PMC5771875_01
Representative molecular findings. (A) Array CGH and SNP array analyses for chromosome 14. The upper two panels show array CGH findings. The vertical axis shows log2 signal ratios, and the signals around +0.5, +-0, and -1.0 indicate duplicated, normal, and deleted segments, respectively. A region with >=three consecutive dots that are present in increased or decreased log2 signal ratios is regarded as a copy number variant, and there is no such a region in this patient as well as in a control subject. The lower two panels show the SNP array findings. Most SNPs are present in a homozygous status (+1.0 or -1.0), and a small number of SNPs are apparently present in a heterozygous status (+-0) in this patient, whereas examined SNPs are present in both homozygous and heterozygous conditions in the control subject. (B) Methylation indices (MIs, the frequencies of methylated clones) of the CpG sites within nine DMRs examined by pyrosequencing. The black circles show the MIs of this patient. Gray vertical bars indicate the ranges of MIs (minimum-maximum) in 50 control subjects, and the red bars for the MEG3/DLK1:IG-DMR and MEG3:TSS-DMR represent the range of MIs (minimum-maximum) in 11 KOS patients with upd(14)pat. (C) Microsatellite analysis. Major peaks of paternal origin and minor peaks of maternal origin (red asterisks) are identified in this patient. For D14S258, while the minor peak could be of paternal or maternal origin, overall SNP array data and microsatellite data indicate that the minor peak is of maternal origin (see Note S1). L: leukocytes, and B: buccal cells.
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SD), and her head circumference 55.5 cm (+1.1 SD) 2. She still exhibited somewhat characteristic facial appearance and had large hands and feet and joint contractures (Fig. S1B). Cytoscan HD (Affymetrix, Santa Clara, CA). Cytogenetic studies showed a normal 46,XX karyotype in two lymphocyte metaphase spreads examined, and aCGH and SNP array analyses revealed the absence of pathogenic copy number variant and the presence of apparent but not complete isodisomy for whole chromosome 14 (iso-upd(14)pat) with a small number of heterozygous SNPs (Fig. 1A). Consequently, we showed isolated hypermethylations of the MEG3/DLK1:IG-DMR and MEG3:TSS-DMR, with the methylation indices for the two DMRs being higher than those of 50 control subjects and lower than those of 11 KOS patients with upd(14)pat (Fig. 1B). We further performed microsatellite analysis for 10 loci on chromosome 14, using leukocyte gDNA of the patient and the parents (for primers, see Table S2), demonstrating the presence of cells with maternally inherited chromosome 14 (Fig. 1C and Table S2).
Chest roentgenogram at 9 years of age. AMNR: age-matched normal range in the Japanese children 3.
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We further performed microsatellite analysis for 10 loci on chromosome 14, using leukocyte gDNA of the patient and the parents (for primers, see Table S2), demonstrating the presence of cells with maternally inherited chromosome 14 (Fig. 1C and Table S2). Furthermore, KOS-like thoracic and/or rib abnormalities were not recorded for chest roentgenograms obtained in the early neonatal period (unfortunately, roentgenograms were not preserved), and chest roentgenograms obtained at 9 years of age for a minor chest trauma showed that the CHA was slightly above the normal range and the M/W ratio remained within the normal range (Fig. 2).