C-Met/miR-130b axis as novel mechanism and biomarker for castration resistance state acquisition.

Although a significant subset of prostate tumors remain indolent during the entire life, the advanced forms are still one of the leading cause of cancer-related death. There are not reliable markers distinguishing indolent from aggressive forms. Here we highlighted a new molecular circuitry involving microRNA and coding genes promoting cancer progression and castration resistance. Our preclinical and clinical data demonstrated that c-Met activation increases miR-130b levels, inhibits androgen receptor expression, promotes cancer spreading and resistance to hormone ablation therapy. The relevance of these findings was confirmed on patients' samples and by in silico analysis on an independent patient cohort from Taylor's platform. Data suggest c-Met/miR-130b axis as a new prognostic marker for patients' risk assessment and as an indicator of therapy resistance. Our results propose new biomarkers for therapy decision-making in all phases of the pathology. Data may help identify high-risk patients to be treated with adjuvant therapy together with alternative cure for castration-resistant forms while facilitating the identification of possible patients candidates for anti-Met therapy. In addition, we demonstrated that it is possible to evaluate Met/miR-130b axis expression in exosomes isolated from peripheral blood of surgery candidates and advanced patients offering a new non-invasive tool for active surveillance and therapy monitoring.

A microRNA code for prostate cancer metastasis.

Although the development of bone metastasis is a major detrimental event in prostate cancer, the molecular mechanisms responsible for bone homing and destruction remain largely unknown. Here we show that loss of miR-15 and miR-16 in cooperation with increased miR-21 expression promote prostate cancer spreading and bone lesions. This combination of microRNA endows bone-metastatic potential to prostate cancer cells. Concomitant loss of miR-15/miR-16 and gain of miR-21 aberrantly activate TGF-ß and Hedgehog signaling, that mediate local invasion, distant bone marrow colonization and osteolysis by prostate cancer cells. These findings establish a new molecular circuitry for prostate cancer metastasis that was validated in patients' cohorts. Our data indicate a network of biomarkers and druggable pathways to improve patient treatment.

BTG2 loss and miR-21 upregulation contribute to prostate cell transformation by inducing luminal markers expression and epithelial-mesenchymal transition.

Prostate cancer is one of the leading causes of cancer-related death in men. Despite significant advances in prostate cancer diagnosis and management, the molecular events involved in the transformation of normal prostate cells into cancer cells have not been fully understood. It is generally accepted that prostate cancer derives from the basal compartment while expressing luminal markers. We investigated whether downregulation of the basal protein B-cell translocation gene 2 (BTG2) is implicated in prostate cancer transformation and progression. Here we show that BTG2 loss can shift normal prostate basal cells towards luminal markers expression, a phenotype also accompanied by the appearance of epithelial-mesenchymal transition (EMT) traits. We also show that the overexpression of microRNA (miR)-21 suppresses BTG2 levels and promotes the acquisition of luminal markers and EMT in prostate cells. Furthermore, by using an innovative lentiviral vector able to compete with endogenous mRNA through the overexpression of the 3'-untranslated region of BTG2, we demonstrate that in prostate tumor cells, the levels of luminal and EMT markers can be reduced by derepression of BTG2 from microRNA-mediated control. Finally, we show that the loss of BTG2 expression confers to non-tumorigenic prostate cells ability to grow in an orthotopic murine model, thus demonstrating the central role of BTG2 downregulaton in prostate cancer biology.

Systemic in vivo lentiviral delivery of miR-15a/16 reduces malignancy in the NZB de novo mouse model of chronic lymphocytic leukemia.

Similar to human chronic lymphocytic leukemia (CLL), the de novo New Zealand Black (NZB) mouse model has a genetically determined age-associated increase in malignant B-1 clones and decreased expression of microRNAs miR-15a and miR-16 in B-1 cells. In the present study, lentiviral vectors were employed in vivo to restore miR-15a/16, and both the short-term single injection and long-term multiple injection effects of this delivery were observed in NZB. Control lentivirus without the mir-15a/16 sequence was used for comparison. We found that in vivo lentiviral delivery of mir-15a/16 increased miR-15a/16 expression in cells that were transduced (detected by GFP expression) and in sera when compared with control lentivirus treatment. More importantly, mice treated with the miR-expressing lentivirus had decreased disease. The lentivirus had little systemic toxicity while preferentially targeting B-1 cells. Short-term effects on B-1 cells were direct effects, and only malignant B-1 cells transduced with miR-15a/16 lentivirus had decreased viability. In contrast, long-term studies suggested both direct and indirect effects resulting from miR-15a/16 lentivirus treatment. A decrease in B-1 cells was found in both the transduced and non-transduced populations. Our data support the potential use of systemic lentiviral delivery of miR-15a/16 to ameliorate disease manifestations of CLL.

