CC BY
16
Раздел 2 Педиатрия
HC15 gene in intellectual impairment. Overall, revealing the disruption of an X-linked intellectual disability-related gene in patients with balanced X-autosome translocation is a useful tool for the better characterization of critical genes for neurodevelopment. This approach might be a valuable resource when applied not only in patients with cognitive impairment, but also in subjects with normal cognition.
A NEW IGH@ GENE REARRANGEMENT ASSOCIATED WITH CDKN2A/B DELETION IN A YOUNG ADULT B-CELL ACUTE LYMPHOBLASTIC LEUKEMIA (BALL)
Othman M.A.K.1, Grygalewicz B.2, Pienkowska-Grela B.3, Ejduk A.3, Rincic M.1-4, Melo J.5-6, Carreira I. M.5-6, Meyer B.7, Marzena W.8, Liehr T.1 'Jena University Hospital, Friedrich Schiller University, Institute of Human Genetics, Jena, Germany 2Cytogenetic Laboratory, Maria Sklodowska-Curie Memorial Cancer Centre and Institute, Warsaw, Poland 3Department of Hematology, Institute of Hematology and Transfusion Medicine, Warsaw, Poland "Croatian Institute of Brain Research, Zagreb, Croatia 5Laboratory of Cytogenetics and Genomics, Faculty of Medicine, University of Coimbra, Coimbra, Portugal 6CIMAGO, Centro de Investigagao em Meio Ambiente, Geneticae Oncobiologia 7ZytoVision GmbH, Bremerhaven, Germany 8Department of Haematology and Bone Marrow Transplantation, Holycross Cancer Center, Kielce, Poland
Acquired copy number changes are common in acute leukemia. They are reported as recurrent amplification or deletion and may be indicative for involvement of oncogenes or tumor suppressor genes in the acquired disease and can serve as potential biomarkers for prognosis or even as a target for molecular therapy. Here, we report a gain of copy numbers of 14ql3 to 14q32 leading to an IGH@ locus splitting in a young adult female, present as a yet unreported rearrangement in B-cell acute lymphoblastic leukemia (B-ALL). Low resolution banding cytogenetics at the time of diagnosis revealed a normal karyotype. However, retrospective application of fluorescence in situ hybridization-(FISH-) banding, locus specific FISH-probes, as well as multiplex ligation-dependent probe amplification and high resolution array-comparative genomic hybridization revealed previously cryptic aberrations. Overall a karyotype 46,XX,del(9)(p21.3p21.3),der(14)(pter->q32.33::q32.33->q13::q32.33->qter) was determined. The patient was treated according to PALG 5-ALL7-3 protocol and achieved complete remission. These findings indicate that a favorable prognosis is linked to these aberrations under the mentioned treatment. Supported in parts by the DAAD.
HIGH RATES OF SUBMICROSCOPIC ABERRATIONS IN KARYOTYPICALLY NORMAL ACUTE LYM-PHOBLASTIC LEUKEMIA
Othman M.A.K.1, Melo J.B.23, Carreira I.M.2'3, Rincic M.14, Glaser A.1, Grygalewicz B.5, Gruhn B.6, Wilhelm K.16, Rittscher K.1, Meyer B.7, Macedo S.M.L.8~9, de Jesus Marques S.T.10, Liehr T.1
Jena University Hospital, Friedrich Schiller University, Institute of Human Genetics, Jena, Germany 2Laboratory of Cytogenetics and Genomics, Faculty of Medicine, University of Coimbra, Coimbra, Portugal 3CIMAGO, Centro de Investigagao em Meio Ambiente, Geneticae Oncobiologia, Coimbra, Portugal 4Croatian Institute of Brain Research, Zagreb, Croatia 5Cytogenetic Laboratory, Maria Sklodowska-Curie Memorial Cancer Centre and Institute, Warsaw, Poland •Department of Pediatrics (Oncology and Hematology), Jena University Hospital, Friedrich Schiller University, Jena, Germany
7ZytoVision GmbH, Bremerhaven, Germany Cytogenetics Department, Bone Marrow Transplantation Unit, National Cancer Institute, Rio de Janeiro, RJ, Brazil
9Post Graduation Program in Oncology, National Cancer Institute (INCA), Rio de Janeiro, Brazil 10Pediatric Oncohematology Center, Hospital Oswaldo Cruz/ Pos Graduation Course of the Faculty of Medical Sciences, University of Pernambuco, Recife, PE, Brazil
