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1.
Cancer Med ; 7(4): 1349-1358, 2018 04.
Article in English | MEDLINE | ID: mdl-29522266

ABSTRACT

The prevalence of germ line mutations in non-BRCA1/2 genes associated with hereditary breast cancer (BC) is low, and the role of some of these genes in BC predisposition and pathogenesis is conflicting. In this study, 5589 consecutive BC index patients negative for pathogenic BRCA1/2 mutations and 2189 female controls were screened for germ line mutations in eight cancer predisposition genes (ATM, CDH1, CHEK2, NBN, PALB2, RAD51C, RAD51D, and TP53). All patients met the inclusion criteria of the German Consortium for Hereditary Breast and Ovarian Cancer for germ line testing. The highest mutation prevalence was observed in the CHEK2 gene (2.5%), followed by ATM (1.5%) and PALB2 (1.2%). The mutation prevalence in each of the remaining genes was 0.3% or lower. Using Exome Aggregation Consortium control data, we confirm significant associations of heterozygous germ line mutations with BC for ATM (OR: 3.63, 95%CI: 2.67-4.94), CDH1 (OR: 17.04, 95%CI: 3.54-82), CHEK2 (OR: 2.93, 95%CI: 2.29-3.75), PALB2 (OR: 9.53, 95%CI: 6.25-14.51), and TP53 (OR: 7.30, 95%CI: 1.22-43.68). NBN germ line mutations were not significantly associated with BC risk (OR:1.39, 95%CI: 0.73-2.64). Due to their low mutation prevalence, the RAD51C and RAD51D genes require further investigation. Compared with control datasets, predicted damaging rare missense variants were significantly more prevalent in CHEK2 and TP53 in BC index patients. Compared with the overall sample, only TP53 mutation carriers show a significantly younger age at first BC diagnosis. We demonstrate a significant association of deleterious variants in the CHEK2, PALB2, and TP53 genes with bilateral BC. Both, ATM and CHEK2, were negatively associated with triple-negative breast cancer (TNBC) and estrogen receptor (ER)-negative tumor phenotypes. A particularly high CHEK2 mutation prevalence (5.2%) was observed in patients with human epidermal growth factor receptor 2 (HER2)-positive tumors.


Subject(s)
Biomarkers, Tumor , Genes, BRCA1 , Genes, BRCA2 , Hereditary Breast and Ovarian Cancer Syndrome/diagnosis , Hereditary Breast and Ovarian Cancer Syndrome/genetics , Adult , Aged , Aged, 80 and over , Case-Control Studies , Female , Genetic Association Studies , Genetic Predisposition to Disease , Genetic Testing/methods , Genetic Variation , Hereditary Breast and Ovarian Cancer Syndrome/epidemiology , Humans , Middle Aged , Odds Ratio , Prevalence , Young Adult
2.
Nurs Stand ; 30(50): 30, 2016 Aug 10.
Article in English | MEDLINE | ID: mdl-27507381

ABSTRACT

Key workers 'affordable' housing is shocking treatment.


Subject(s)
Housing/economics , Housing/standards , Nurses/psychology , Humans , United Kingdom
3.
Mol Genet Genomic Med ; 3(5): 467-80, 2015 Sep.
Article in English | MEDLINE | ID: mdl-26436113

ABSTRACT

Seckel syndrome is a heterogeneous, autosomal recessive disorder marked by prenatal proportionate short stature, severe microcephaly, intellectual disability, and characteristic facial features. Here, we describe the novel homozygous splice-site mutations c.383+1G>C and c.4005-9A>G in CDK5RAP2 in two consanguineous families with Seckel syndrome. CDK5RAP2 (CEP215) encodes a centrosomal protein which is known to be essential for centrosomal cohesion and proper spindle formation and has been shown to be causally involved in autosomal recessive primary microcephaly. We establish CDK5RAP2 as a disease-causing gene for Seckel syndrome and show that loss of functional CDK5RAP2 leads to severe defects in mitosis and spindle organization, resulting in cells with abnormal nuclei and centrosomal pattern, which underlines the important role of centrosomal and mitotic proteins in the pathogenesis of the disease. Additionally, we present an intriguing case of possible digenic inheritance in Seckel syndrome: A severely affected child of nonconsanguineous German parents was found to carry heterozygous mutations in CDK5RAP2 and CEP152. This finding points toward a potential additive genetic effect of mutations in CDK5RAP2 and CEP152.