Control of tumor and microenvironment cross-talk by miR-15a and miR-16 in prostate cancer.

The interaction between cancer cells and microenvironment has a critical role in tumor development and progression. Although microRNAs regulate all the major biological mechanisms, their influence on tumor microenvironment is largely unexplored. Here, we investigate the role of microRNAs in the tumor-supportive capacity of stromal cells. We demonstrated that miR-15 and miR-16 are downregulated in fibroblasts surrounding the prostate tumors of the majority of 23 patients analyzed. Such downregulation of miR-15 and miR-16 in cancer-associated fibroblasts (CAFs) promoted tumor growth and progression through the reduced post-transcriptional repression of Fgf-2 and its receptor Fgfr1, which act on both stromal and tumor cells to enhance cancer cell survival, proliferation and migration. Moreover, reconstitution of miR-15 and miR-16 impaired considerably the tumor-supportive capability of stromal cells in vitro and in vivo. Our data suggest a molecular circuitry in which miR-15 and miR-16 and their correlated targets cooperate to promote tumor expansion and invasiveness through the concurrent activity on stromal and cancer cells, thus providing further support to the development of therapies aimed at reconstituting miR-15 and miR-16 in advanced prostate cancer.

[Quality of life assessment: validation of the Italian version of the WHOQOL-Brief]. / La valutazione della qualità della vita: validazione del WHOQOL-Breve.

OBJECTIVE: To test the psychometric properties of the Italian version of the WHOQOL-BRIEF (e.g., construct and internal validity, concurrent validity with the MOS SF-36 and test-retest reliability). The WHOQOL-BRIEF is a 26-items self-report instrument which assesses four domains assumed to represent the Quality Of Life (QOL) construct: physical domain, psychological domain, social relationships domain and environment domain, plus two facets for assessing overall QOL and general health. METHODS: Data have been collected in three sites (Bologna, Modena and Padua), located in the North of Italy, in the framework of the international WHOQOL project. According to the study design, the sample had to include about 50% males and 50% females, 50% of subjects below and 50% above the age of 45, all in contact with various health services. A subsample has been re-interviewed after 2-3 weeks in order to study test-retest reliability. After the WHOQOL-BRIEF, most subjects have also been administered the MOS-SF36 in order to test the concurrent validity between these two instruments. RESULTS: The instrument was administered to 379 subjects (1/6 healthy and 1/6 sick), chosen to be representative of a variety of different medical conditions. Seventy patients, who displayed stable health conditions, have been reassessed after 2-3 weeks to study test-retest reliability. The WHOQOL-BRIEF domains has shown good internal consistency, ranging from 0.65 for the social relationships domain to 0.80 for the physical domain; it has been able to discriminate between in- and out-patients and between the two age groups considered in the present study (< 45, > or = 45 years). Only physical and psychological domains were found to discriminate between healthy and ill subjects. No gender differences in the mean scores for the four domains were found. Concurrent validity between the WHOQOL-Brief and the MOS-SF-36 was satisfactory, and specific for the physical and psychological health domains. Test-retest reliability values were also good, ranging from 0.76 for the environment domain to 0.93 for the psychological domain. CONCLUSIONS: This study shows that the WHOQOL-BRIEF is psychometrically valid and reliable, and that it is also potentially useful in discriminating between subjects with different health conditions in clinical settings.

Prevalence of HBV markers in the Italian Armed Forces and HBV vaccination.

Hepatitis B (HBV) markers were studied in 710 subjects who had been on active duty for over 6 months in the Italian Armed Forces. The prevalence of HBsAg carriers was found to be 4.4%, while 31.6% of the subjects were positive for various HBV antibodies. A total of 137 subjects were vaccinated with an anti-HBV vaccine (HB-VAX, MSD). The percentages of non-responders and low responders were 13.86% and 13.14%, respectively. Boosters administered 3 months post-vaccination schedule, with or without immunostimulatory treatment, resulted in seroconversions and/or substantial increases in HBsAb levels in 50% of these subjects.

Cellular and humoral modifications during response to HBsAg vaccine in healthy subjects: comparison between French and American vaccines.

