations, and transcriptomic sequencing (RNA-seq) to measure gene expression. ResultsWe captured known copy number changes, together with major breast cancer genes and their phenotypes, for example high frequency of SNVs in hotspot regions in PIK3CA and indels in GATA3. Interestingly, Malaysian breast cancer show higher prevalence of Her2+ molecular subtypes and TP53 mutations, as well as higher immune scores compared with Caucasian breast cancer cases, consistent with previous findings in other smaller Asian datasets. ConclusionsOur report of the hitherto largest dataset of genomic profiling of Asian breast cancers show the molecular differences between Asian and Caucasian breast cancers and point to potential differences in therapy and outcome. Legal entity responsible for the studyCancer Research Malaysia. FundingCancer Research Malaysia, Medical Research Council Newton Ungku Omar Fund, Scientex Foundation, Estee Lauder Breast Cancer Campaign, Yayasan Sime Darby, Yayasan Petronas. DisclosureAll authors have declared no conflicts of interest.

Physicochemical Quality and Genotoxic Potential of Wastewater Generated by Canteen Complex

Canteens generate high volumes of wastewater that should constantly be subjected to physicochemical and genotoxicity screening. In this study, the wastewater generated by a canteen complex was screened for physicochemical properties and genotoxic potential using standard procedures and Allium cepa chromosome assay. Results showed that the wastewater had total suspended solids, total dissolved solids, and total hardness concentrations of 120.70 mg/l, 554.50 mg/l, and 500.00 mg/l, respectively. The chloride concentration of the wastewater (7873.60 mg/l) was much higher than the recommended limit of 250 mg/l. The wastewater inhibited root growth in A. cepa at 0.1%, 1%, 10%, 25%, 50%, and 100% concentrations but promoted root growth at 2% and 5% concentrations. The wastewater was highly mitodepressive, with mitotic inhibition generally increasing with rising concentrations. The major chromosomal aberrations observed in A. cepa exposed to different concentrations of canteen wastewater were vagrant, sticky, and bridged chromosomes. No chromosomal aberration was observed in onion roots exposed to water (control). The differences in total chromosomal aberrations across wastewater concentrations were not statistically significant (P &gt; 0.05). In view of these results, the practice of discharging untreated canteen wastewater into drainage canals may not be environmentally sustainable.

Genetic variation associated with chromosomal aberration frequency: A genome-wide association study.

Chromosomal aberrations (CAs) in human peripheral blood lymphocytes (PBL) measured with the conventional cytogenetic assay have been used for human biomonitoring of genotoxic exposure for decades. CA frequency in peripheral blood is a marker of cancer susceptibility. Previous studies have shown associations between genetic variants in metabolic pathway, DNA repair and major mitotic checkpoint genes and CAs. We conducted a genome-wide association study on 576 individuals from the Czech Republic and Slovakia followed by a replication in two different sample sets of 482 (replication 1) and 1288 (replication 2) samples. To have a broad look at the genetic susceptibility associated with CA frequency, the sample sets composed of individuals either differentially exposed to smoking, occupational/environmental hazards, or they were untreated cancer patients. Phenotypes were divided into chromosome- and chromatid-type aberrations (CSAs and CTAs, respectively) and total chromosomal aberrations (CAtot). The arbitrary cutoff point between individuals with high and low CA frequency was 2% for CAtot and 1% for CSA and CTA. The data were analyzed using age, sex, occupation/cancer and smoking history as covariates. Altogether 11 loci reached the P-value of 10-5 in the GWAS. Replication 1 supported the association of rs1383997 (8q13.3) and rs2824215 (21q21.1) in CAtot and rs983889 (5p15.1) in CTA analysis. These loci were found to be associated with genes involved in mitosis, response to environmental and chemical factors and genes involved in syndromes linked to chromosomal abnormalities. Identification of new genetic variants for the frequency of CAs offers prediction tools for cancer risk in future. Environ. Mol. Mutagen. 60:17-28, 2019. © 2018 Wiley Periodicals, Inc.

