Organelle adaptations in response to mechanical forces during tumour dissemination.

Cell migration plays a pivotal role in various biological processes including cancer dissemination and successful metastasis, where the role of mechanical signals is increasingly acknowledged. This review focuses on the intricate mechanisms through which cancer cells modulate their migratory strategies via organelle adaptations in response to the extracellular matrix (ECM). Specifically, the nucleus and mitochondria emerge as pivotal mediators in this process. These organelles serve as sensors, translating mechanical stimuli into rapid metabolic alterations that sustain cell migration. Importantly, prolonged exposure to such stimuli can induce transcriptional or epigenetic changes, ultimately enhancing metastatic traits. Deciphering the intricate interplay between ECM properties and organelle adaptations not only advances our understanding of cytoskeletal dynamics but also holds promise for the development of innovative anti-metastatic therapeutic strategies.

Retrospective analysis of epidemiologic features and clinical course of COVID-19 patients and comparison between vaccinated and unvaccinated patients.

At our Pneumology Department, we dealt with three waves of COVID-19 pandemics. The purpose of this study is to compare patients' epidemiological and clinical characteristics across waves and to assess the effect of vaccination on clinical presentation, course, and prognosis. From March 2020 to March 2022, a retrospective cohort study was conducted to compare patient characteristics. Based on the time of hospital admission, data from 456 patients were collected and divided into three groups (IW, IIW, and IIIW). In addition, we looked at the link between vaccination and clinical presentation and hospitalization outcome. The average age and comorbidities of patients increased, as did the worsening of respiratory conditions at admission (PaO2/FiO2 median 207 in IW, 95.5 in IIW, and 99 in IIIW). Continuous positive airway pressure (CPAP) was the primary respiratory support during the first wave, but an increase in the use of high flow nasal cannula and noninvasive ventilation was later observed, resulting in a higher hospital discharge rate and a lower intubation rate. Vaccinated patients had less severe COVID-19-related respiratory failure, a better clinical course, and a higher hospital discharge rate (71.4% in V-group vs 44.7% in NV-group, p<0.001). Patients' characteristics changed over the three waves, possibly due to virus mutations. The advancement of clinical and therapeutic management knowledge has contributed to a reduction in the severity of respiratory failure. The vaccination campaign improved the clinical course and reduced mortality.

AMPK is a mechano-metabolic sensor linking cell adhesion and mitochondrial dynamics to Myosin-dependent cell migration.

Cell migration is crucial for cancer dissemination. We find that AMP-activated protein kinase (AMPK) controls cell migration by acting as an adhesion sensing molecular hub. In 3-dimensional matrices, fast-migrating amoeboid cancer cells exert low adhesion/low traction linked to low ATP/AMP, leading to AMPK activation. In turn, AMPK plays a dual role controlling mitochondrial dynamics and cytoskeletal remodelling. High AMPK activity in low adhering migratory cells, induces mitochondrial fission, resulting in lower oxidative phosphorylation and lower mitochondrial ATP. Concurrently, AMPK inactivates Myosin Phosphatase, increasing Myosin II-dependent amoeboid migration. Reducing adhesion or mitochondrial fusion or activating AMPK induces efficient rounded-amoeboid migration. AMPK inhibition suppresses metastatic potential of amoeboid cancer cells in vivo, while a mitochondrial/AMPK-driven switch is observed in regions of human tumours where amoeboid cells are disseminating. We unveil how mitochondrial dynamics control cell migration and suggest that AMPK is a mechano-metabolic sensor linking energetics and the cytoskeleton.

Metabolic rewiring in MYC-driven medulloblastoma by BET-bromodomain inhibition.

