HDAC class I inhibitor domatinostat sensitizes pancreatic cancer to chemotherapy by targeting cancer stem cell compartment via FOXM1 modulation

Pancreatic ductal adenocarcinoma (PDAC) represents an unmet clinical need due to the very poor prognosis and the lack of effective therapy. Here we investigated the potential of domatinostat (4SC-202), a new class I histone deacetylase (HDAC) inhibitor, currently in clinical development, to sensitize PDAC to first line standard gemcitabine (G)/taxol (T) doublet chemotherapy treatment. Methods: Synergistic anti-tumor effect of the combined treatment was assessed in PANC1, ASPC1 and PANC28 PDAC cell lines in vitro as well as on tumor spheroids and microtissues, by evaluating combination index (CI), apoptosis, clonogenic capability. The data were confirmed in vivo xenograft models of PANC28 and PANC1 cells in athymic mice. Cancer stem cells (CSC) targeting was studied by mRNA and protein expression of CSC markers, by limiting dilution assay, and by flow cytometric and immunofluorescent evaluation of CSC mitochondrial and cellular oxidative stress. Mechanistic role of forkhead box M1 (FOXM1) and downstream targets was evaluated in FOXM1-overexpressing PDAC cells. Results: We showed that domatinostat sensitized in vitro and in vivo models of PDAC to chemotherapeutics commonly used in PDAC patients management and particularly to GT doublet, by targeting CSC compartment through the induction of mitochondrial and cellular oxidative stress. Mechanistically, we showed that domatinostat hampers the expression and function of FOXM1, a transcription factor playing a crucial role in stemness, oxidative stress modulation and DNA repair. Domatinostat reduced FOXM1 protein levels by downregulating mRNA expression and inducing proteasome-mediated protein degradation thus preventing nuclear translocation correlated with a reduction of FOXM1 target genes. Furthermore, by overexpressing FOXM1 in PDAC cells we significantly reduced domatinostatinducing oxidative mitochondrial and cellular stress and abolished GT sensitization, both in adherent and spheroid cells, confirming FOXM1 crucial role in the mechanisms described. Finally, we found a correlation of FOXM1 expression with poor progression free survival in PDAC chemotherapy-treated patients

Phase 1/2 study of valproic acid and short-course radiotherapy plus capecitabine as preoperative treatment in low-moderate risk rectal cancer-V-shoRT-R3 (Valproic acid--short Radiotherapy--rectum 3rd trial).

BACKGROUND: Locally advanced rectal cancer (LARC) is a heterogeneous group of tumors where a risk-adapted therapeutic strategy is needed. Short-course radiotherapy (SCRT) is a more convenient option for LARC patients than preoperative long-course RT plus capecitabine. Histone-deacetylase inhibitors (HDACi) have shown activity in combination with RT and chemotherapy in the treatment of solid tumors. Valproic acid (VPA) is an anti-epileptic drug with HDACi and anticancer activity. In preclinical studies, our group showed that the addition of HDACi, including VPA, to capecitabine produces synergistic antitumour effects by up-regulating thymidine phosphorylase (TP), the key enzyme converting capecitabine to 5-FU, and by downregulating thymidylate synthase (TS), the 5-FU target. METHODS/DESIGN: Two parallel phase-1 studies will assess the safety of preoperative SCRT (5 fractions each of 5 Gy, on days 1 to 5) combined with (a) capecitabine alone (increasing dose levels: 500-825 mg/m2/bid), on days 1-21, or (b) capecitabine as above plus VPA (oral daily day -14 to 21, with an intra-patient titration for a target serum level of 50-100 microg/ml) followed by surgery 8 weeks after the end of SCRT, in low-moderate risk RC patients. Also, a randomized phase-2 study will be performed to explore whether the addition of VPA and/or capecitabine to preoperative SCRT might increase pathologic complete tumor regression (TRG1) rate. A sample size of 86 patients (21-22/arm) was calculated under the hypothesis that the addition of capecitabine or VPA to SCRT can improve the TRG1 rate from 5% to 20%, with one-sided alpha = 0.10 and 80% power.Several biomarkers will be evaluated comparing normal mucosa with tumor (TP, TS, VEGF, RAD51, XRCC1, Histones/proteins acetylation, HDAC isoforms) and on blood samples (polymorphisms of DPD, TS, XRCC1, GSTP1, RAD51 and XRCC3, circulating endothelial and progenitors cells; PBMCs-Histones/proteins acetylation). Tumor metabolism will be measured by 18FDG-PET at baseline and 15 days after the beginning of SCRT. DISCUSSION: This project aims to improve the efficacy of preoperative treatment of LARC and to decrease the inconvenience and the cost of standard long-course RT. Correlative studies could identify both prognostic and predictive biomarkers and could add new insight in the mechanism of interaction between VPA, capecitabine and RT.EudraCT Number: 2012-002831-28. TRIAL REGISTRATION: ClinicalTrials.gov number, NCT01898104

Analisi funzionale e contestualizzazione della ceramica in argilla grezza. Modalità di preparazione e di cottura degli alimenti nell’area flegrea tra l’età Orientalizzante e l’età Ellenistica.