OP22.09: Does NIPS change the spectrum of detected major chromosomal aberrations (CHA)? Population‐based data

Prior to 2012, detection of major CHA had been based mainly on combined first trimester or integrated screening and ultrasound examinations. High-risk cases had been offered invasive diagnostics and karyotyping. The implementation of non-invasive prenatal screening (NIPS) might lead to a decrease of invasive tests causing lower accidental detection of other autosomal aberrations than trisomies 21,18,13 in the population. We tested the hypothesis, that the ratio of prenatally diagnosed CHA has been changed since the gradual implementation of NIPS after 2012. We used data from the National Registry of Congenital Anomalies including the individually collected data on prenatally diagnosed cases. We have analysed the prenatally diagnosed cases of all CHA (ICD-10 codes: Q90-Q99) during the 2012-2015 time period. The numbers were evaluated for the group of main autosomal trisomies and “other” CHA separately. The time trends were statistically evaluated using GraphPad InStat statistical software. The number of invasive procedures, amniocenteses and chorion biopsies decreased from 11407 in the year 2012 to 7680 in 2015. A total of 575 cases of CHA were identified in 2012, of which 329 cases (57.5%) were trisomies 21, 18 and 13. In 2015, 577 cases of all CHA were identified, with major trisomies contributing 374 cases (64.8%). Thus there was no significant change in the proportion of group of major autosomal trisomies, compared to the “other” CHA (p>0.05). During the study period the total number of prenatally diagnosed CHA was 2369; major trisomies contributed 1462 cases (76.3%). We were not able to confirm, that the implementation of NIPS with consequent decrease of invasive procedures would lead to less recognition of other unbalanced CHA, that could also have severe phenotypical features for the fetus and/or infant. The overall ratio of prenatally diagnosed trisomies 21,13,18 and “other” autosomal aberrations remained unchanged. Supported by the Ministry of Health of the Czech Republic RVO VFN64165 and 17-29622A.

Combined comparative genomic hybridization and single-nucleotide polymorphism array detects cryptic chromosomal lesions in both myelodysplastic syndromes and cytopenias of undetermined significance

AbstractThe diagnosis of myelodysplastic syndrome (MDS) can be challenging, and may be facilitated by correlation with cytogenetic testing. Microarray analysis using comparative genomic hybridization and/or single-nucleotide polymorphism array can detect chromosomal abnormalities not seen by standard metaphase cytogenetics. We examined the ability of combined comparative genomic hybridization and single-nucleotide polymorphism analysis (hereafter referred to as ‘combined array') to detect changes among 83 patients with unexplained cytopenias undergoing pathologic evaluation for MDS and compared results with 18 normal bone marrow controls. Thirty-seven patients (45%) were diagnosed with MDS, 12 patients (14%) were demonstrated to have ‘indeterminate dyspoiesis' (insufficient for classification of MDS), 27 (33%) were essentially normal, and 7 patients (8%) had alternative pathologic diagnoses. Twenty-one MDS patients (57% of diagnoses) had effectively normal metaphase cytogenetics, but combined array showed that 5 of these (13% of MDS patients) harbored major cryptic chromosomal aberrations. Furthermore, nearly half of patients with ‘indeterminate dyspoiesis' and 1 with normal morphology had clonal cytopenia(s) of undetermined significance by combined array analysis. Cryptic array findings among MDS patients and those with clonal cytopenias(s) included large-scale copy-neutral loss of heterozygosity (up to 118 Mb) and genomic deletion of loci implicated in MDS pathogenesis (eg, TET2 (4q22) and NUP98 (11p15)). By comparison, in MDS patients with abnormal metaphase cytogenetics, microarray mostly recapitulated findings seen by routine karyotype. Combined array analysis has considerable diagnostic yield in detecting cryptic chromosomal aberrations in MDS and in demonstrating aberrant clonal hematopoiesis in cytopenic patients with indeterminate morphologic dysplasia.