Medulloblastoma (MB) is the most common malignant brain tumour in children. High-risk MB patients harbouring MYC amplification or overexpression exhibit a very poor prognosis. Aberrant activation of MYC markedly reprograms cell metabolism to sustain tumorigenesis, yet how metabolism is dysregulated in MYC-driven MB is not well understood. Growing evidence unveiled the potential of BET-bromodomain inhibitors (BETis) as next generation agents for treating MYC-driven MB, but whether and how BETis may affect tumour cell metabolism to exert their anticancer activities remains unknown. In this study, we explore the metabolic features characterising MYC-driven MB and examine how these are altered by BET-bromodomain inhibition. To this end, we employed an NMR-based metabolomics approach applied to the MYC-driven MB D283 and D458 cell lines before and after the treatment with the BETi OTX-015. We found that OTX-015 triggers a metabolic shift in both cell lines resulting in increased levels of myo-inositol, glycerophosphocholine, UDP-N-acetylglucosamine, glycine, serine, pantothenate and phosphocholine. Moreover, we show that OTX-015 alters ascorbate and aldarate metabolism, inositol phosphate metabolism, phosphatidylinositol signalling system, glycerophospholipid metabolism, ether lipid metabolism, aminoacyl-tRNA biosynthesis, and glycine, serine and threonine metabolism pathways in both cell lines. These insights provide a metabolic characterisation of MYC-driven childhood MB cell lines, which could pave the way for the discovery of novel druggable pathways. Importantly, these findings will also contribute to understand the downstream effects of BETis on MYC-driven MB, potentially aiding the development of new therapeutic strategies to combat medulloblastoma.

Amoeboid migration in health and disease: Immune responses versus cancer dissemination.

Cell migration is crucial for efficient immune responses and is aberrantly used by cancer cells during metastatic dissemination. Amoeboid migrating cells use myosin II-powered blebs to propel themselves, and change morphology and direction. Immune cells use amoeboid strategies to respond rapidly to infection or tissue damage, which require quick passage through several barriers, including blood, lymph and interstitial tissues, with complex and varied environments. Amoeboid migration is also used by metastatic cancer cells to aid their migration, dissemination and survival, whereby key mechanisms are hijacked from professionally motile immune cells. We explore important parallels observed between amoeboid immune and cancer cells. We also consider key distinctions that separate the lifespan, state and fate of these cell types as they migrate and/or fulfil their function. Finally, we reflect on unexplored areas of research that would enhance our understanding of how tumour cells use immune cell strategies during metastasis, and how to target these processes.

The amoeboid state as part of the epithelial-to-mesenchymal transition programme.

Cell migration is essential for many biological processes, while abnormal cell migration is characteristic of cancer cells. Epithelial cells become motile by undergoing epithelial-to-mesenchymal transition (EMT), and mesenchymal cells increase migration speed by adopting amoeboid features. This review highlights how amoeboid behaviour is not merely a migration mode but rather a cellular state - within the EMT spectra - by which cancer cells survive, invade and colonise challenging microenvironments. Molecular biomarkers and physicochemical triggers associated with amoeboid behaviour are discussed, including an amoeboid associated tumour microenvironment. We reflect on how amoeboid characteristics support metastasis and how their liabilities could turn into therapeutic opportunities.

Phytochemical Characterization of Olea europaea L. Cultivars of Cilento National Park (South Italy) through NMR-Based Metabolomics.

Olea europaea germplasm is constituted by a huge number of cultivars, each one characterized by specific features. In this context, endemic cultivars evolved for a very long period in a precise local area, developing very specific traits. These characteristics include the production and accumulation of phytochemicals, many of which are also responsible for the nutraceutical value of the drupes and of the oils therefrom. With the aim of obtaining information on the phytochemical profile of drupes of autochthonous cultivars of Cilento, Vallo di Diano and Alburni National Park, a metabolomics-based study was carried out on 19 selected cultivars. Multivariate data analysis of 1H-NMR data and 2D NMR analyses allowed the rapid identification of metabolites that were qualitatively and/or quantitatively varying among the cultivars. This study allowed to identify the cultivars Racioppella, Guglia, Pizzulella, Oliva amara, and Racioppa as the richest in health-promoting phenolic compounds. Furthermore, it showed a significant variability among the different cultivars, suggesting the possibility of using metabolic fingerprinting approaches for cultivar differentiation, once that further studies aimed at assessing the influence of growing conditions and environmental factors on the chemical profiles of single cultivars are carried out.

NMR Profiling of Ononis diffusa Identifies Cytotoxic Compounds against Cetuximab-Resistant Colon Cancer Cell Lines.