4.
J Clin Invest ; 125(9): 3585-99, 2015 Sep.
Article in English | MEDLINE | ID: mdl-26280580

ABSTRACT

The genetic disorder Kabuki syndrome (KS) is characterized by developmental delay and congenital anomalies. Dominant mutations in the chromatin regulators lysine (K)-specific methyltransferase 2D (KMT2D) (also known as MLL2) and lysine (K)-specific demethylase 6A (KDM6A) underlie the majority of cases. Although the functions of these chromatin-modifying proteins have been studied extensively, the physiological systems regulated by them are largely unknown. Using whole-exome sequencing, we identified a mutation in RAP1A that was converted to homozygosity as the result of uniparental isodisomy (UPD) in a patient with KS and a de novo, dominant mutation in RAP1B in a second individual with a KS-like phenotype. We elucidated a genetic and functional interaction between the respective KS-associated genes and their products in zebrafish models and patient cell lines. Specifically, we determined that dysfunction of known KS genes and the genes identified in this study results in aberrant MEK/ERK signaling as well as disruption of F-actin polymerization and cell intercalation. Moreover, these phenotypes could be rescued in zebrafish models by rebalancing MEK/ERK signaling via administration of small molecule inhibitors of MEK. Taken together, our studies suggest that the KS pathophysiology overlaps with the RASopathies and provide a potential direction for treatment design.


Subject(s)
Abnormalities, Multiple/genetics , Exome , Face/abnormalities , Hematologic Diseases/genetics , MAP Kinase Signaling System/genetics , Monomeric GTP-Binding Proteins/genetics , Mutation , Telomere-Binding Proteins/genetics , Vestibular Diseases/genetics , Zebrafish Proteins/genetics , Abnormalities, Multiple/metabolism , Actins/genetics , Actins/metabolism , Animals , Cattle , Child , Child, Preschool , DNA-Binding Proteins/genetics , DNA-Binding Proteins/metabolism , Disease Models, Animal , Hematologic Diseases/metabolism , Histone Demethylases/genetics , Histone Demethylases/metabolism , Humans , Male , Mice , Monomeric GTP-Binding Proteins/metabolism , Neoplasm Proteins/genetics , Neoplasm Proteins/metabolism , Nuclear Proteins/genetics , Nuclear Proteins/metabolism , Rats , Shelterin Complex , Telomere-Binding Proteins/metabolism , Vestibular Diseases/metabolism , Zebrafish , Zebrafish Proteins/metabolism
5.
Hum Genet ; 133(1): 29-39, 2014 Jan.
Article in English | MEDLINE | ID: mdl-23982343

ABSTRACT

Proteoglycan (PG) synthesis begins with the sequential addition of a "linker chain", made up of four sugar residues, to a specific region of a core protein. Defects in the enzymes catalyzing steps two to four of the linker chain synthesis have been shown to cause autosomal recessive human phenotypes while no mutation has yet been reported in humans for the xylosyltransferases 1 and 2 (XT1 and XT2), the initiating enzymes in the linker chain formation. Here, we present a consanguineous Turkish family with two affected individuals presenting with short stature, distinct facial features, alterations of fat distribution, and moderate intellectual disability. X-rays showed only mild skeletal changes in the form of a short femoral neck, stocky and plump long bones and thickened ribs. Using a combination of whole-exome sequencing (WES), determination of homozygous stretches by WES variants, and classical linkage analysis, we identified the homozygous missense mutation c.C1441T in XYLT1, encoding XT1, within a large homozygous stretch on chromosome 16p13.12-p12.1. The mutation co-segregated with the phenotype in the family, is not found in over 13,000 alleles in the exome variant server and is predicted to change a highly conserved arginine at position 481 (p.R481W) located in the putative catalytical domain. Immunostaining of primary patient fibroblasts showed a loss of predominance of Golgi localization in mutant cells. Moreover, western blot analysis of decorin in cell culture supernatant demonstrated glycosylation differences between patient and control cells. Our data provide evidence that functional alterations of XT1 cause an autosomal recessive short stature syndrome associated with intellectual disability.