Genotoxic potential of Cynanchum sarcomedium Meve &amp; Liede coupled with its modulatory action on oxidative-stress–mediatedgenotoxicity by hydrogen peroxide

The present study evaluated the genotoxic effect of aqueous extract of C. sarcomedium Meve and Liede and its protective role in chromosomal aberrations induced by 2% hydrogen peroxide (H2O2) on Allium cepa root meristem. Our results showed that the C. sarcomedium extract induced major chromosomal aberrations, viz., nuclear lesions, chromosome breaks, chromosome laggards, vagrance, stickiness, and bridges. These are irreversible in nature and indicate that the plant is a potent clastogen. Total chromosome aberrations were more frequent with increase in exposure time and concentrations of plant extract. For antigenotoxic screening, three different modes of treatment were used: pretreatment, posttreatment, and simultaneous treatment. The significant induction of a wide spectrum of chromosomal aberrations and the reduction in mitotic index effected by the aqueous extract reinforced the genotoxic potential of the plant. The modulatory effect of plant extract was expressed in terms of inhibition percentage and had an excellent effect in reversing chromosomal aberrations induced by H2O2. Dose-dependent results suggest the genotoxic and antigenotoxic effect of C. sarcomedium on onion root tips. These findings offer a useful basis for further investigations on C. sarcomedium, a shrub with a broad array of prospective therapeutic activities.

Major Route Additional Chromosomal Aberrations (ACA) Precede Increase of Blasts in CML: An Analysis of the German CML-Studies III and IIIA

