The role of microRNA-binding site polymorphisms in DNA repair genes as risk factors for bladder cancer and breast cancer and their impact on radiotherapy outcomes

MicroRNAs (miRNAs) are involved in post-transcriptional regulation of gene expression through binding to messenger RNAs (mRNA) thereby promoting mRNA degradation or altered translation. A single-nucleotide polymorphism (SNP) located within a miRNA-binding site could thus alter mRNA translation and influence cancer risk and treatment response. The common SNPs located within the 3′-untranslated regions of 20 DNA repair genes were analysed for putative miRNA-binding sites using bioinformatics algorithms, calculating the difference in Gibbs free binding energy (ΔΔG) for each wild-type versus variant allele. Seven SNPs were selected to be genotyped in germ line DNAs both from a bladder cancer case–control series (752 cases and 704 controls) and 202 muscle-invasive bladder cancer radiotherapy cases. The PARP-1 SNP rs8679 was also genotyped in a breast cancer case–control series (257 cases and 512 controls). Without adjustment for multiple testing, multivariate analysis demonstrated an association with increased bladder cancer risk with PARP1 rs8679 (Ptrend = 0.05) while variant homozygotes of PARP1 rs8679 were also noted to have an increased breast cancer risk (P = 0.03). In the radiotherapy cases, carriers of the RAD51 rs7180135 minor allele had improved cancer-specific survival (hazard ratio 0.52, 95% confidence interval 0.31–0.87, P = 0.01). This is the first report of associations between DNA repair gene miRNA-binding site SNPs with bladder and breast cancer risk and radiotherapy outcomes. If validated, these findings may give further insight into the biology of bladder carcinogenesis, allow testing of the RAD51 SNP as a potential predictive biomarker and also reveal potential targets for new cancer treatments

Functional assays to determine the significance of two common XPC 3'UTR variants found in bladder cancer patients

<p>Abstract</p> <p>Background</p> <p><it>XPC </it>is involved in the nucleotide excision repair of DNA damaged by carcinogens known to cause bladder cancer. Individuals homozygous for the variant allele of <it>XPC </it>c.1496C > T (p.Ala499Val) were shown in a large pooled analysis to have an increased bladder cancer risk, and we found two 3'UTR variants, *611T > A and c.*618A > G, to be in strong linkage disequilibrium with c.1496T. Here we determined if these two 3'UTR variants can affect mRNA stability and assessed the impact of all three variants on mRNA and protein expression.</p> <p>Methods</p> <p><it>In vitro </it>mRNA stability assays were performed and mRNA and protein expression measured both in plasmid-based assays and in lymphocytes and lymphoblastoid cell lines from bladder and breast cancer patients.</p> <p>Results</p> <p>The two 3'UTR variants were associated with reduced protein and mRNA expression in plasmid-based assays, suggesting an effect on mRNA stability and/or transcription/translation. A near-significant reduction in XPC protein expression (p = 0.058) was detected in lymphoblastoid cell lines homozygous for these alleles but no differences in mRNA stability in these lines was found or in mRNA or protein levels in lymphocytes heterozygous for these alleles.</p> <p>Conclusion</p> <p>The two 3'UTR variants may be the variants underlying the association of c.1496C > T and bladder cancer risk acting via a mechanism modulating protein expression.</p

The impact of cyclin-dependent kinase 5 depletion on poly(ADP-ribose) polymerase activity and responses to radiation

Cyclin-dependent kinase 5 (Cdk5) has been identified as a determinant of sensitivity to poly(ADP-ribose) polymerase (PARP) inhibitors. Here, the consequences of its depletion on cell survival, PARP activity, the recruitment of base excision repair (BER) proteins to DNA damage sites, and overall DNA single-strand break (SSB) repair were investigated using isogenic HeLa stably depleted (KD) and Control cell lines. Synthetic lethality achieved by disrupting PARP activity in Cdk5-deficient cells was confirmed, and the Cdk5KD cells were also found to be sensitive to the killing effects of ionizing radiation (IR) but not methyl methanesulfonate or neocarzinostatin. The recruitment profiles of GFP-PARP-1 and XRCC1-YFP to sites of micro-irradiated Cdk5KD cells were slower and reached lower maximum values, while the profile of GFP-PCNA recruitment was faster and attained higher maximum values compared to Control cells. Higher basal, IR, and hydrogen peroxide-induced polymer levels were observed in Cdk5KD compared to Control cells. Recruitment of GFP-PARP-1 in which serines 782, 785, and 786, potential Cdk5 phosphorylation targets, were mutated to alanines in micro-irradiated Control cells was also reduced. We hypothesize that Cdk5-dependent PARP-1 phosphorylation on one or more of these serines results in an attenuation of its ribosylating activity facilitating persistence at DNA damage sites. Despite these deficiencies, Cdk5KD cells are able to effectively repair SSBs probably via the long patch BER pathway, suggesting that the enhanced radiation sensitivity of Cdk5KD cells is due to a role of Cdk5 in other pathways or the altered polymer levels

