The dual role of the X-linked FoxP3 gene in human cancers

The FoxP3 (forkhead box P3) gene is an X-linked gene that is submitted to inactivation. It is an essential transcription factor in CD4(+)CD25(+)FoxP3 regulatory T cells, which are therapeutic targets in disseminated cutaneous melanoma. Moreover, FoxP3 is an important tumor suppressor gene in carcinomas and has putative cancer suppressor gene function in cutaneous melanoma as well. Therefore understanding the structure and function of the FoxP3 gene is crucial to gaining insight into the biology of melanoma to better develop immunotherapeutics and future therapeutic strategies

BORIS/CTCFL promotes a switch from a proliferative towards an invasive phenotype in melanoma cells

Melanoma is among the most aggressive cancers due to its tendency to metastasize early. Phenotype switching between a proliferative and an invasive state has been suggested as a critical process for metastasis, though the mechanisms that regulate state transitions are complex and remain poorly understood. Brother of Regulator of Imprinted Sites (BORIS), also known as CCCTC binding factor-Like (CTCFL), is a transcriptional modulator that becomes aberrantly expressed in melanoma. Yet, the role of BORIS in melanoma remains elusive. Here, we show that BORIS is involved in melanoma phenotype switching. Genetic modification of BORIS expression in melanoma cells combined with whole-transcriptome analysis indicated that BORIS expression contributes to an invasion-associated transcriptome. In line with these findings, inducible BORIS overexpression in melanoma cells reduced proliferation and increased migration and invasion, demonstrating that the transcriptional switch is accompanied by a phenotypic switch. Mechanistically, we reveal that BORIS binds near the promoter of transforming growth factor-beta 1 (TFGB1), a well-recognized factor involved in the transition towards an invasive state, which coincided with increased expression of TGFB1. Overall, our study indicates a pro-invasive role for BORIS in melanoma via transcriptional reprogramming

MOLLI T1 mapping versus T2 W-SPAIR at 3T : myocardial area at risk measurements and the influence of microvascular obstruction

Funding Information: This study was supported by a Medical Research Council (UK) grant, as a sub-study of Nitrites in Acute Myocardial Infarction, NCT01388504.Peer reviewe

T₁ mapping for assessment of myocardial injury and microvascular obstruction at one week post myocardial infarction

OBJECTIVES: To compare 3T T1 mapping to conventional T2-weighted (T2W) imaging for delineating myocardial oedema one week after ST-elevation myocardial infarction (STEMI), and to explore the confounding effects of microvascular obstruction (MVO) on each technique.  METHODS: T2W spectral attenuated inversion recovery and native T1 mapping were applied in 10 healthy volunteers and 62 STEMI patients, and late gadolinium enhancement was included for infarct localisation at 1 week and at 6 months post-STEMI. Segmental T1 values and T2W signal intensity ratios were calculated; oedema volumes and salvage indices were determined in patients using image thresholding-a receiver operator characteristic (ROC) derived T1 threshold, and a 2SD T2W threshold; and the results were compared between patients with/without MVO (n=35/27).  RESULTS: Native T1 mapping delineated oedema with significantly better discriminatory power than T2W-as indicated by ROC analysis (area-under-the-curve, AUC=0.89 versus 0.83, p=0.009; and sensitivity/specificity=83/83% versus 73/73%). The optimal ROC threshold derived for T1 mapping was 1241ms, which gave significantly larger oedema volumes than 2SD T2W (p=0.006); with this threshold, patients with and without MVO showed similar oedema volumes, but patients with MVO had significantly poorer salvage indices (p<0.05) than those without. Neither method was significantly affected by MVO, the volume of which was seen to increase exponentially with infarct size.  CONCLUSIONS: Native T1 mapping at 3T can delineate oedema one week post-STEMI, showing larger oedema volumes and better discriminatory power than T2W imaging, and it is suitable for quantitative thresholding. Both techniques are robust against MVO-related magnetic susceptibility

The PREDICTS database: a global database of how local terrestrial biodiversity responds to human impacts

Biodiversity continues to decline in the face of increasing anthropogenic pressures such as habitat destruction, exploitation, pollution and introduction of alien species. Existing global databases of species’ threat status or population time series are dominated by charismatic species. The collation of datasets with broad taxonomic and biogeographic extents, and that support computation of a range of biodiversity indicators, is necessary to enable better understanding of historical declines and to project – and avert – future declines. We describe and assess a new database of more than 1.6 million samples from 78 countries representing over 28,000 species, collated from existing spatial comparisons of local-scale biodiversity exposed to different intensities and types of anthropogenic pressures, from terrestrial sites around the world. The database contains measurements taken in 208 (of 814) ecoregions, 13 (of 14) biomes, 25 (of 35) biodiversity hotspots and 16 (of 17) megadiverse countries. The database contains more than 1% of the total number of all species described, and more than 1% of the described species within many taxonomic groups – including flowering plants, gymnosperms, birds, mammals, reptiles, amphibians, beetles, lepidopterans and hymenopterans. The dataset, which is still being added to, is therefore already considerably larger and more representative than those used by previous quantitative models of biodiversity trends and responses. The database is being assembled as part of the PREDICTS project (Projecting Responses of Ecological Diversity In Changing Terrestrial Systems – www.predicts.org.uk). We make site-level summary data available alongside this article. The full database will be publicly available in 2015