▪During the course of chronic myeloid leukemia (CML) progression to blast crisis (BC) is thought to be caused by genetic instability such as cytogenetic aberrations in addition to the translocation t(9;22)(q34;q11). We have shown previously that major route ACA indicate an unfavorable outcome (Fabarius et al., Blood 2011). We now investigate whether there is a correlation in time between appearance of major route ACA and increase in blast count.Methods: Cytogenetic data and blast count in the peripheral blood were available from 1,290 CML patients recruited to the German CML-studies III (621 patients) and IIIa (669 patients) from January 1995 to January 2004. Treatments were interferon-alpha-based or related allogeneic stem cell transplantation (HSCT). Presence of ACA and major route ACA was considered as a time-dependent covariate. Multivariate proportional hazards models were estimated taking Euro CML score, study III vs. IIIa and stem cell transplantability into account. Cumulative incidences of blast increases were calculated starting at the date of the first ACA or major route ACA, respectively, regarding death as a competing risk. Patients were censored at the date of HSCT with an unrelated donor.Results: 1,287 patients were evaluable with median observation times of 13 and 12 years and a 10-year survival of 48% and 61% in CML studies III and IIIa, respectively. 258 patients progressed to BC with a cumulative 10-year incidence of 20%. 195 patients displayed ACA during the course of disease. 45 patients (15.7%) showed ACA already at diagnosis. 44 patients showed unbalanced minor route, 29 balanced minor route aberrations, 23 -Y. 109 patients showed major route aberrations including 10 with other prior ACA.In a multivariate analysis on 1,257 patients, patients with ACA had a hazard ratio (HR) for a blast increase of between 2.0-2.2 (p<0.001) for blast increases to ≥1%, ≥5%, ≥10%, ≥15%, ≥ 20% and ≥30% compared with patients without ACA (Table). When the same model was performed for major route ACA only at any time during disease, HRs of 2.2-2.7 (p<0.001) were found. For ACA without major route ACA HRs were 1.6-2.1 (p<0.001). In the multivariate analyses of major route ACA vs. no major route ACA a blast increase of 1-5% after diagnosis of major route ACA seems already indicative of progression.5 years after the diagnosis of any ACA the cumulative incidence for a blast increase was 30% (95%- confidence interval (CI): 23-38%), of a major route ACA 40% (95%- CI: 28-49%). The 6-year probability of death without blast increase was 10%. 14 additional patients received an unrelated transplant of which 6 died.We conclude that ACA, particularly major route ACA, precede an increase of blasts. Major route ACA have to be considered as a prognostic indicator for disease progression at any time.Table 1Blast increase toHR (univariate): ACA vs. no ACAHR(multivariate)*: ACA vs. no ACAHR (univariate): major route ACA vs. no major route ACAHR (multivariate)*: major route ACA vs. no major route ACA≥30%2.4092.1392.6462.203≥20%2.4132.1442.6562.211≥15%2.4152.1612.8682.426≥10%2.4162.1602.7992.357≥5%2.2862.0472.7192.278≥1%2.2091.9993.1712.684*adjusted to Euro-Score, study (III vs. IIIa) and transplantability DisclosuresSaussele:ARIAD: Honoraria; BMS: Honoraria, Other: Travel grant, Research Funding; Pfizer: Honoraria, Other: Travel grant; Novartis Pharma: Honoraria, Other: Travel grant, Research Funding. Haferlach:MLL Munich Leukemia Laboratory: Employment, Equity Ownership. Scheid:Janssen-Cilag: Honoraria, Membership on an entity's Board of Directors or advisory committees; Celgene: Honoraria, Membership on an entity's Board of Directors or advisory committees. Baerlocher:Geron Corporation: Research Funding; Novartis: Research Funding. Schnittger:MLL Munich Leukemia Laboratory: Employment, Equity Ownership. Müller:BMS: Consultancy, Honoraria, Research Funding; Ariad: Consultancy, Honoraria, Research Funding; Novartis: Consultancy, Honoraria, Research Funding. Hochhaus:ARIAD: Honoraria, Research Funding; Pfizer: Honoraria, Research Funding; Bristol-Myers Squibb: Honoraria, Research Funding; Novartis: Honoraria, Research Funding. Pfirrmann:BMS: Consultancy, Honoraria; Novartis Pharma: Consultancy, Honoraria. Baccarani:Bristol-Myers Squibb: Honoraria, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; NOVARTIS: Honoraria, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; ARIAD Pharmaceuticals, Inc.: Honoraria, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; PFIZER: Honoraria, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau. Hehlmann:BMS: Consultancy; Novartis Pharma: Research Funding.

Specific genomic and transcriptomic aberrations in tumors induced by partial hepatectomy of a chronically inflamed murine liver.

Resection of hepatocellular carcinoma (HCC) tumors by partial hepatectomy (PHx) is associated with promoting hepatocarcinogenesis. We have previously reported that PHx promotes hepatocarcinogenesis in the Mdr2-knockout (Mdr2-KO) mouse, a model for inflammation-mediated HCC. Now, to explore the molecular mechanisms underlying the tumor-promoting effect of PHx, we compared genomic and transcriptomic profiles of HCC tumors developing in the Mdr2-KO mice either spontaneously or following PHx. PHx accelerated HCC development in these mice by four months. PHx-induced tumors had major chromosomal aberrations: all were amplifications affecting multiple chromosomes. Most of these amplifications were located near the acrocentric centromeres of murine chromosomes. Four different chromosomal regions were amplified each in at least three tumors. The human orthologs of these common amplified regions are known to be amplified in HCC. All tumors of untreated mice had chromosomal aberrations, including both deletions and amplifications. Amplifications in spontaneous tumors affected fewer chromosomes and were not located preferentially at the chromosomal edges. Comparison of gene expression profiles revealed a significantly enriched expression of oncogenes, chromosomal instability markers and E2F1 targets in the post-PHx compared to spontaneous tumors. Both tumor groups shared the same frequent amplification at chromosome 18. Here, we revealed that one of the regulatory genes encoded by this amplified region, Crem, was over-expressed in the nuclei of murine and human HCC cells in vivo, and that it stimulated proliferation of human HCC cells in vitro. Our results demonstrate that PHx of a chronically inflamed liver directed tumor development to a discrete pathway characterized by amplification of specific chromosomal regions and expression of specific tumor-promoting genes. Crem is a new candidate HCC oncogene frequently amplified in this model and frequently over-expressed in human HCC.

