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ИННОВАЦИОННЫЕ ТЕХНОЛОГИИ В ПЕДИАТРИИ И ДЕТСКОЙ ХИРУРГИИ
otide and SNP arrays. In the prenatal case we found mosaicism for an sSMC(4), sSMC(6), sSMC(9), sSMC(14) and sSMC(22),while the postnatal case had an sSMC(4), sSMC(8) and sSMC(11). SNP markers indicated that sSMC(4) resulted from a maternal meiosis II error in the prenatal case and SNP-marker-segregation was consistent with maternal meiosis I error of sSMC(8) in the postnatal case. In the latter, a boy with developmental/psychomo-tor delay, autism, hyperactivity, speech delay and hypotonia, the sSMC(8) was present at the highest frequency in blood and the only mitotically stable sSMC in cultured lymphoblastoid B-cell lines. By comparison with other patients with corresponding duplication a minimal region of overlap was identified, with CHRNB3 and CHRNA6 as dosage-sensitive candidate genes for the phenotype. These genes encode subunits of nicotinic acetylcholine receptors (nAChRs). We propose that overproduction of these sub-units leads to perturbed component stoichiometries with dominant negative effects on the function of nAChRs, as it was shown by others in vitro. With the limitation that in each case only one sSMC could be studied, our findings demonstrate that different meiotic errors lead to multiple sSMCs. Age-related aneuploidy in female meiosis can be a reason here and predivision-sister-chromatid-separation during meiosis I or II, or both seem to generates multiple sSMCs. Part of this research was supported by the Else Kroner-Fresenius-Stiftung (2011_A42).
GENE EXPRESSION PROFILING OF MULTIDRUG RESISTANT CEM/ADR-5000 AND SENSITIVE CCRF-CEM LEUKEMIA CELLS TO SHED LIGHT ON DRUG RESISTANCE MECHANISMS OF TUMORS
Kadioglu O.1, Cao J.1, Kosyakova N.2, Mrasek K.2, Liehr T.2, Efferth T.1
'Department of Pharmaceutical Biology, Institute of Pharmacy and Biochemistry, Johannes Gutenberg University, Mainz, Germany
Jena University Hospital, Friedrich Schiller University, Institute of Human Genetics, Jena, Germany
In order to evaluate differential gene expression profiles, chromosomal alterations and aberrations, multidrug resistant CEM/ADR5000 and sensitive CCRF-CEM leukemia cells were analyzed by RNA sequencing, multicolor fluorescence in situ hybridization (mFISH) and array-comparative genomic hybridization (array-CGH). The gene expression profiles of those leukemia cell lines have not been described well so far and in this study we identified differential gene expression profile of multidrug-resistant (CEM/ADR5000) and sensitive (CCRF-CEM) leukemia cell lines via RNA-sequencing (Illumina HiSeq 2000). Deregulated genes were identified by using the reads per kilobase of exon model per million mapped reads (RPKM) values and they were subjected to downstream pathway and network analysis using Ingenuity Software. Many ATP-binding cassette transporters (ABC) transporters were observed to be deregulated in CEM/ADR5000 cells and deeper analysis showed that pathways and networks linked with multidrug resistance and carcinogenesis were influenced in CEM/ADR5000 cells.
Chromosomal aberrations, translocations, amplifications and deletions were detected by mFISH and array-CGH analyses. The later results were used to validate the RNA-sequencing results. In addition to deletion at chromosome 8, translocation between chromosome 8 and 9 and trisomy at chromosome 20 observed in CCRF-CEM cell line, translocations between chromosomes 3 and 10, 10 and 16, 7 and 18, 9 and 22 and 5,18 and 21, and loss of one X-chromosome were observed in the CEM/ADR5000 cell line. Aberrations were detected within 3q27.1 in CEM/ADR5000 causing slight down-regulation in expression of ABCC5, ABCF3, and within Xq28 causing slight down-regulation in expression of ABCD1. ABCB1, being the critical protein playing role in multidrug resistance, was observed in RNA-seq analyses to be 400-fold up-regulated as a result of an amplification in 7q21.12 observed in array-CGH analyses. This study showed that genomic rearrangements changed gene expression in CEM/ADR5000. Several genes involved in rearrangements were linked to multidrug resistance and carcinogenesis characterizing CEM/ADR5000 cells.
X-LINKED INTELLECTUAL DISABILITY-RELATED GENES DISRUPTED BY BALANCED X-AUTOSOME TRANSLOCATIONS
Moyses-Oliveira M.1, Guilherme R.S.14, Meloni V.A.1, Di B.A.1, de Melo C.B.2, Bragagnolo S.1, Moretti-Ferreira D.3, Kosyakova N.4, Liehr T.4, Carvalheira G.M.1, Melaragno M. I.1
!Genetics Division, Department of Morphology and Genetics, Universidade Federal de Sao Paulo, Sao Paulo, Brazil
2Psychobiology Department,
Universidade Federal de Sao Paulo, Sao Paulo, Brazil 3Departament of Genetics, Instituto de Biociencias de Botucatu, Universidade Estadual de Sao Paulo, Sao Paulo, Brazil
Jena University Hospital, Friedrich Schiller University, Institute of Human Genetics, Jena, Germany
Detailed molecular characterization of chromosomal rearrangements involving X-chromosome has been a key strategy in identifying X-linked intellectual disability-causing genes. We fine-mapped the breakpoints in four women with balanced X-autosome translocations and variable pheno-types in order to investigate the corresponding genetic contribution to intellectual disability. Three patients presented with cognitive impairment, confirming the association between the disrupted genes (TSPAN7-MRX58, KIAA2022-MRX98 and IL1RAPL1 -MRX21/34) and intellectual disability. For TSPAN7 and KIAA202 gene expression analysis showed absence of functional copies in the patient. In the third patient gene expression experiments suggested a fusion transcript ZNF611-IL1RAPL1 under the control of the ZNF611 promoterwith a ZNF611 gene disrupted at the autosomal breakpoint. The X-chromosomal breakpoint in the fourth patient, a woman with normal intellectual abilities, revealed disruption of the ZDHHC15 gene (MRX91). The expression assays did not detect ZDHHC15 gene expression in the patient, thus questioning the involvement of ZDH-
РОССИЙСКИЙ ВЕСТНИКПЕРИНАТОЛОГИИ И ПЕДИАТРИИ, 4, 2015
