Defective mitosis with chromosome missegregation can have a dramatic effect on genome integrity by causing DNA damage, activation of the DNA damage response (DDR), and chromosomal instability. ...Although this is an energy-dependent process, mechanisms linking DDR to cellular metabolism are unknown. Here we show that checkpoint kinase 2 (CHK2), a central effector of DDR, regulates cellular energy production by affecting glycolysis and mitochondrial functions. Patients with hepatocellular carcinoma (HCC) had increased
mRNA in blood, which was associated with elevated tricarboxylic acid cycle (TCA) metabolites. CHK2 controlled expression of succinate dehydrogenase (SDH) and intervened with mitochondrial functions. DNA damage and CHK2 promoted SDH activity marked by increased succinate oxidation through the TCA cycle; this was confirmed in a transgenic model of HCC with elevated DNA damage. Mitochondrial analysis identified CHK2-controlled expression of SDH as key in sustaining reactive oxygen species production. Cells with DNA damage and elevated CHK2 relied significantly on glycolysis for ATP production due to dysfunctional mitochondria, which was abolished by
knockdown. This represents a vulnerability created by the DNA damage response that could be exploited for development of new therapies. SIGNIFICANCE: This study uncovers a link between a central effector of DNA damage response, CHK2, and cellular metabolism, revealing potential therapeutic strategies for targeting hepatocellular carcinoma.
Abstract
Hepatocellular carcinoma (HCC) is nowadays the sixth cause of tumour-related deceases worldwide, estimated to become the third in Western countries by 2030. New drugs for HCC treatment still ...have many adverse effects. Several lines of evidence indicate that plant metabolites offer concrete opportunities for developing new therapeutic strategies for many diseases, including cancer. We previously reported that ethyl acetate extract of a spontaneous edible plant harvested in Apulia,
Crithmum maritimum
, significantly inhibited cell growth in HCC cells. By
1
H-NMR spectroscopy, here we show that
Crithmum maritimum
ethyl acetate extract counteracts the Warburg effect, by reducing intracellular lactate, inhibits protein anabolism, by decreasing amino acid level, and affects membrane biosynthesis by lowering choline and phosphocholine. Also, we observed an effect on lipid homeostasis, with a reduction in triglycerides, cholesterol, monounsaturated fatty acids (MUFA), and diunsaturated fatty acids (DUFA), and an increase in polyunsaturated fatty acids (PUFA). Taken together, these data demonstrate that
Crithmum maritimum
-induced cytostasis is exerted through a multi-effect action, targeting key metabolic processes in HCC cells. Overall, our findings highlight the role of
Crithmum maritimum
as a promising tool for the prevention and the improvement of the therapeutic options for HCC and other types of tumours.
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A
H-NMR-based metabolomic study was performed on MCF-7 cell lines treated with a novel nicotinamide derivative (DT-8) in comparison with two drugs characterized by a well-established mechanism of ...action, namely the DNA-metalating drug cisplatin (cis-diamminedichloridoplatinum(II), CDDP) and the antimitotic drug vinblastine (vinblastine, VIN). The effects of the three compounds, each one at the concentration corresponding to the IC
value, were investigated, with respect to the controls (K), by the
H-NMR of cells lysates and multivariate analysis (MVA) of the spectroscopic data. Relevant differences were found in the metabolic profiles of the different treatments with respect to the controls. A large overlap of the metabolic profiles in DT-8 vs. K and VIN vs. K suggests a similar biological response and mechanism of action, significantly diverse with respect to CDDP. On the other hand, DT8 seems to act by disorganizing the mitotic spindle and ultimately blocking the cell division, through a mechanism implying methionine depletion and/or
adenosylmethionine (SAM) limitation.