Identification of functional cooperative mutations of SETD2 in human acute leukemia

Acute leukemia characterized by chromosomal rearrangements requires additional molecular disruptions to develop into full-blown malignancy, yet the cooperative mechanisms remain elusive. Using whole-genome sequencing of a pair of monozygotic twins discordant for MLL (also called KMT2A) gene-rearranged leukemia, we identified a transforming MLL-NRIP3 fusion gene and biallelic mutations in SETD2 (encoding a histone H3K36 methyltransferase). Moreover, loss-of-function point mutations in SETD2 were recurrent (6.2%) in 241 patients with acute leukemia and were associated with multiple major chromosomal aberrations. We observed a global loss of H3K36 trimethylation (H3K36me3) in leukemic blasts with mutations in SETD2. In the presence of a genetic lesion, downregulation of SETD2 contributed to both initiation and progression during leukemia development by promoting the self-renewal potential of leukemia stem cells. Therefore, our study provides compelling evidence for SETD2 as a new tumor suppressor. Disruption of the SETD2-H3K36me3 pathway is a distinct epigenetic mechanism for leukemia development.

Molecular screening of xq27.3 chromosomal region and cytogenetic abnormalities of fragile-x syndrome in mental retardation peoples of tamil nadu, south india

Objective Fragile X syndrome is the most common inherited form of X-linked mental retardation. The aim of this study was to screen and identify fragile X syndrome-affected individuals in Tamil Nadu (South India) using DNA-based molecular analysis and chromosomal abnormalities. Materials and Methods Genomic DNA extracted from 47 (29 males and 18 females) individuals with mental retardation, including 47 controls, was analyzed using polymerase chain reaction, DNA gel electrophoresis and sequence analysis. Individuals with mutation were determined according to the CGG repeat size in the FMR1gene on chromosome Xq27.3. At same set of mentally retarded individuals, including controls 3ml of blood were collected in Lithium heparin tubes. The blood samples were incubated in 72 hours under culture conditions and harvested for Gbanding technique and results were guaranteed by SKY techniques. Results The sequence analysis shows that the CGG repeat was found in fragile x site which expressed Fragile X syndrome highly when compared to healthy controls. The major Chromosomal aberrations (CA) like deletion, translocation and mosaic were also found. However, when compare with experimental subjects, the controls exhibited low levels of CA (P Conclusion Results of this investigation suggest that, deregulation of the FMR1 gene might have responsible for fragile X syndrome of mental retardation, and CA among the individuals. Further investigation of this study helps to predicts the causes of this syndrome in molecular level (RNA based FMR1gene – Protein).

NOTCH3 is a prognostic factor that promotes glioma cell proliferation, migration and invasion via activation of CCND1 and EGFR.

Using a GWA analysis of a comprehensive glioma specimen population, we identified whole gain of chromosome 19 as one of the major chromosomal aberrations that correlates to patients’ outcomes. Our analysis of significant loci revealed for the first time NOTCH3 as one of the most significant amplification. NOTCH3 amplification is associated with worse outcome compared to tumors with non-amplified locus. NOTCH receptors (NOTCH1-4) are key positive regulators of cell-cell interactions, angiogenesis, cell adhesion and stem cell niche development which have been shown to play critical roles in several human cancers. Our objective is to determine the molecular roles of NOTCH3 in glioma pathogenesis and aggressiveness. Here we show for the first time that NOTCH3 plays a major role in glioma cell proliferation, cell migration, invasion and apoptosis. Therefore, our study uncovers the prognostic value and the oncogenic function of NOTCH3 in gliomagenesis and supports NOTCH3 as a promising target of therapy in high grade glioma. Our studies allowed the identification of a subset of population that may benefit from GSI- or anti-NOTCH3- based therapies. This may lead to the design of novel strategies to improve therapeutic outcome of patients with glioma by establishing medical and scientific basis for personalized chemotherapies.