Système d'aide au choix et aux réglages du fauteuil roulant manuel

Le fauteuil roulant manuel (FRM) est une aide technique réputée efficace pour augmenter la mobilité de personnes de déficiences très diverses, dans des situations et pour des activités variées. Les catalogues et les sites internet décrivent des gammes de FRM très différentes pour répondre à la variété des besoins réels des utilisateurs. Le FRM est un dispositif mécanique de conception rustique tout en étant un objet complexe car composé de plusieurs centaines de pièces. Il comporte couramment des dizaines de réglages et d'options. Dans ce contexte, les professionnels médicaux doivent préconiser l'usage du FRM, prescrire un type de FRM et aussi préciser les réglages et options, qui conviendront " au mieux " à leur patient. Les distributeurs doivent ensuite trouver chez leurs fournisseurs le ou les FRM qui correspond(ent) à ces prescriptions. En langage scientifique, cette démarche de prescription d'un FRM s'appelle un problème de choix (décision) multicritères. La solution retenue sera forcément un compromis, acceptable (ou satisfaisant). L'objectif de ce projet était de fournir des connaissances, une démarche et des logiciels d'" aide à la décision " à tous ceux qui doivent prescrire ou choisir un FRM. Les connaissances utilisées sont issues d'enquêtes et d'expériences réalisées dans les domaines de la sociologie, de la médecine de rééducation et de la biomécanique. Les résultats obtenus sont déjà ou seront bientôt disponibles dans des articles scientifiques, un livre est également en cours de rédaction et sera publié chez Sauramps, le logiciel sera libre d'accès

Upper limb joint dynamics during manual wheelchair propulsion

Background: Inverse dynamic methods have been widely used to estimate joint loads during manual wheelchair propulsion. However, the interpretation of 3D net joint moments and powers is not always straightforward. It has been suggested to use joint coordinate systems (expression of joint moment on anatomical axes) and the 3D angle between joint moment and angular velocity vectors (propulsion, resistance or stabilization joint configuration) for a better understanding of joint dynamics. Methods: Nine spinal cord injured subjects equipped with reflective markers propelled in a wheelchair with an instrumented wheel. Inverse dynamic results were interpreted using joint coordinate systems, 3D joint power and the 3D angle between the joint moment and joint angular velocity vectors at the three upper limb joints. The 3D angle was used to determine if the joints were predominantly driven (angle close to 0 or 180 degrees) or stabilized (angle close to 90). Findings: The wrist and elbow joints are mainly in a stabilization configuration (angle close to 90) with a combination of extension and ulnar deviation moments and an adduction moment respectively. The shoulder is in a propulsion configuration, but close to stabilization (angle hardly below 60) with a combination of flexion and internal rotation moments. Interpretation: Stabilization configuration at the joints could partly explain the low mechanical efficiency of manual wheelchair propulsion and could give insight about injury risk at the wrist, elbow and shoulder joints

G-quadruplex structures in TP53 intron 3: role in alternative splicing and in production of p53 mRNA isoforms

The tumor suppressor gene TP53, encoding p53, is expressed as several transcripts. The fully spliced p53 (FSp53) transcript encodes the canonical p53 protein. The alternatively spliced p53I2 transcript retains intron 2 and encodes D40p53 (or DNp53), an isoform lacking first 39 N-terminal residues corresponding to the main transactivation domain. We demonstrate the formation of G-quadruplex structures (G4) in a GC-rich region of intron 3 that modulates the splicing of intron 2. First, we show the formation of G4 in synthetic RNAs encompassing intron 3 sequences by ultraviolet melting, thermal difference spectra and circular dichroism spectroscopy. These observations are confirmed by detection of G4-induced reverse transcriptase elongation stops in synthetic RNA of intron 3. In this region, p53 pre-messenger RNA (mRNA) contains a succession of short exons (exons 2 and 3) and introns (introns 2 and 4) covering a total of 333 bp. Site-directed mutagenesis of Gtracts putatively involved in G4 formation decreased by $30% the excision of intron 2 in a green fluorescent protein-reporter splicing assay. Moreover, treatment of lymphoblastoid cells with 360A, a synthetic ligand that binds to single-strand G4 structures, increases the formation of FSp53 mRNA and decreases p53I2 mRNA expression. These results indicate that G4 structures in intron 3 regulate the splicing of intron 2, leading to differential expression of transcripts encoding distinct p53 isoforms