The role of the WISE Consortium in the European DEDIPAC-KH project

WISE (Wellness, nutrItion, Sport and Exercise prevention) is a research consortium including five Italian research teams (University of Turin, University of Milan, University of Trieste, Universiy of Rome “Foro Italico”, University of Bari), operating within the broader framework of the DEDIPAC-KH joint action (Determinants of Diet and Physical Activity Knowledge Hub). Research actions within the WISE consortium, funded by the Italian Ministry of Higher Education & Research, are in line with the main objective of the DEDIPAC-KH of developing an international and interdisciplinary network of researchers on dietary, physical activity and sedentary behaviours, related determinant research and policy interventions. More specifically, the WISE consortium research aimed to contribute to the following task (1.2.4 - Task Leader: Prof. Alan Donnelly): perform SLRs to identify state-of the art methods for physical activity and sedentary behaviour measurements. The focus of task 1.2.4 was to examine the methodological effectiveness (validity, reliability and sensitivity/responsiveness) of measures of physical activity and sedentary behaviours. The approach taken with this task was to examine the methodological effectiveness of measures of physical activity and sedentary behaviours in two populations; i) child/adolescence and ii) adults. Findings on methodological effectiveness of measures of physical activity and sedentary behaviours constitute the basis for a variety of publication and reports, and conference communications. The DEDIPAC-KH project created an unique opportunity for developing a comprehensive analysis on the determinants of diet and physical activity in Italy, and fostered successful collaboration with leading international groups. The findings of the WISE project created valuable information for the implementation of successful policies in Italy.This work was supported by MIUR

Synthesis and bioactivity of novel histone acetylation inhibitors : potential new drugs for treatment of cancer and inflammation

There is an increasing interest in histone acetyltransferases (HATs) as new therapeutic targets for treatment of diseases like, for example, inflammation and cancer. However, only few small and cell-permeable molecule inhibitors of HATs are currently available. The work described in this thesis focuses on the synthesis and biological evaluation of novel HAT inhibitors. The major challenge is to find potent and selective HAT inhibitors that show biological activity in cells. In the first part of this thesis we report the design and synthesis of potent isothiazolone-based inhibitors of the recombinant HAT PCAF, which are also cytotoxic for different cancer cell lines. Moreover, we elucidate the chemical reactivity of isothiazolones derivatives showing that isothiazolones might find applications in activity-based protein profiling studies. The second part of this thesis describes the development of novel HAT inhibitors using the natural product anacardic acid (AA) as starting point. Using a combination of molecular design, organic synthesis and enzyme inhibition studies, we develop several novel compounds that are selective inhibitors of the recombinant HAT Tip60 in comparison with the HATs PCAF and p300. Furthermore, we identify a new AA derivative that shows a twofold improved potency for inhibition of histone H4 acetylation in cell-based studies. In conclusion, the work described in this thesis provides new chemical tools for pharmacological studies to validate HATs inhibitors as potential new drugs for treatment of cancer and inflammation

Reactivity of isothiazolones and isothiazolone-1-oxides in the inhibition of the PCAF histone acetyltransferase

Development of small molecule inhibitors of the histone acetyltransferase p300/CBP associated factor (PCAF) is relevant for oncology. The inhibition of the enzyme PCAF and proliferation of the cancer cell line HEP G2 by a series of 5-chloroisothiazolones was compared to a series of 5-chloroisothiazolone-1-oxides. The PCAF inhibitory potency of 5-chloroisothiazolones and 5-chloroisothiazolone-1-oxides is influenced by substitution in the 4-position. A study on the reactivity of the HAT inhibitors towards thiols and thiolates indicates that 5-chloroisothiazolones reacted quickly with propane-1-thiolate to provide many products, whereas 5-chloroisothiazolone-1-oxides provide only one defined product. Growth inhibition studies indicate that 5-chloroisothiazolones inhibit proliferation of HEP G2 cells at concentrations between 8.6 and 24 mu M, whereas 5-chloroisothiazolone-1-oxides required higher concentrations or showed no inhibition. (C) 2009 Elsevier Masson SAS. All rights reserved

Histone acetyltransferases are crucial regulators in NF-kappa B mediated inflammation

Post-translational modifications of proteins, such as acetylation, are important regulatory events in eukaryotic cells. Reversible acetylations of histones and non-histone proteins regulate gene expression and protein activity. Acetylation levels of proteins are regulated by a dynamic equilibrium between acetylation by (histone) acetyltransferases and deacetylation by (histone) deacetylases. Alterations in this equilibrium can result in pathological states. Inflammation is a physiological response that, under certain conditions, turns into a disease. This review focuses on the crucial regulatory roles of protein acetylation in NF-kappa B-mediated inflammation and the potential applications of small-molecule inhibitors of acetylation for the treatment of inflammatory diseases

Anacardic acid derived salicylates are inhibitors or activators of lipoxygenases

Lipoxygenases catalyze the oxidation of unsaturated fatty acids, such as linoleic acid, which play a crucial role in inflammatory responses. Selective inhibitors may provide a new therapeutic approach for inflammatory diseases. In this study, we describe the identification of a novel soybean lipoxygenase-1 (SLO-1) inhibitor and a potato 5-lipoxygenase (5-LOX) activator from a screening of a focused compound collection around the natural product anacardic acid. The natural product anacardic acid inhibits SLO-1 with an IC50 of 52 mu M, whereas the inhibitory potency of the novel mixed type inhibitor 23 is fivefold enhanced. In addition, another derivative (21) caused non-essential activation of potato 5-LOX. This suggests the presence of an allosteric binding site that regulates the lipoxygenase activity. (C) 2012 Elsevier Ltd. All rights reserved

Inhibition of PCAF by Anacardic Acid Derivative Leads to Apoptosis and Breaks Resistance to DNA Damage in BCR-ABL-expressing Cells

Acetylation of histones and nonhistone proteins is a posttranslational modification which plays a major role in the regulation of intracellular processes involved in tumorigenesis. It was shown that different acetylation of proteins correlates with development of leukemia. It is proposed that histone acetyltransferases (HATs) are important novel drug targets for leukemia treatment, however data are still not consistent. Our previous data showed that a derivative of anacardic acid - small molecule MG153, which has been designed and synthesized to optimize the HAT inhibitory potency of anacardic acid, is a potent inhibitor of p300/CBP associated factor (PCAF) acetyltransferase. Here we ask whether inhibition of PCAF acetyltransferase with MG153 will show proapoptotic effects in cells expressing BCR-ABL, which show increased PCAF expression and are resistant to apoptosis. We found that inhibition of PCAF decreases proliferation and induces apoptosis, which correlates with loss of the mitochondrial membrane potential and DNA fragmentation. Importantly, cells expressing BCR-ABL are more sensitive to PCAF inhibition compared to parental cells without BCR-ABL. Moreover, inhibition of PCAF in BCR-ABL-expressing cells breaks their resistance to DNA damage-induced cell death. These findings provide direct evidence that targeting the PCAF alone or in combination with DNA-damaging drugs shows cytotoxic effects and should be considered as a prospective therapeutic strategy in chronic myeloid leukemia cells. Moreover, we propose that anacardic acid derivative MG153 is a valuable agent and further studies validating its therapeutic relevance should be performed