Establishment and characterization of clear cell renal cell carcinoma cell lines with different metastatic potential from Chinese patients

sBackgroundClear cell renal cell carcinoma (ccRCC) cell lines with distinct metastatic potential are essential to study the mechanism of ccRCC metastasis. However, none of them originated from Chinese.MethodsPrimary cell cultures were performed using a primary tumor of a 49-year-old male ccRCC patient and a metastatic tumor of a 62-year-old male patient who had received nephrectomy to excise primary ccRCC 10 years ago. Cell growth, microstructure, cytogenetics, cytometry, expression of metastasis-associated molecules, tumorigenesis and metastasis were subsequently characterized.ResultsTwo successive cell lines named NRCC from the primary ccRCC and MRCC from the metastatic ccRCC were established, respectively. Compared to NRCC, MRCC exhibited stronger anchorage-independent growth and invasion potentials and contained more glycogen granules in the cytoplasm. Gains of chromosomes and some translocations were the major chromosomal aberrations in both cell strains. CD24 expression was more frequent in MRCC than in NRCC and the same was true for CD56. The transcriptional levels of TNFα, IL-6, VEGF, HIF2α, MMP2, and RhoC were significantly higher in MRCC than in NRCC. Cytosolic IκBα protein was more degraded in MRCC than in NRCC following TNFα treatment. Both cell lines had strong tumorigenicity in athymic nude mice. However, MRCC had strong potential in generating metastasis to lung and hemorrhagic ascites than NRCC following orthotopic transplantations.ConclusionsCancer cells isolated from metastatic ccRCC have more malignant and metastatic potential than those from the primary tumor from the patients who shared the similar race background. Establishment of MRCC and NRCC may provide suitable models with which to investigate molecular mechanisms of ccRCC metastasis.

CYTOGENETIC ANALYSIS OF PROSTATE CARCINOMA PATIENTS

Background and objectives: Prostate carcinoma is the second leading cause of cancer death in men above age of 60 years worldwide. More than 80% of the cancers concomitantly arise with Benign prostate hyperplasia. Prostate carcinoma is associated with various lower urinary tract symptoms, which affect their day to day life. The aim of the present investigation was to find out the major chromosomal aberrations present in Prostate carcinoma patients and to make a comparison with other study. Methods: In present study 05 cases of prostate carcinoma were taken on the basis of clinical diagnosis from the Gujarat Cancer Research Institute (GCRI), Ahmedabad, Gujarat, India, during the period of March 2011 to October 2011. The patients were analyzed for chromosomal aberrations using cultured peripheral blood lymphocytes with their pre-informed written consent. Results: In the present study, all 5 cases were &gt; 60 years of age. 3(60%) cases of prostate carcinoma have elevated PSA level (&gt;4 ng/ml). In adenocarcinoma of prostate gain, deletion, and translocation of 7q22-q31 is common. In present study, Karyotype analysis reveal 01 prostate carcinoma patient had 46,XY,del(7)(q31). In present study 2 cases shows evidence of metastasis. Conclusion: Prostate carcinoma is found in &gt; 60 years of age. Chromosome 1, 2, 3, 6, 7, 9, 16, 22, X and many more chromosomes were affected in prostate carcinoma patients, Common chromosomal aberration is 46,XY,del(7)(q31). Patients with prostate carcinoma have elevated PSA level (&gt;4 ng/ml). Prostate carcinoma associated with metastasis have poor prognosis.

