TY - JOUR A1 - Weis, Eva A1 - Schoen, Holger A1 - Victor, Anja A1 - Spix, Claudia A1 - Ludwig, Marco A1 - Schneider-Raetzke, Brigitte A1 - Kohlschmidt, Nicolai A1 - Bartsch, Oliver A1 - Gerhold-Ay, Aslihan A1 - Boehm, Nils A1 - Grus, Franz A1 - Haaf, Thomas A1 - Galetzka, Danuta T1 - Reduced mRNA and Protein Expression of the Genomic Caretaker RAD9A in Primary Fibroblasts of Individuals with Childhood and Independent Second Cancer JF - PLoS ONE N2 - Background: The etiology of secondary cancer in childhood cancer survivors is largely unclear. Exposure of normal somatic cells to radiation and/or chemotherapy can damage DNA and if not all DNA lesions are properly fixed, the mis-repair may lead to pathological consequences. It is plausible to assume that genetic differences, i.e. in the pathways responsible for cell cycle control and DNA repair, play a critical role in the development of secondary cancer. Methodology/Findings: To identify factors that may influence the susceptibility for second cancer formation, we recruited 20 individuals who survived a childhood malignancy and then developed a second cancer as well as 20 carefully matched control individuals with childhood malignancy but without a second cancer. By antibody microarrays, we screened primary fibroblasts of matched patients for differences in the amount of representative DNA repair-associated proteins. We found constitutively decreased levels of RAD9A and several other DNA repair proteins in two-cancer patients, compared to one-cancer patients. The RAD9A protein level increased in response to DNA damage, however to a lesser extent in the two-cancer patients. Quantification of mRNA expression by real-time RT PCR revealed lower RAD9A mRNA levels in both untreated and 1 Gy gamma-irradiated cells of two-cancer patients. Conclusions/Significance: Collectively, our results support the idea that modulation of RAD9A and other cell cycle arrest and DNA repair proteins contribute to the risk of developing a second malignancy in childhood cancer patients. KW - DNA methylation KW - Malignant neoplasms KW - Genes KW - Instability KW - Stability KW - Susceptibility KW - Checkpoints KW - Repair KW - Damage Y1 - 2011 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-141838 VL - 6 IS - 10 ER - TY - JOUR A1 - Weis, Eva A1 - Schoen, Holger A1 - Victor, Anja A1 - Spix, Claudia A1 - Ludwig, Marco A1 - Schneider-Raetzke, Brigitte A1 - Kohlschmidt, Nicolai A1 - Bartsch, Oliver A1 - Gerhold-Ay, Aslihan A1 - Boehm, Nils A1 - Grus, Franz A1 - Haaf, Thomas A1 - Galetzka, Danuta T1 - Reduced mRNA and Protein Expression of the Genomic Caretaker RAD9A in Primary Fibroblasts of Individuals with Childhood and Independent Second Cancer N2 - Background: The etiology of secondary cancer in childhood cancer survivors is largely unclear. Exposure of normal somatic cells to radiation and/or chemotherapy can damage DNA and if not all DNA lesions are properly fixed, the mis-repair may lead to pathological consequences. It is plausible to assume that genetic differences, i.e. in the pathways responsible for cell cycle control and DNA repair, play a critical role in the development of secondary cancer. Methodology/Findings: To identify factors that may influence the susceptibility for second cancer formation, we recruited 20 individuals who survived a childhood malignancy and then developed a second cancer as well as 20 carefully matched control individuals with childhood malignancy but without a second cancer. By antibody microarrays, we screened primary fibroblasts of matched patients for differences in the amount of representative DNA repair-associated proteins. We found constitutively decreased levels of RAD9A and several other DNA repair proteins in two-cancer patients, compared to onecancer patients. The RAD9A protein level increased in response to DNA damage, however to a lesser extent in the twocancer patients. Quantification of mRNA expression by real-time RT PCR revealed lower RAD9A mRNA levels in both untreated and 1 Gy c-irradiated cells of two-cancer patients. Conclusions/Significance: Collectively, our results support the idea that modulation of RAD9A and other cell cycle arrest and DNA repair proteins contribute to the risk of developing a second malignancy in childhood cancer patients. KW - Medizin Y1 - 2011 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-74777 ER - TY - JOUR A1 - Galetzka, Danuta A1 - Hansmann, Tamara A1 - El Hajj, Nady A1 - Weis, Eva A1 - Irmscher, Benjamin A1 - Ludwig, Marco A1 - Schneider-Rätzke, Brigitte A1 - Kohlschmidt, Nicolai A1 - Beyer, Vera A1 - Bartsch, Oliver A1 - Zechner, Ulrich A1 - Spix, Claudia A1 - Haaf, Thomas T1 - Monozygotic twins discordant for constitutive BRCA1 promoter methylation, childhood cancer and secondary cancer JF - Epigenetics N2 - We describe monozygotic twins discordant for childhood leukemia and