Subject(s)
Dwarfism/genetics , Intellectual Disability/genetics , Pentosyltransferases/genetics , Consanguinity , Exome , Female , Genes, Recessive , Genetic Linkage , Homozygote , Humans , Male , Mutation, Missense , Pedigree , Pentosyltransferases/metabolism , Phenotype , Proteoglycans/metabolism , Sequence Analysis, DNA , Syndrome , Turkey , X-Rays , UDP Xylose-Protein Xylosyltransferase
6.
Hum Mol Genet ; 22(25): 5121-35, 2013 Dec 20.
Article in English | MEDLINE | ID: mdl-23906836

ABSTRACT

Chromatin remodeling complexes are known to modify chemical marks on histones or to induce conformational changes in the chromatin in order to regulate transcription. De novo dominant mutations in different members of the SWI/SNF chromatin remodeling complex have recently been described in individuals with Coffin-Siris (CSS) and Nicolaides-Baraitser (NCBRS) syndromes. Using a combination of whole-exome sequencing, NGS-based sequencing of 23 SWI/SNF complex genes, and molecular karyotyping in 46 previously undescribed individuals with CSS and NCBRS, we identified a de novo 1-bp deletion (c.677delG, p.Gly226Glufs*53) and a de novo missense mutation (c.914G>T, p.Cys305Phe) in PHF6 in two individuals diagnosed with CSS. PHF6 interacts with the nucleosome remodeling and deacetylation (NuRD) complex implicating dysfunction of a second chromatin remodeling complex in the pathogenesis of CSS-like phenotypes. Altogether, we identified mutations in 60% of the studied individuals (28/46), located in the genes ARID1A, ARID1B, SMARCB1, SMARCE1, SMARCA2, and PHF6. We show that mutations in ARID1B are the main cause of CSS, accounting for 76% of identified mutations. ARID1B and SMARCB1 mutations were also found in individuals with the initial diagnosis of NCBRS. These individuals apparently belong to a small subset who display an intermediate CSS/NCBRS phenotype. Our proposed genotype-phenotype correlations are important for molecular screening strategies.


Subject(s)
Abnormalities, Multiple/genetics , Chromatin Assembly and Disassembly/genetics , Face/abnormalities , Foot Deformities, Congenital/genetics , Hand Deformities, Congenital/genetics , Hypotrichosis/genetics , Intellectual Disability/genetics , Micrognathism/genetics , Neck/abnormalities , Sequence Deletion/genetics , Abnormalities, Multiple/pathology , Adolescent , Adult , Carrier Proteins/genetics , Child , Child, Preschool , Chromosomal Proteins, Non-Histone/genetics , DNA-Binding Proteins/genetics , Exome/genetics , Face/pathology , Facies , Female , Foot Deformities, Congenital/pathology , Hand Deformities, Congenital/pathology , High-Throughput Nucleotide Sequencing , Humans , Hypotrichosis/pathology , Infant , Infant, Newborn , Intellectual Disability/pathology , Karyotyping , Male , Micrognathism/pathology , Mutation, Missense , Neck/pathology , Repressor Proteins , SMARCB1 Protein , Transcription Factors/genetics
7.
Hum Genet ; 132(11): 1311-20, 2013 Nov.
Article in English | MEDLINE | ID: mdl-23851939

ABSTRACT

Otofaciocervical syndrome (OFCS) is an autosomal recessively inherited disorder characterized by facial dysmorphism, external ear anomalies with preauricular pits and hearing impairment, branchial cysts or fistulas, anomalies of the vertebrae and the shoulder girdle, and mild intellectual disability. In a large consanguineous family with OFCS from Turkey, we performed whole-exome sequencing (WES) of a single pooled DNA sample of four affected individuals. Filtering for variants with a percentage of alternate reads ≥ 90 % and a coverage of at least five reads identified only a single novel homozygous variant, c.497G>T, located in PAX1 that co-segregated with the disease in the family. PAX1 encodes a transcription factor with a critical role in pattern formation during embryogenesis in vertebrates. The mutation is predicted to substitute the glycine at position 166 to valine (p.G166V) within the highly conserved paired-box domain of the PAX1 protein. We performed a dual luciferase reporter assay to examine the transactivation of a regulatory sequence in the Nkx3-2 promoter region, which is a direct target of mouse Pax1 transcriptional regulation. We observed a significantly reduced transactivation in HEK293T cells overexpressing Pax1(G157V) in comparison to Pax1(WT) expressing cells, indicating a reduced DNA-binding affinity of the mutant protein. Taken together, our results show that the strategy of pooling DNA is a powerful, cost-effective application for WES in consanguineous families and establish PAX1 as a new disease-causing gene for OFCS and as part of the EYA-DACH-SIX-PAX network, important in early embryogenesis.