Unraveling the role of the X chromosome in cancer: characterization of FOXP3 Isoforms and PR70 in cutaneous melanoma

Many cancers have a different incidence, behavior and response to treatment in men and women. Traditionally these sex-specific variations have been attributed to differences in habits, occupational exposures and hormones. More recently it has been recognized that some of the dissimilarities can be attributed to genetics. We were particularly struck by the significant increase in the incidence and mortality of melanoma in men. The objective of this project was to characterize how the X chromosome contributes to this effect. Global analysis of genomic alterations revealed that a somatic loss of one X chromosome in females was significantly associated with a worse prognosis in melanoma. We further showed that it was the inactive X that was lost in all cases. This led to the hypothesis that an activating mutation of an oncogene, or inactivating mutation of a tumor suppressor gene, on the remaining X chromosome in these patients was contributing to tumor aggressiveness. PR70 and FOXP3 were identified as warranting further investigation as candidate X-linked tumor suppressors in melanoma based on their behaviour in other tumor types. The PPP2R3B gene (Xp22.33) encodes for PR70, the regulatory subunit of a protein phosphatase that plays a critical role in cell cycle progression. In melanoma a strong correlation is demonstrated between low levels of PR70 mRNA expression and poor distant metastasis-free survival by multivariate analyses. In line with this, no or low PR70 protein expression is associated with poor overall survival in three independent sample sets of melanoma. Endogenous and exogenous PR70 expression levels are shown to have an inverse correlation with the rate of melanoma cell proliferation. Exogenous PR70 overexpression decreases tumor take and growth in nude mice compared to shRNA-mediated downregulation that increases it. The FOXP3 gene (Xp23.11) encodes a transcription factor that is important for T-regulatory cell development. Interestingly, FOXP3 was also found to be expressed in the majority of our melanoma cell lines at both the mRNA and protein level. All five of the known isoforms of FOXP3 were shown to localize to the nucleus in melanoma cell lines and demonstrated isoform- and cell line-specific effects on gene transcription. Both PR70 and FOXP3 are associated with a dose-dependent decrease in melanoma cell proliferation with an accumulation of cells in the G1 phase of the cell cycle. These findings are significant because proliferative rate is one of the strongest independent prognostic factors in melanoma.De nombreux cancers ont une incidence, un comportement et une réponse au traitement différents chez les hommes et chez les femmes. Traditionnellement, ces variations spécifiques liées au sexe ont été attribuées à des différences entre les sexes dans les habitudes, l'exposition professionnelle et le statut hormonal. Plus récemment, il a été reconnu que certaines de ces différences peuvent être attribuées à la génétique. Nous avons été particulièrement frappés par l'augmentation significative de l'incidence et de la mortalité du mélanome chez l'homme. L'objectif de ce projet était de caractériser la contribution du chromosome X à cet effet. L'analyse globale des altérations génomiques a révélée qu'une perte somatique d'un chromosome X chez les femmes est associée de manière significative à un pronostic plus défavorable du mélanome. Nous avons en outre montré que c'est le X inactif qui est perdu dans tous les cas. Cela a conduit à l'hypothèse qu'une mutation activatrice d'un oncogène, ou une mutation inactivatrice d'un gène suppresseur de tumeur sur le chromosome X restant chez ces patients contribuent à l'agressivité de la tumeur. PPP2R3B et FOXP3 ont été identifiés comme des gènes justifiant une investigation plus poussée en tant que gènes suppresseurs de tumeurs liés au chromosome X dans le mélanome en raison de leur comportement dans d'autres types de tumeurs. Nous avons démontré que ces gènes sont exprimés dans la majorité de nos lignées cellulaires au niveau de l'ARNm et des protéines. L'expression de PR70 endogène et exogène est significativement corrélée à une diminution de la prolifération. Dans le mélanome humain, il existe une forte corrélation entre les faibles taux d'expression de l'ARNm du PR70 et la faible survie sans métastase à distance par les analyses multivariées. Dans cette optique, une expression nulle ou faible de la protéine PR70 est associée à une faible survie globale dans trois cohortes indépendantes de mélanome. La surexpression de PR70 exogène diminue la prise de tumeur et la croissance chez les souris nues, contrairement à la baisse de l'expression de PR70 induite par shRNA qui l'augmente. PR70 et le FOXP3 sont tous deux associés à une diminution dose-dépendante de la prolifération cellulaire du mélanome avec une accumulation de cellules dans la phase G1 du cycle cellulaire. Ces résultats sont d'importance in vivo car la prolifération est l'un des facteurs pronostiques indépendants les plus forts dans le mélanome