Evaluation Of Etiologic Causes Of Recurrent Pregnancy Loss

SUMMARY Objective: The aim of the present study is to evaluate etiologic factors in patients with recurrent pregnancy loss (RPL). Material and methods: The records of patients admitted to Obstetrics& Gynecology Clinic of Dicle University Medical Faculty Hospital, from 2008 to 2011 were evaluated. Of the 114 patients who all tests were applied for diagnosis in our hospital were enrolled to study. For etiologic evaluation; Parental chromosome analysis, maternal antiphospholipid antibodies, hysterosalpingography for mullerian anomalies, thrombophilic disorders (factor V leiden mutation, prothrombin gene mutation) and endocrine (diabetes mellitus, thyroid hormone disorder) tests applied to patients. Results: The mean age and mean number of abortus were 29.7±6.6, 3.2±1.3 respectively. Fifty (43.9%) patients had at least one etiologic factor where as 64 (56.1%) were idiopathic. Major chromosomal aberrations were detected in four (%3.5) couples as inversion of the ninth chromosome. The frequency for other pathologies; mullerian anomaly:14 (%12.3), factor V leiden mutation:7 (%6.1), prothrombin gene mutation:6 (%5.3) and antiphospholipid antibodies in 10 (%8.8) patients. Conclusion: The prevalance for major chromosomal aberrations, mullerian anomalies, thrombopylic disorders and other pathologic conditions which evaluated in the study were found similar to reported previously. Polymorphism of 9qh+ (heterochromatic centromeric regions) was seen in 39 (%34.2) parents. Further studies are needed to understand that, if this result related to the genetic polymorpisym of the study population? or the pathology associated with RPL?.

Neonatal outcome of 724 children born after ICSI using non-ejaculated sperm

Safety concerns have been expressed regarding the use of immature non-ejaculated spermatozoa for ICSI. Therefore, adverse health outcomes, birth parameters, major anomaly rates and chromosomal aberrations in children born after ICSI using testicular and epididymal sperm were investigated. Questionnaire data and results of physical examinations of 530 children born after ICSI with testicular sperm and of 194 children born after ICSI with epididymal sperm were compared with data on 2516 ICSI children born using ejaculated sperm. Birth parameters, stillborn rates, prematurity rates and rates of low birthweight and very low birthweight were comparable between the non-ejaculated and the ejaculated sperm groups. The perinatal death rate was higher for twins but not for singletons in the non-ejaculated sperm group in comparison to the control cohort of children born using ejaculated sperm. A non-significant increase in major anomalies was reported in the non-ejaculated sperm group in comparison to the ejaculated sperm group. No more anomalies were observed in pre- and post-natal karyotypes from viable pregnancies established using non-ejaculated sperm versus ejaculated sperm. Overall neonatal health in terms of birth parameters, major anomalies and chromosomal aberrations in our large cohort of children born by the use of non-ejaculated sperm seems reassuring in comparison to the outcome of children born after the use of ejaculated sperm.

Apport de l’analyse par hybridation comparative génomique dans le mélanome uvéal : étude de dix cas énucléés

To detect major chromosomal aberrations from enucleated uveal melanoma and relate them to hepatic metastasis and survival. Ten uveal melanomas enucleated between 2005 and 2008 in the Lyon Croix-Rousse Hospital were retrospectively analyzed using a 19000-clone comparative genomic hybridization microarray. The most frequent imbalances were the loss of chromosome 3 (8/10), gain of the 8q arm (7/10) or the entire chromosome 8 (2/10), and gain of the 6p arm (2/10). Most metastatic tumors (6/7) and all cases of death (5/5) concerned melanoma with monosomy 3 and gain of the 8q arm. Genome-wide array comparative genomic hybridization is a reliable tool for identifying uveal melanoma genomic imbalances. Gains of the 8q arm with monosomy 3 are frequent and are strongly associated with poor outcome. Gains of the 6p arm are rare and have a better prognosis. There is a mutually exclusive relationship between monosomy 3 and chromosome 6 abnormalities in our study. These results confirm previously published reports.