Subject(s)
Branchio-Oto-Renal Syndrome/genetics , Paired Box Transcription Factors/genetics , Amino Acid Sequence , Animals , Female , Gene Expression Regulation, Developmental , HEK293 Cells , Homeodomain Proteins/genetics , Homeodomain Proteins/metabolism , Humans , Male , Mice , Molecular Sequence Data , Mutation , Paired Box Transcription Factors/metabolism , Pedigree , Promoter Regions, Genetic , Sequence Analysis, DNA , Transcription Factors/genetics , Transcription Factors/metabolism , Turkey
8.
Am J Hum Genet ; 90(4): 661-74, 2012 Apr 06.
Article in English | MEDLINE | ID: mdl-22482805

ABSTRACT

Bone morphogenetic protein 1 (BMP1) is an astacin metalloprotease with important cellular functions and diverse substrates, including extracellular-matrix proteins and antagonists of some TGFß superfamily members. Combining whole-exome sequencing and filtering for homozygous stretches of identified variants, we found a homozygous causative BMP1 mutation, c.34G>C, in a consanguineous family affected by increased bone mineral density and multiple recurrent fractures. The mutation is located within the BMP1 signal peptide and leads to impaired secretion and an alteration in posttranslational modification. We also characterize a zebrafish bone mutant harboring lesions in bmp1a, demonstrating conservation of BMP1 function in osteogenesis across species. Genetic, biochemical, and histological analyses of this mutant and a comparison to a second, similar locus reveal that Bmp1a is critically required for mature-collagen generation, downstream of osteoblast maturation, in bone. We thus define the molecular and cellular bases of BMP1-dependent osteogenesis and show the importance of this protein for bone formation and stability.


Subject(s)
Bone Morphogenetic Protein 1/physiology , Osteogenesis/genetics , Osteogenesis/physiology , Animals , Base Sequence , Bone Density Conservation Agents/therapeutic use , Bone Morphogenetic Protein 1/genetics , Bone Morphogenetic Protein 1/metabolism , Bone and Bones/metabolism , Cell Differentiation , Child, Preschool , Collagen/biosynthesis , Diphosphonates/therapeutic use , Exome , Female , Fractures, Bone/drug therapy , Fractures, Bone/prevention & control , Genetic Loci , Heat-Shock Proteins , Humans , Male , Molecular Sequence Data , Mutation , Osteoblasts/drug effects , Osteoblasts/physiology , Osteogenesis/drug effects , Peptide Fragments , Protein Processing, Post-Translational , Zebrafish/genetics , Zebrafish/metabolism
9.
Hum Genet ; 130(6): 715-24, 2011 Dec.
Article in English | MEDLINE | ID: mdl-21607748

ABSTRACT

Kabuki syndrome (KS) is one of the classical, clinically well-known multiple anomalies/mental retardation syndromes, mainly characterized by a very distinctive facial appearance in combination with additional clinical signs such as developmental delay, short stature, persistent fingerpads, and urogenital tract anomalies. In our study, we sequenced all 54 coding exons of the recently identified MLL2 gene in 34 patients with Kabuki syndrome. We identified 18 distinct mutations in 19 patients, 11 of 12 tested de novo. Mutations were located all over the gene and included three nonsense mutations, two splice-site mutations, six small deletions or insertions, and seven missense mutations. We compared frequencies of clinical symptoms in MLL2 mutation carriers versus non-carriers. MLL2 mutation carriers significantly more often presented with short stature and renal anomalies (p = 0.026 and 0.031, respectively), and in addition, MLL2 carriers obviously showed more frequently a typical facial gestalt (17/19) compared with non-carriers (9/15), although this result was not statistically significant (p = 0.1). Mutation-negative patients were subsequently tested for mutations in ten functional candidate genes (e.g. MLL, ASC2, ASH2L, and WDR5), but no convincing causative mutations could be found. Our results indicate that MLL2 is the major gene for Kabuki syndrome with a wide spectrum of de novo mutations and strongly suggest further genetic heterogeneity.


Subject(s)
Abnormalities, Multiple/genetics , Hematologic Diseases/genetics , Mutation , Vestibular Diseases/genetics , DNA Mutational Analysis , DNA-Binding Proteins/genetics , Exons , Face/abnormalities , Female , Genetic Heterogeneity , Heterozygote , Humans , Male , Neoplasm Proteins/genetics , Phenotype , Sequence Analysis, DNA
10.
Nat Genet ; 43(1): 23-6, 2011 Jan.
Article in English | MEDLINE | ID: mdl-21131973

ABSTRACT

Functional impairment of DNA damage response pathways leads to increased genomic instability. Here we describe the centrosomal protein CEP152 as a new regulator of genomic integrity and cellular response to DNA damage. Using homozygosity mapping and exome sequencing, we identified CEP152 mutations in Seckel syndrome and showed that impaired CEP152 function leads to accumulation of genomic defects resulting from replicative stress through enhanced activation of ATM signaling and increased H2AX phosphorylation.