Scoring system avoids Chlamydia trachomatis overscreening in women seeking surgical abortions

International audienceOBJECTIVE: To develop and validate a predictive score to avoid unnecessary screening and prophylactic antibiotic use in abortion clinics by identifying a group of women who are at very low risk for Chlamydia trachomatis (CT) infection. METHODS: This population-based retrospective study includes 1000 women who underwent surgical abortion between January and September 2010. The main outcome measure was the rate of CT infection among women seeking an induced abortion according to sociodemographic and clinical data. The score was developed by using two-thirds of the data set as the derivation sample to identify the strongest predictors of CT. A receiver operating characteristic curve established cutoffs and applied the score to the remaining one-third (validation sample). RESULTS: The rate of CT infection was 6.7%. Three criteria were independently associated with CT: gestation more than 10 weeks (adjusted odds ratio [aOR], 1.96; 95% confidence interval [95% CI], 1.06-3.64), not using contraception (aOR, 2.70; 95% CI, 1.41-5.16), and having 0 or 1 child (aOR, 3.46; 95% CI, 1.34-8.93). The CT score was based on these 3 criteria. The low-risk group was derived from values of the score (probability of CT, 1.3% [95% CI, 0-3.0]). Application of these criteria to the validation data set confirmed the diagnostic accuracy of the score (probability of CT, 0%). Sensitivity was 100% and specificity was 26.9% for the score in the validation data set. When applied to the validation data set, the score avoided 25.4% of CT tests and screened 100% of CT-infected women before surgical abortion. CONCLUSIONS: This easy-to-calculate score may prove useful for avoiding CT test in 25% of patients seeking surgical abortion

Study protocol for a phase II dose evaluation randomized controlled trial of cholecalciferol in critically ill children with vitamin D deficiency (VITdAL-PICU study)

Abstract Background Clinical research has recently demonstrated that vitamin D deficiency (VDD) is highly prevalent in the pediatric intensive care unit (PICU) and associated with worse clinical course. Multiple adult ICU trials have suggested that optimization of vitamin D status through high-dose supplementation may reduce mortality and improve other clinically relevant outcomes; however, there have been no trials of rapid normalization in the PICU setting. The objective of this study is to evaluate the safety and efficacy of an enteral weight-based cholecalciferol loading dose regimen in critically ill children with VDD. Methods/design The VITdAL-PICU pilot study is designed as a multicenter placebo-controlled phase II dose evaluation pilot randomized controlled trial. We aim to randomize 67 VDD critically ill children using a 2:1 randomization schema to receive loading dose enteral cholecalciferol (10,000 IU/kg, maximum of 400,000 IU) or a placebo solution. Participants, caregivers and outcome assessors will be blinded to allocation. Eligibility criteria include ICU patient, aged 37 weeks to 18 years, expected ICU length of stay more than 48 h, anticipated access to bloodwork at 7 days, and VDD (blood total 25 hydroxyvitamin D < 50 nmol/L). The primary objective is to determine whether the dosing protocol normalizes vitamin D status, defined as a blood total 25(OH)D concentration above 75 nmol/L. Secondary objectives include an examination of the safety of the dosing regimen (e.g. hypercalcemia, hypercalciuria, nephrocalcinosis), measures of vitamin D axis function (e.g. calcitriol levels, immune function), and protocol feasibility (eligibility criteria, protocol deviations, blinding). Discussion Despite significant observational literature suggesting VDD to be a modifiable risk factor in the PICU setting, there is no robust clinical trial evidence evaluating the benefits of rapid normalization. This phase II clinical trial will evaluate an innovative weight-based dosing regimen intended to rapidly and safely normalize vitamin D levels in critically ill children. Study findings will be used to inform the design of a multicenter phase III trial evaluating the clinical and economic benefits to rapid normalization. Recruitment for this trial was initiated in January 2016 and is expected to continue until November 30, 2017. Trial registration Clinicaltrials.gov NCT0245276

Additional file 3: of Prevention of vitamin D deficiency in children following cardiac surgery: study protocol for a randomized controlled trial

Flow chart of study-related safety measures. (JPEG 166 kb