Integrative analysis of transcriptomics and clinical data uncovers the tumor- suppressive activity of MITF in prostate cancer

The dysregulation of gene expression is an enabling hallmark of cancer. Computational analysis of transcriptomics data from human cancer specimens, complemented with exhaustive clinical annotation, provides an opportunity to identify core regulators of the tumorigenic process. Here we exploit well-annotated clinical datasets of prostate cancer for the discovery of transcriptional regulators relevant to prostate cancer. Following this rationale, we identify Microphthalmia-associated transcription factor (MITF) as a prostate tumor suppressor among a subset of transcription factors. Importantly, we further interrogate transcriptomics and clinical data to refine MITF perturbation-based empirical assays and unveil Crystallin Alpha B (CRYAB) as an unprecedented direct target of the transcription factor that is, at least in part, responsible for its tumor-suppressive activity in prostate cancer. This evidence was supported by the enhanced prognostic potential of a signature based on the concomitant alteration of MITF and CRYAB in prostate cancer patients. In sum, our study provides proof-of-concept evidence of the potential of the bioinformatics screen of publicly available cancer patient databases as discovery platforms, and demonstrates that the MITF-CRYAB axis controls prostate cancer biology

Identification of novel Coxiella burnetii genotypes from Ethiopian ticks

Background: Coxiella burnetii , the etiologic agent of Q fever, is a highly infectious zoonotic bacterium. Genetic information about the strains of this worldwide distributed agent circulating on the African continent is limited. The aim of the present study was the genetic characterization of C. burnetii DNA samples detected in ticks collected from Ethiopian cattle and their comparison with other genotypes found previously in other parts of the world. Methodology/Principal Findings: A total of 296 tick samples were screened by real-time PCR targeting the IS 1111 region of C. burnetii genome and from the 32 positive samples, 8 cases with sufficient C. burnetii DNA load ( Amblyomma cohaerens ,n 5 6; A. variegatum ,n 5 2) were characterized by multispacer sequence typing (MST) and multiple-locus variable-number tandem repeat analysis (MLVA). One novel sequence type (ST), the proposed ST52, was identified by MST. The MLVA-6 discriminated the proposed ST52 into two newly identified MLVA genotypes: type 24 or AH was detected in both Amblyomma species while type 26 or AI was found only in A. cohaerens . Conclusions/Significance: Both the MST and MLVA genotypes of the present work are closely related to previously described genotypes found primarily in cattle samples from different parts of the globe. This finding is congruent with the source hosts of the analyzed Ethiopian ticks, as these were also collected from cattle. The present study provides genotype information of C. burnetii from this seldom studied East-African region as well as further evidence for the presumed host-specific adaptation of this agent

Genomic and functional regulation of TRIB1 contributes to prostate cancer pathogenesis

Prostate cancer is the most frequent malignancy in European men and the second worldwide. One of the major oncogenic events in this disease includes amplification of the transcription factor cMYC. Amplification of this oncogene in chromosome 8q24 occurs concomitantly with the copy number increase in a subset of neighboring genes and regulatory elements, but their contribution to disease pathogenesis is poorly understood. Here we show that TRIB1 is among the most robustly upregulated coding genes within the 8q24 amplicon in prostate cancer. Moreover, we demonstrate that TRIB1 amplification and overexpression are frequent in this tumor type. Importantly, we find that, parallel to its amplification, TRIB1 transcription is controlled by cMYC. Mouse modeling and functional analysis revealed that aberrant TRIB1 expression is causal to prostate cancer pathogenesis. In sum, we provide unprecedented evidence for the regulation and function of TRIB1 in prostate cancer

Genetic manipulation of LKB1 elicits lethal metastatic prostate cancer

Gene dosage is a key defining factor to understand cancer pathogenesis and progression, which requires the development of experimental models that aid better deconstruction of the disease. Here, we model an aggressive form of prostate cancer and show the unconventional association of LKB1 dosage to prostate tumorigenesis. Whereas loss of Lkbl alone in the murine prostate epithelium was inconsequential for tumorigenesis, its combination with an oncogenic insult, illustrated by Pten heterozygosity, elicited lethal metastatic prostate cancer. Despite the low frequency of LKB1 deletion in patients, this event was significantly enriched in lung metastasis. Modeling the role of LKB1 in cellular systems revealed that the residual activity retained in a reported kinase-dead form, LKB1(K781), was sufficient to hamper tumor aggressiveness and metastatic dissemination. Our data suggest that prostate cells can function normally with low activity of LKB1, whereas its complete absence influences prostate cancer pathogenesis and dissemination

Estimation of Coxiella burnetii prevalence in dairy cattle in intensive systems by serological and molecular analyses of bulk-tank milk samples