Il lavoro della dott.ssa Serena Avallone affronta una problematica relativa allo studio della ceramica in argilla grezza attestata nei siti flegrei di Pithecusa e di Cuma in un arco cronologico compreso tra l’Orientalizzante Antico e l’Età Ellenistica. L’analisi di questa classe ceramica è stato rivolto al raggiungimento di una conoscenza esaustiva del repertorio vascolare partendo dalla ricostruzione delle caratteristiche tecniche e morfologiche, attraverso una classificazione delle argille e una scansione tipologica delle forme, avvalendosi dei nuovi approcci sperimentati dalla ricerca archeologica allo studio della classe quali quello dell’analisi funzionale e quello della contestualizzazione

Synergistic antitumor interaction between valproic acid, capecitabine and radiotherapy in colorectal cancer: critical role of p53

Abstract Background Recurrence with distant metastases has become the predominant pattern of failure in locally advanced rectal cancer (LARC), thus the integration of new antineoplastic agents into preoperative fluoropyrimidine-based chemo-radiotherapy represents a clinical challenge to implement an intensified therapeutic strategy. The present study examined the combination of the histone deacetylase inhibitor (HDACi) valproic acid (VPA) with fluoropyrimidine-based chemo-radiotherapy on colorectal cancer (CRC) cells. Methods HCT-116 (p53-wild type), HCT-116 p53−/− (p53-null), SW620 and HT29 (p53-mutant) CRC cell lines were used to assess the antitumor interaction between VPA and capecitabine metabolite 5′-deoxy-5-fluorouridine (5′-DFUR) in combination with radiotherapy and to evaluate the role of p53 in the combination treatment. Effects on proliferation, clonogenicity and apoptosis were evaluated, along with γH2AX foci formation as an indicator for DNA damage. Results Combined treatment with equipotent doses of VPA and 5′-DFUR resulted in synergistic effects in CRC lines expressing p53 (wild-type or mutant). In HCT-116 p53−/− cells we observed antagonist effects. Radiotherapy further potentiated the antiproliferative, pro-apoptotic and DNA damage effects induced by 5′-DFUR/VPA combination in p53 expressing cells. Conclusions These results highlighted the role of VPA as valuable candidate to be added to preoperative chemo-radiotherapy in LARC. On these bases we launched the ongoing phase I/II study of VPA and short-course radiotherapy plus capecitabine as preoperative treatment in low-moderate risk rectal cancer (V-shoRT-R3)

Gamma-Glutamyl Transferase Activity in Kids Born from Goats Fed Genetically Modified Soybean

The majority of animal feeding trials using GM feeds indicated no clinical effects while as concerns histopathological abnormalities in organs or tissues of exposed animals, the results are conflicting. Several data indicate liver and kidney problems as end points of GM diet effects. In rabbit fed GM soybean, it was hypothesised that cell metabolism of sev- eral enzymes was altered as well as elevated levels of LDH were revealed in tissues and organs of kids when mothers are fed GM soybean. The objective of this study was to investigate the fate of transgenic DNA and the activity of gamma-glutamyl transferase (GGT) in blood, liver and kidney from kids fed only milk of their mother fed conventional (control) or genetically modified soybean meal solvent extract (treated). PCR analysis revealed that fragments of mul- ticopy chloroplast (trnL) and single soybean-specific (lectin) gene were found in samples of both groups. Fragments of transgenic gene were found only in treated kids: detection of 35S promoter was significant in liver, kidney and blood, and detection of CP4 epsps gene fragment was significant in liver and kidney. Concerning GGT, no differences were found in serum, while its activity was significantly (P < 0.01) higher in kidney (63.4 ± 7.8 vs 81.2 ± 11.3 u/g of tissue) and liver (40.1 ± 5.2 vs 62.6 ± 9.8 u/g of tissue) of kids from treated goats. The increase in GGT activity was confirmed by histochemistry

Diet Aloe supplementation in pregnant buffalo cows improves colostrum immunoglobulin content.

Colostrum ingestion in ruminants is the one way to get maternal antibodies. Calves start suckling soon after birth, and the absorption of immunoglobulin (Ig) lasts up to 24 hours. Many diseases of neonatal calves are related to a poor colostrum quality, which is determined by the content of IgG. The polysaccharide fractions of Aloe has been reported as potent B cell stimulators either in vitro or in vivo studies. Aim of the present research was to evaluate the influence of Aloe arborescence fed to pregnant dry buffalo cows on IgG concentration of colostrum. Twenty-four pluriparae buffalo cows were divided into two homogeneous groups during the last two months of pregnancy. Group A received a supplementation of 50g/day/head of a commercial product containing Aloe arborescence, while group B was the control. Within one hour from calving, samples of colostrum were collected from each subject and underwent IgG assay. A significant (P<0.05) increase of colostrum IgG concentration was detected in colostrum from dams supplemented with Aloe. Results showed that Aloe supplementation can increase the immunological properties of colostrum thus resulting in improving passive transfer in newborn calves

In vitro toxicity of fibrous glaucophane

The health hazard represented by the exposure to asbestos may also concern other minerals with asbestos-like crystal habit. One of these potentially hazardous minerals is fibrous glaucophane. Fibrous glaucophane is a major component of blueschist rocks of California (USA) currently mined for construction purposes. Dust generated by the excavation activities might potentially expose workers and the general public. The aim of this study was to determine whether fibrous glaucophane induces in vitro toxicity effects on lung cells by assessing the biological responses of cultured human pleural mesothelial cells (Met-5A) and THP-1 derived macrophages exposed for 24h and 48h to glaucophane fibres. Crocidolite asbestos was tested for comparison. The experimental configuration of the in vitro tests included a cell culture without fibres (i.e., control), cell cultures treated with 50g/mL (i.e., 15.6g/cm2) of crocidolite fibres and 25-50-100g/mL (i.e., 7.8-15.6-31.2g/cm2) of glaucophane fibres. Results showed that fibrous glaucophane may induce a decrease in cell viability and an increase in extra-cellular lactate dehydrogenase release in the tested cell cultures in a concentration dependent mode. Moreover, it was found that fibrous glaucophane has a potency to cause oxidative stress. The biological reactivity of fibrous glaucophane confirms that this mineral fibre is a toxic agent and, although it apparently induces lower toxic effects compared to crocidolite, exposure to its fibre may be responsible for the development of lung diseases in exposed unprotected workers and population