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Crop production and natural resource use, especially in developing countries, represents one of the most important food sources for humans. In particular, two wheat species (tetraploid, which is ...mostly used for pasta and hexaploid, which is primarily used for bread) account for about 20% of the whole calories consumed worldwide. In order to assess the mineral accumulation capability of some popular tetraploid wheat genotypes, a metabolomic (metallomic) approach was used in this study. The metallomic profile related to micro- (Zn, Fe, Cu, Mn, Ni and Cr), macro- (Ca, Mg and K) and toxic trace elements (Cd and Pb) was obtained by ICP-AES analysis in a large set of tetraploid wheat genotypes (Triticum turgidum L.) that were grown in two different experimental fields. Correlations and multivariate statistical analyses were performed, grouping the samples under two wheat sets, comprising cultivated durum cultivars (T. turgidum subsp. durum) and wild accessions (T. turgidum subsp. dicoccum and subsp. dicoccoides). The site dependence ranking for the selected genotypes with the highest nutrient accumulation was obtained. The significantly higher content of Mg (among the macronutrients) and the highest levels of Mn, Fe and Zn (among the micronutrients) were found for wild accessions with respect to durum cultivars. Moreover, the former genotypes were also the ones with the lowest level of accumulation of the trace toxic elements, in particular Cd. According to the performed statistical analyses, the wild accessions appeared also to be less influenced by the different environmental conditions. This is in accord with literature data, indicating the superiority of “old” with respect to modern wheat cultivars for mineral content. Although further studies are required on a wider range of genotypes to confirm these findings, the obtained results could be used to better select the less demanding and better performing cultivars in specific target wheat growing environments.
•Moderate discrimination among samples by different vinifications was observed.•Macroparameters were not conditioned by soil management and training system.•PCA and OPLS-DA were able to discriminate ...according to different soil management.•Higher content of glycerol and aromatic compounds was reported for cover crop soil.
Metabolic profiles of 32 Negroamaro red wines were analysed using 1H NMR spectroscopy and multivariate statistical analyses (Principal Component Analysis, PCA, and Orthogonal Partial Least Square Discriminant Analysis, OPLS-DA). Among winemaking technologies three were compared: ultrasounds (U; 12 samples), cryomaceration using dry ice (C; 12 samples) and traditional (T; 8 samples). Moreover, each vinification technology was used for grapes grown by two different soil management practices, soil tillage (ST; 16 samples) and cover crop (CC; 16 samples), and by two different training systems, monolateral (M; 16 samples) and bilateral Guyot (B; 16 samples). All statistical models applied on NMR data revealed a good separation between ST (soil tillage) and CC (cover crop), showing a higher influence of the soil management practices compared to the winemaking technologies (ultrasound, cryomaceration and traditional). The differentiation among samples, due to soil management practices, was mainly caused by metabolites such as glycerol, 2,3-butanediol, malic acid, α/β-glucose and phenolic compounds, such as tyrosine and caffeic acid.
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•Comparison highlighted differences in raw NMR data sets from the two laboratories.•The amount of shared information was significantly high (almost 70%).•Classification performance was independent ...from operator and instrumentation.•The two data sets shared the same metabolites discerning between olive oil cultivars.•The two data sets were fused in a unique model with high classification ability.
Nuclear Magnetic Resonance (NMR) spectroscopy is emerging as a powerful technique in olive oil fingerprinting, but its analytical robustness has to be proved. Here, we report a comparative study between two laboratories on olive oil 1H NMR fingerprinting, aiming to demonstrate the robustness of NMR-based metabolomics in generating comparable data sets for cultivar classification. Sample preparation and data acquisition were performed independently in two laboratories, equipped with different resolution spectrometers (400 and 500MHz), using two identical sets of mono-varietal olive oils. Partial Least Squares (PLS)-based techniques were applied to compare the data sets produced by the two laboratories. Despite differences in spectrum baseline, and in intensity and shape of peaks, the amount of shared information was significant (almost 70%) and related to cultivar (same metabolites discriminated between cultivars). In conclusion, regardless of the variability due to operator and machine, the data sets from the two participating units were comparable for the purpose of classification.