Acute lymphoblastic leukemia (ALL) is not a single uniform disease. It consists of several subgroups with different cytogenetic and molecular genetic aberrations, clinical presentations and outcomes. Banding cytogenetics plays a pivotal role in the detection of recurrent chromosomal rearrangements and is the starting point of genetic analysis in ALL, still. Nowadays, molecular (cyto)genetic tools provide substantially to identify previously non-detectable, so-called cryptic chromosomal aberrations in ALL. However, ALL according to banding cytogenetics with normal karyotype — in short cytogenetically normal ALL (CN-ALL) — represent up to ~50% of all new diagnosed ALL cases. The overall goal of this study was to identify and characterize the rate of cryptic alterations in CN-ALL and to rule out if one single routine approach may be sufficient to detect most of the cryptic alterations present. 61 ALL patients with CN-ALL were introduced in this study. All of them underwent high resolution fluorescence in situ hybridization (FISH) analysis. Also DNA could be extracted from 34 ALL samples. These DNA-samples were studied using a commercially available MLPA (multiplex ligation-dependent probe amplification) probe set directed against 37 loci in hematological malignancies and/or array-comparative genomic hybridization (aCGH). Chromosomal aberrations were detected in 21 of 61 samples (~34%) applying FISH approaches: structural abnormalities were present in 15 cases and even numerical ones were identified in 6 cases. Applying molecular approaches copy number alterations (CNAs) were detected in 27/34 samples. Overall, 126 CNAs were identified and only 34 of them were detectable by MLPA (~27%). Loss of CNs was identified in ~80% while gain of CNs was present in ~20% of the 126 CNAs. A maximum of 13 aberrations was detected per case; however, only one aberration per case was found in 8 of all in detail studied 34 cases. Of special interest among the detected CNAs are the following new findings: del(15)(q26.1q26.1) including CHD2 gene was found in 20% of the studied ALL cases,
РОССИЙСКИЙ ВЕСТНИКПЕРИНАТОЛОГИИ И ПЕДИАТРИИ, 4, 2015
ИННОВАЦИОННЫЕ ТЕХНОЛОГИИ В ПЕДИАТРИИ И ДЕТСКОЙ ХИРУРГИИ
dup(18)(q21.2q21.2) with the DCC gene was present in 9% of the cases, and the CDK6 gene in 7q21.2 was deleted in 12% of the here in detail studied ALL cases. In conclusion, high resolution molecular cytogenetic tools and molecular approaches like MLPA and aCGH need to be combined in a cost-efficient way, to identify disease and progression causing alterations in ALL, as majority of them are cryptic in banding cytogenetic analyses. This work was supported in parts by DAAD (fellowship to MAKO and PROBRAL 57054562 to TL) and CAPES (419/14 to MLMS).
INTEGRATED GENE MAPPING AND SYNTENY STUDIES GIVE INSIGHTS INTO THE EVOLUTION OF A SEXUAL PROTO-CHROMOSOME IN SOLEA SENEGALENSIS: TESTING INNOVATIVE MOLECULAR CYTOGENETIC APPROACHES FOR BASIC AND DIAGNOSTIC RESEARCH
Portela-Bens S.1, Merlo M.A.1, Rodriguez M.E.1, Cross I.1, Manchado M.2, Kosyakova N.3, Liehr T.3, Rebordinos L.1
'Área de Genética, Facultad de Ciencias del Mar y Ambientales, Universidad de Cádiz, Cádiz, Spain 2Centro IFAPA "El Toruño", Cádiz, Spain 3Institut für Humangenetik, Universitätsklinikum Jena, Jena, Germany
The evolution of genes related to sex and reproduction in fish shows high plasticity; and, to date, the sex determination system has not been identified in more than a few species. Sex determination and sexual differentiation are closely related because genetic and environmental factors work together in a hierarchical network to produce either male or female phenotypes. Solea senegalensis has 42 chromosomes in pairs, and an XX/XY chromosome system for sex determination, while related species show the ZZ/ZW system. Eleven candidate genes involved in these processes (dmtrl, dmtr2, dmtr3, sox3, sox6, sox, sox9, lh, cya19a1a, amh, vasa, aqp3, and nanos3) have been studied. Sequencing produced 56 contigs, from which 93 