Two groups of 18 healthy adults, chosen from a risk population, were vaccinated against hepatitis B virus with French (Hevac B, Pasteur) or American (HB-VAX, MSD) vaccine; immunological and biochemical parameters (HBsAb titers, Ig, IgM-rheumatoid factor (IgM-RF) levels, autologous mixed lymphocyte reaction (AMLR), T4/T8 ratio, lymphocyte migration inhibition test (LMIT) with specific antigen and transaminase levels) were compared at preselected time intervals. The vaccines were found to be clinically safe, as documented by the absence of demonstrable side effects and changes in serum transaminase levels. The positivity of the LMIT test demonstrated an early immunization on day 45 in all subjects tested, along with a marked reduction in AMLR, independent of the vaccine used. A slightly higher, though not statistically significant, percentage of seroconversion was observed for the Pasteur vaccine. This same preparation induced a significant increase in the main Ig classes and in Ig;-RF levels on day 45. This data indicates that aspecific modification of the immune system occur to a greater extent with Hevac B, presumably for its lower degree of purification. Thus, despite the slightly lower efficacy, the HB-VAX might be the preparation of choice in subjects with immunological disturbances.

Comparative analysis of immunological responses to oral (Ty21a) and parenteral (TAB) typhoid vaccines.

The clinical and immunological responses to typhoid vaccination with parenteral (TAB) and oral (Ty21a) vaccines in two groups of 30 adult male subjects were studied. Parameters monitored included specific anti-Salmonella typhi cell-mediated immunity and total and specific antilipopolysaccharide fecal immunoglobulin A (IgA) titers in Ty21a-vaccinated subjects. Peripheral blood lymphocytes antibacterial activity was significantly increased only in Ty21a-vaccinated subjects. Serum arming activity and results of human F(ab')2 anti-IgG and -IgA inhibition tests suggest antibody-dependent cellular cytotoxicity mediated by IgA in those vaccinated with Ty21a. Interestingly enough, the cells of TAB-vaccinated subjects were able to mediate IgG-dependent cellular cytotoxicity, as was observable from the results of blocking experiments. Moreover, total and specific antilipopolysaccharide fecal IgA levels were observed to be significantly increased with Ty21a, up to 8 months post-vaccination schedule. An early-onset, transitory increase in serum IgM rheumatoid factor was also found, exclusively in subjects treated with TAB, and was no longer detectable on day 240. Ty21a was well tolerated and free of side effects, whereas 65% of subjects administered TAB reported fever, headache, malaise, and local tenderness at the injection site. Our data show that the two typhoid vaccines induce different cell-mediated specific immune responses. The role of these responses in protection against Salmonella infection, however, requires further investigation.

A subchronic dermal exposure study of diethylene glycol monomethyl ether and ethylene glycol monomethyl ether in the male guinea pig.

Diethylene glycol monomethyl ether (DEGME) has been selected as a replacement anti-icing additive for ethylene glycol monomethyl ether (EGME) in Navy jet aircraft fuel. This experiment was performed to determine whether DEGME produced similar toxicity to EGME following dermal exposure. Male guinea pigs were dermally exposed to 1.00, 0.20, 0.04, or 0 (control) g/kg/day DEGME for 13 weeks, 5 days/week, 6 hr/day. Another group of animals was similarly exposed to 1.00 g/kg/day EGME. Body weights as well as testicular and splenic weights were reduced as a result of exposure to EGME, DEGME-exposed animals exhibited decreased splenic weight in the high- and medium-dose (1.00 and 0.20 g/kg/day) exposure groups only. Hematologic changes in EGME-exposed animals included mild anemia with increased erythrocytic mean corpuscular volumes and a lymphopenia with increased neutrophils. Similar hematological changes were not observed in any animals exposed to DEGME. Serum creatine kinase activity was increased in animals exposed to EGME, and serum lactate dehydrogenase activity was increased in EGME and 1.00 g/kg/day DEGME-exposed animals. In general, DEGME produced minimal toxicological changes following dermal exposure, whereas the toxicological changes observed following similar exposure to EGME were much more profound.

Pulmonary alveolar macrophage response to hyperbaric carbon monoxide exposure.

The sublethal effects of carbon monoxide (CO) in hyperbaric environments have not been adequately characterized. A physiologically sensitive indicator of sublethal effects of a contaminant is the pulmonary alveolar macrophage (PAM). The guinea pig PAM and its reaction to various CO concentrations at 8 atmospheres absolute (ATA) and the surface-equivalent concentrations at 1 ATA were studied in this experiment. In the absence of CO, PAM counts and viabilites from guinea pigs exposed to a helium-oxygen environment at either 1 or 8 ATA were not significantly different from one another. Mean PAM viability for guinea pigs exposed to CO concentrations at 2250--4200 mg/m3 at 8 ATA was 68.4 +/- 7.3% and was not significantly different from the 1-ATA viability value of 72.0 +/- 4.5% for the 1600--4200 mg/m3 CO range. The data show that at 1 to 8 ATA and CO concentrations of 1600--4200 mg/m3, the decrease in PAM viability was accompanied by dramatic five- to sixfold increases in PAM counts.