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Jellyfish outbreaks in marine coastal areas represent an emergent problem worldwide, with negative consequences on human activities and ecosystem functioning. However, potential positive effects of ...jellyfish biomass proliferation may be envisaged as a natural source of bioactive compounds of pharmaceutical interest. We investigated the biochemical composition of mature female gonads and lysozyme antibacterial activity of oocytes in the Mediterranean barrel jellyfish
. Chemical characterization was performed by means of multinuclear and multidimensional NMR spectroscopy. The ovaries of
were mainly composed of water (93.7 ± 1.9% of wet weight), with organic matter (OM) and dry weight made respectively of proteins (761.76 ± 25.11 µg mg
and 45.7 ± 1.5%), lipids (192.17 ± 10.56 µg mg
and 9.6 ± 0.6%), and carbohydrates (59.66 ± 2.72 µg mg
and 3.7 ± 0.3%). The aqueous extract of
gonads contained free amino acids, organic acids, and derivatives; the lipid extract was composed of triglycerides (TG), polyunsaturated fatty acids (PUFAs), diunsaturated fatty acids (DUFAs), monounsaturated fatty acids (MUFAs), saturated fatty acids (SFAs), and minor components such as sterols and phospholipids. The
oocyte lysate exhibited an antibacterial lysozyme-like activity (mean diameter of lysis of 9.33 ± 0.32 mm corresponding to 1.21 mg/mL of hen egg-white lysozyme). The occurrence of defense molecules is a crucial mechanism to grant healthy development of mature eggs and fertilized embryos (and the reproductive success of the species) by preventing marine bacterial overgrowth. As a corollary, these results call for future investigations for an exploitation of
biomasses as a resource of bioactive metabolites of biotechnological importance including pharmaceuticals and nutrition.
The biological invasion of the green algae
represents a serious scientific and public issue in the Mediterranean Sea, essentially due to strong modifications both to habitat structure and native ...benthic communities. Although alterations in health status and changes in flesh quality of some marine species (dietary exposed to
) have been observed, no studies on cause-effect relationships have been carried out. Here, for the first time, through a controlled feeding experiment followed by ¹H NMR Spectroscopy and multivariate analysis (PCA, OPLS-DA), we showed that caulerpin taken with diet is directly responsible of changes observed in metabolic profile of fish flesh, including alteration of lipid metabolism, in particular with a reduction of ω3 PUFA content. The potential of caulerpin to directly modulate lipid metabolism opens up new questions about causal mechanism triggered by algal metabolite also in view of a possible exploitation in the nutraceutical/medical field.
Thanks to recent advances in analytical technologies and statistical capabilities, the application field of metabolomics has increased significantly. Currently, this approach is used to investigate ...biological substrates looking for metabolic profile alterations, diseases markers, and drug effects. In particular, NMR spectroscopy has shown great potential as a detection technique, mainly for the ability to detect multiple (10s to 100s) metabolites at once without separation. Only in recent years has the NMR-based metabolomic approach been extended to investigate the cell metabolic alterations induced by metal-based antitumor drug administration. As expected, these studies are mainly focused on platinum complexes, but some palladium and ruthenium compounds are also under investigation. The use of a metabolomics approach was very effective in assessing tumor response to drugs and providing insights into the mechanism of action and resistance. Therefore, metabolomics may open new perspectives into the development of metal-based drugs. In particular, it has been shown that NMR-based, in vitro metabolomics is a powerful tool for detecting variations of the cell metabolites induced by the metal drug exposure, thus offering also the possibility of identifying specific markers for in vivo monitoring of tumor responsiveness to anticancer treatments.
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Xylella fastidiosa subsp. pauca is associated with the olive quick decline syndrome in Salento (Apulia region, Southern Italy). The first outbreak of the disease was noticed in the Gallipoli ...district, from where it subsequently reached nearby areas. To date, no specific study has verified if abnormal soil or leaf micronutrients is associated with the disease. Soil and leaf samples were taken from 23 olive farms showing symptoms of the syndrome located in Gallipoli and nearby areas. Each sample was analyzed for magnesium and micronutrients content using inductively coupled plasma atomic emission spectroscopy. Real-time PCR indicated that X. f. subsp. pauca was present in each sampled tree. There is a general lack of molybdenum in soil and low bioavailability of copper and molybdenum in tree leaves. Low content of manganese in soil was also found in some farms located in Gallipoli, Galatone and Trepuzzi. Olive trees grown in Gallipoli area also had low content of boron. Principal component analysis showed that soil and leaf samples from this area had lower micronutrient contents compared with other areas. General copper depletion in leaves was uncommon and has not been previously recorded in Italy in a large area. This could indicate that X. f. subsp. pauca infection causes a depletion of copper within olive leaves. Reduced copper content has been previously recorded in leaves infected with X. f. subsp. multiplex and for other bacterial pathogens. The role of copper in relation to X. fastidiosa infection is discussed. Keywords. Olive Quick Decline Syndrome, quarantine bacteria, micronutrients, Inductively Coupled Plasma Atomic Emission Spectroscopy Analysis.
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