A large-scale investigation on Coxiella burnetii was carried out in dairy cattle herds from a Q fever-endemic region to evaluate the degree of exposure to C. burnetii and to estimate prevalences. This study included all of the dairy cattle herds from the province of Bizkaia, Northern Spain (n=178). Herds were visited between September 2009 and February 2010, and 100mL of bulk-tank milk (BTM) per farm was collected to be analyzed by ELISA and PCR. Blood samples were also taken from about 15 animals randomly selected from each herd. One hundred nineteen of the 178 studied herds (66.9±6.9%) were positive for the presence of anti-C. burnetii antibodies in BTM. Serum samples from 1,306 cows, 654 heifers, and 502 calves were analyzed by ELISA, and cows showed a statistically significantly higher seroprevalence (12.3±1.8%) than heifers (1.1±0.8%) and calves (0.0±0.0%). Eighty-nine herds (50.0±7.3%) had at least 1 seropositive animal, but within-herd prevalences higher than 20% were only observed in 24 herds (13.5±5.0%). A significant correlation was observed between BTM ELISA sample-to-positive control ratios and within-herd seroprevalence, being higher when considering only cows (R 2=0.21). Animals from herds with negative BTM by ELISA showed a mean seroprevalence of 2.5%, whereas animals from herds with positive BTM samples had a statistically significantly higher seroprevalence (8.9%, F=19.7, degrees of freedom=1). The proportion of herds C. burnetii positive by BTM PCR was 51.7±7.3% (92/178). The widespread distribution of C. burnetii in cattle advocates for the implementation of Q fever control strategies.This work was supported by funding from Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA, Madrid, Spain; RTA 2009-00017-00), European Regional Development Fund (ERDF), and the Department of Agriculture of the Basque Government (Spain). I. Astobiza is the recipient of a predoctoral fellowship from the Department of Agriculture, Fisheries and Food of the Basque Government and A. Piñero is the recipient of a predoctoral fellowship from INIA.Peer Reviewe

Coxiella burnetii shedding and environmental contamination at lambing in two highly naturally-infected dairy sheep flocks after vaccination

Abortion due to Coxiella burnetii was confirmed in the 2007/08 season in two naturally-infected dairy sheep flocks. Proportion of C. burnetii shedders and bacterial loads in vaginal mucus were high among aborted or lambed ewes, as was within-flock seroprevalence. Before the next reproductive season (2008/09) 75% of ewes and 50% of replacement lambs were vaccinated (Coxevac, CEVA Santé Animale) keeping the remaining as untreated controls. Compared with the previous year results when abortion outbreak started, a great reduction in the percentage of abortions, in the number of shedders and in the bacterial burden excreted by the ewes was found in both flocks. However, seroconversion in non-vaccinated yearlings from both flocks and the presence of C. burnetii DNA in bioaerosols taken at sheep premises at lambing indicated that infection was still active. No differences were observed between vaccinated and control groups in terms of proportion of C. burnetii shedders. These results suggest that optimal results of vaccination in heavily infected flocks may not be obtained in a short-term period.This work was supported by funding from INIA, FAU2006-00002-C04-01 and FEDER. Ianire Astobiza is the recipient of a predoctoral fellowship of the Department of Environment, Territorial Planning, Agriculture and Fisheries of the Basque Government. F. Ruiz-Fons is supported by a postdoctoral contract from the ‘Instituto de Salud Carlos III’ of the Spanish Government.Peer Reviewe

Measuring antibody levels in bulk-tank milk as an epidemiological tool to search for the status of Coxiella burnetii in dairy sheep

Pooling samples may provide a valuable alternative to individual testing for pathogen surveillance purposes. We studied the reliability of measuring the level of antibodies against Coxiella burnetii in bulk-tank milk (BTM) to estimate the seroprevalence of C. burnetii in dairy sheep in 34 flocks. We then estimated the seroprevalence of C. burnetii in 154 dairy sheep flocks according to the level of antibodies in BTM. We tested for the accuracy of our estimation at the population level by comparing predicted mean C. burnetii flock seroprevalence with that obtained in another survey performed on the same population. Our findings showed that testing BTM by ELISA is a cost-effective and relatively good index of the seroprevalence of C. burnetii in dairy sheep and may be a useful tool for epidemiological surveillance at the population level.This work was supported by funding from INIA, FAU2006-00002-C04-01 and FEDER. F. Ruiz-Fons is supported by the Instituto de Salud Carlos III of the Spanish Ministry of Science and Innovation. Ianire Astobiza is the recipient of a predoctoral fellowship of the Department of Agriculture, Fisheries and Food of the Basque Government.Peer Reviewe

Molecular investigation of the occurrence of Coxiella burnetii in wildlife and ticks in an endemic area

At present few studies have been carried out on the distribution and incidence of Coxiella burnetii infection in wildlife. Therefore, the aim of this study was to investigate the distribution of C. burnetii in the main wild species in the Basque Country (Northern Spain), such as carnivores, cervids, wild boar, lagomorphs and several species of birds. Tissues from a total of 601 animals and 340 adult ticks collected from them were analyzed by PCR. DNA of C. burnetii was detected in 5.1% of roe deer (Capreolus capreolus), 4.3% of wild boar (Sus scrofa), 9.1% of European hare (Lepus europaeus), and among wild birds, in 11% of vultures (Gyps fulvus) and 14% of black kites (Milvus migrans). These results showed that C. burnetii circulates in wildlife in Spain participating in the cycle of Q fever in nature. All of the adult ticks analyzed were negative for C. burnetii, suggesting that ticks do not play an important role in the transmission of C. burnetii in this area.This work was supported by funding from INIA, FAU2006-00002-C04-01, MECAG/2001-3947-C02-02, and FEDER. Ianire Astobiza is the recipient of a predoctoral fellowship of the Department of Agriculture, Fisheries and Food of the Basque Government. Francisco Ruiz-Fons is supported by the ‘Instituto de Salud Carlos III’ of the Spanish Ministry of Health.Peer Reviewe