secondary thyroid carcinoma. We used bisulfite pyrosequencing to compare the constitutive promoter methylation of BRCA1 and several other tumor suppressor genes in primary fibroblasts. The affected twin displayed an increased BRCA1 methylation (12%), compared with her sister (3%). Subsequent bisulfite plasmid sequencing demonstrated that 13% (6 of 47) BRCA1 alleles were fully methylated in the affected twin, whereas her sister displayed only single CpG errors without functional implications. This between-twin methylation difference was also found in irradiated fibroblasts and untreated saliva cells. The BRCA1 epimutation may have originated by an early somatic event in the affected twin: approximately 25% of her body cells derived from different embryonic cell lineages carry one epigenetically inactivated BRCA1 allele. This epimutation was associated with reduced basal protein levels and a higher induction of BRCA1 after DNA damage. In addition, we performed a genome-wide microarray analysis of both sisters and found several copy number variations, i.e., heterozygous deletion and reduced expression of the RSPO3 gene in the affected twin. This monozygotic twin pair represents an impressive example of epigenetic somatic mosaicism, suggesting a role for constitutive epimutations, maybe along with de novo genetic alterations in recurrent tumor development. KW - BRCA1 KW - childhood cancer KW - DNA Methylation KW - epimutation KW - monozygotic twins KW - secondary cancer KW - somatic mosaicism Y1 - 2012 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-125386 VL - 7 IS - 1 ER - TY - JOUR A1 - Semmler, Anna-Lena A1 - Sacconi, Sabrina A1 - Bach, J. Elisa A1 - Liebe, Claus A1 - Bürmann, Jan A1 - Kley, Rudolf A. A1 - Ferbert, Andreas A1 - Anderheiden, Roland A1 - Van den Bergh, Peter A1 - Martin, Jean-Jacques A1 - De Jonghe, Peter A1 - Neuen-Jacob, Eva A1 - Müller, Oliver A1 - Deschauer, Marcus A1 - Bergmann, Markus A1 - Schröder, J. Michael A1 - Vorgerd, Matthias A1 - Schulz, Jörg B. A1 - Weis, Joachim A1 - Kress, Wolfram A1 - Claeys, Kristl G. T1 - Unusual multisystemic involvement and a novel BAG3 mutation revealed by NGS screening in a large cohort of myofibrillar myopathies JF - Orphanet Journal of Rare Diseases N2 - Background: Myofibrillar myopathies (MFM) are a group of phenotypically and genetically heterogeneous neuromuscular disorders, which are characterized by protein aggregations in muscle fibres and can be associated with multisystemic involvement. Methods: We screened a large cohort of 38 index patients with MFM for mutations in the nine thus far known causative genes using Sanger and next generation sequencing (NGS). We studied the clinical and histopathological characteristics in 38 index patients and five additional relatives (n = 43) and particularly focused on the associated multisystemic symptoms. Results: We identified 14 heterozygous mutations (diagnostic yield of 37%), among them the novel p. Pro209Gln mutation in the BAG3 gene, which was associated with onset in adulthood, a mild phenotype and an axonal sensorimotor polyneuropathy, in the absence of giant axons at the nerve biopsy. We revealed several novel clinical phenotypes and unusual multisystemic presentations with previously described mutations: hearing impairment with a FLNC mutation, dysphonia with a mutation in DES and the first patient with a FLNC mutation presenting respiratory insufficiency as the initial symptom. Moreover, we described for the first time respiratory insufficiency occurring in a patient with the p. Gly154Ser mutation in CRYAB. Interestingly, we detected a polyneuropathy in 28% of the MFM patients, including a BAG3 and a MYOT case, and hearing impairment in 13%, including one patient with a FLNC mutation and two with mutations in the DES gene. In four index patients with a mutation in one of the MFM genes, typical histological findings were only identified at the ultrastructural level (29%). Conclusions: We conclude that extraskeletal symptoms frequently occur in MFM, particularly cardiac and respiratory involvement, polyneuropathy and/or deafness. BAG3 mutations should be considered even in cases with a mild phenotype or an adult onset. We identified a genetic defect in one of the known genes in less than half of the MFM patients, indicating that more causative genes are still to be found. Next generation sequencing techniques should be helpful in achieving this aim. KW - polyneuropathy KW - MFM KW - next generation sequencing KW - bcl-2 associated athanogene protein 3 KW - protein aggregation KW - hearing impairment KW - early respiratory-failure KW - myopathy KW - muscular-dystrophy KW - skeletal myopathy Y1 - 2014 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-115623 SN - 1750-1172 N1 - Additional files are available here: http://www.ojrd.com/content/9/1/121/additional VL - 9 IS - 121 ER -