genes could be annotated. By including 9 previous mapped? bacterial artificial chromosomes (BACs) and one containing the fshß gene an integrated map has been produced, in which 15 pairs out of 21 have at least one BAC, important for distinguishing those chromosomes of S. senegalensis being similar in shape and size. The sequenced size of the 21 BACs represents about 10 % of the total size of the genome of Senegalensis. Multicolor fluorescence in situ hybridization (mFISH) showed the following co-locations in the same chromosomes: dmrt1-dmrt2-dmrt3, dmtr4-sox9-thrß, aqp3-sox8, cyp19a1a-fshß, igsf9b-sox3 and lysg-sox6. Synteny studies showed that the co-location of dmrt1-dmrt2-drmt3 in the largest metacentric chromosome of S. senegalensis is coincident with the Z chromosome of Cynoglossus semilaevis — a finding that would make this potentially a proto-sex-ual chromosome. Phylogenetic studies of 11 concatenated candidate genes show the close proximity of S. senegalensis to Danio rerio, in which temperature during early life determines sex. Comparative mapping with other species provides evidence of the preferential association of these candidate genes in particular chromosomes, instead of random
distribution. This suggests a mechanism for the selection of blocks of genes, with important repercussions in other species. This study has been supported by the Spanish Ministerio de Ciencia e Innovación MICINN (AGL 2011-25596) and by the AQUAGENET project (SOE 2/P1/E287) from the INTERREG IVB SUDOE program. SPB received a fellowship from the UCA.
A CRYPTIC THREE-WAY TRANSLOCATION T(10;19;11)(P12.31;Q13.31;Q23.3) WITH A DERIVATIVE Y-CHROMOSOME IN AN INFANT WITH ACUTE MYELOBLASTIC LEUKEMIA (M5B)
Rittscher K.1, Othman M.A.K.1, Vujic D.2-3, ZZecevic Z.3, Durisic M.3, Slavkovic B.3, Meyer B.4, Liehr T.1
'Jena University Hospital, Friedrich Schiller University, Institute of Human Genetics, Jena, Germany 2University of Belgrade Faculty of Medicine, Belgarde, Serbia
3Mother and Child Health Care Institution of Serbia "Dr. Vukan Cupic", Belgrade, Serbia 4ZytoVision GmbH, Bremerhaven, Germany
Acute myeloid leukemia (AML) is a heterogeneous disease characterized by the malignant transformation of hemato-poietic precursors to a pathogenic cell clone. Chromosomal band 11q23 harboring MLL (= mixed lineage leukemia) gene is known to be involved in rearrangements with variety of genes as activating partners MLL in different AML subtypes. Overall, an unfavorable prognosis is associated with MLL abnormalities. Here we investigated an 11-month-old male presenting with hyperleukocytosis being diagnosed with AML subtype FAB-M5b. In banding cytogenetics a der(19)t(19;?)(q13.3;?) and del(Y)(q11.23) were found as sole aberrations. Molecular cytogenetics revealed that the MLL gene was disrupted and even partially lost due to a t(10;19;11)(p12.31;q13.31;q23.3), an MLL/MLLT10 fusion appeared, and the der(Y) was an asymmetric inverted duplication with breakpoints in Yp11.2 and Yq11.23. The patient got hematopoietic stem cell transplantation from his haploidentical mother. Still three months afterwards 15% of blasts were detected in bone marrow and later patient was lost during follow up. The present case highlights the necessity to exclude MLL rearrangements, even when there seems to be no actual hint from banding cytogenetics. Supported in parts by the DAAD.
COMPLEX CHROMOSOMAL REARRANGEMENTS IN PERIPHERAL BLOOD AND DECREASED AMOUNT OF NATURAL KILLER CELLS IN TWO FEMALE GAS STATION ATTENDANTS OF RIO DE JANEIRO
Santiago F.1'2'3, Lima S.1, Pinheiro T.1, Tavares S.R.1A3, Otero U.B.4, Tabalipa M.M.4, Kosyakova N.5, Ornellas M.H.12, Liehr T.5, Alves GM3
1Laboratório de Marcadores Circulantes, Departamento de Patologia e Laboratórios, Faculdade de Ciencias Médicas, Universidade do Estado do Rio de Janeiro, Rio de Janeiro, Brasil 2Pós-graduagao em Ciencias Médicas (PGCM),
РОССИЙСКИЙ ВЕСТНИКПЕРИНАТОЛОГИИ И ПЕДИАТРИИ, 4, 2015