Subject(s)
Cell Cycle Proteins/genetics , Genome, Human , Child , Child, Preschool , DNA Damage , Dwarfism/genetics , Facies , Genomic Instability , Histones/genetics , Humans , Male , Microcephaly/genetics , Mutation , Phosphorylation
11.
Am J Hum Genet ; 86(3): 479-84, 2010 Mar 12.
Article in English | MEDLINE | ID: mdl-20170898

ABSTRACT

We performed genome-wide homozygosity mapping in a large consanguineous family from Morocco and mapped the autosomal-recessive nonsyndromic hearing loss (ARNSHL) in this family to the DFNB79 locus on chromosome 9q34. By sequencing of 62 positional candidate genes of the critical region, we identified a causative homozygous 11 bp deletion, c.42_52del, in the TPRN gene in all seven affected individuals. The deletion is located in exon 1 and results in a frameshift and premature protein truncation (p.Gly15AlafsX150). Interestingly, the deleted sequence is part of a repetitive and CG-rich motive predicted to be prone to structural aberrations during crossover formation. We identified another family with progressive ARNSHL linked to this locus, whose affected members were shown to carry a causative 1 bp deletion (c.1347delG) in exon 1 of TPRN. The function of the encoded protein, taperin, is unknown; yet, partial homology to the actin-caping protein phostensin suggests a role in actin dynamics.


Subject(s)
Hearing Loss/genetics , Mutation , Proteins/genetics , Amino Acid Sequence , Animals , Base Sequence , Chromosome Mapping , Chromosomes, Human, Pair 9/genetics , Cochlea/metabolism , Consanguinity , DNA/genetics , DNA Primers/genetics , Exons , Female , Frameshift Mutation , Gene Expression , Genes, Recessive , Hearing Loss/congenital , Homozygote , Humans , Male , Mice , Morocco , Pedigree , Sequence Deletion
12.
J Neurosci Res ; 85(5): 1057-64, 2007 Apr.
Article in English | MEDLINE | ID: mdl-17335079

ABSTRACT

In the present study, we compare the capacity of two different embryonic stem (ES) cell lines to secrete neurotrophins in response to cerebral tissue extract derived from healthy or injured rat brains. The intrinsic capacity of the embryonic cell lines BAC7 (feeder cell-dependent cultivation) to release brain-derived neurotrophic factor (BDNF) or neurotrophin-3 (NT-3) exceeded the release of these factors by CGR8 cells (feeder cell-free growth) by factors of 10 and 4, respectively. Nerve growth factor (NGF) was secreted only by BAC7 cells. Conditioning of cell lines with cerebral tissue extract derived from healthy or fluid percussion-injured rat brains resulted in a significant time-dependent increase in BDNF release in both cell lines. The increase in BDNF release by BAC7 cells was more pronounced when cells were incubated with brain extract derived from injured brain. However, differences in neurotrophin release associated with the origin of brain extract were at no time statistically significant. Neutrophin-3 and NGF release was inhibited when cell lines were exposed to cerebral tissue extract. The magnitude of the response to cerebral tissue extract was dependent on the intrinsic capacity of the cell lines to release neurotrophins. Our results clearly demonstrate significant variations in the intrinsic capability of different stem cell lines to produce neurotrophic factors. Furthermore, a significant modulation of neurotrophic factor release was observed following conditioning of cell lines with tissue extract derived from rat brains. A significant modulation of neurotrophin release dependent on the source of cerebral tissue extract used was not observed.


Subject(s)
Cerebral Cortex/metabolism , Embryonic Stem Cells/metabolism , Nerve Growth Factors/metabolism , Animals , Brain Chemistry , Brain Tissue Transplantation/methods , Brain-Derived Neurotrophic Factor/metabolism , Cell Differentiation/drug effects , Cell Differentiation/physiology , Cell Extracts/pharmacology , Cell Line , Cerebral Cortex/cytology , Culture Media, Conditioned/pharmacology , Mice , Nerve Regeneration/drug effects , Nerve Regeneration/physiology , Neurotrophin 3/metabolism , Stem Cell Transplantation/methods , Time Factors
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