In the search of new natural products to be explored as possible anticancer drugs, two plant species, namely Ononis diffusa and Ononis variegata, were screened against colorectal cancer cell lines. The cytotoxic activity of the crude extracts was tested on a panel of colon cancer cell models including cetuximab-sensitive (Caco-2, GEO, SW48), intrinsic (HT-29 and HCT-116), and acquired (GEO-CR, SW48-CR) cetuximab-resistant cell lines. Ononis diffusa showed remarkable cytotoxic activity, especially on the cetuximab-resistant cell lines. The active extract composition was determined by NMR analysis. Given its complexity, a partial purification was then carried out. The fractions obtained were again tested for their biological activity and their metabolite content was determined by 1D and 2D NMR analysis. The study led to the identification of a fraction enriched in oxylipins that showed a 92% growth inhibition of the HT-29 cell line at a concentration of 50 µg/mL.

NMR-Based Plant Metabolomics in Nutraceutical Research: An Overview.

Few topics are able to channel the interest of researchers, the public, and industries, like nutraceuticals. The ever-increasing demand of new compounds or new sources of known active compounds, along with the need of a better knowledge about their effectiveness, mode of action, safety, etc., led to a significant effort towards the development of analytical approaches able to answer the many questions related to this topic. Therefore, the application of cutting edges approaches to this area has been observed. Among these approaches, metabolomics is a key player. Herewith, the applications of NMR-based metabolomics to nutraceutical research are discussed: after a brief overview of the analytical workflow, the use of NMR-based metabolomics to the search for new compounds or new sources of known nutraceuticals are reviewed. Then, possible applications for quality control and nutraceutical optimization are suggested. Finally, the use of NMR-based metabolomics to study the impact of nutraceuticals on human metabolism is discussed.

Lymphocytes exposed to vegetables grown in waters contaminated by anticancer drugs: metabolome alterations and genotoxic risks for human health.

Wastewater irrigation of crops may be effective to avoid depletion (about 70%) of freshwater resources. However, the use of reclaimed waters containing persistent microcontaminants such as antineoplastic drugs is of high environmental concern. These active compounds may affect human health with potentially severe adverse effects. To better understand the impact on human health following irrigation of crops with reused contaminated waters, we exposed four edible plants, Brassica rapa, Lactuca sativa, Raphanus sativus, and Triticum durum, to two commonly used antitumoral drugs: 5-fluorouracil (5-FU), and Cisplatin (CDDP), using metabolomics as a potential functional genomics tool to combine with genotoxicity experiments. The metabolome of the treated and untreated plants was analysed to detect biochemical alterations associated to the exposure, and the potential genotoxic damage related to human exposure to the treated plants was evaluated using the comet assay in human lymphocytes, which are characterized by high sensitivity to genotoxic substances. The edible species were able to assimilate 5-FU and CDDP during the treatment, affecting the biochemical pathways of these plants with subsequent metabolome modifications. These metabolic alterations differed according to the specific species used for the test. Furthermore, all vegetables treated with two concentrations of the selected drugs (10 and 100 µg/L) caused significant (p < 0.0001) genotoxic damage in the cells of the immune system at a higher level than in the lymphocytes directly exposed to single antineoplastic drugs.

NMR-based metabolomics and bioassays to study phytotoxic extracts and putative phytotoxins from Mediterranean plant species.

INTRODUCTION: Mediterranean plants are characterised by a high content of bioactive secondary metabolites that play important roles in plant-plant interactions as plant growth regulators and could be useful for the development of new eco-friendly herbicides. OBJECTIVE: An NMR-based metabolomics approach was reported to seek selective phytotoxic plant extracts and putative plant-derived active molecules. METHODS: Plant extracts derived from five Mediterranean donor species (Pistacia lentiscus, Bellis sylvestris, Phleum subulatum, Petrohrhagia saxifraga and Melilotus neapolitana) were used to treat the hydroponic cultures of three receiving plants (Triticum durum, Triticum ovatum and Avena fatua). Morphological analyses of the treated receiving plants were carried out. NMR-based metabolomics was applied both to characterise the donor plant extracts and to study the effects of the treatments on the receiving plants. RESULTS: This study allowed the identification of Melilotus neapolitana and Bellis sylvestris as phytotoxic plant and good candidates for further studies. Specifically, the NMR-based metabolomics investigation showed that these species affect a specific set of metabolites (such as sugars, amino and organic acids) and therefore metabolic pathways [i.e. tricarboxylic acid (TCA) cycle, amino acid metabolism, etc.] that are crucial for the plant growth and development. Moreover, it was possible to identify the metabolite(s) probably responsible for the phytotoxicity of the active extracts. CONCLUSION: The NMR-based metabolomics approach employed in this study led to the identification of two phytotoxic plant extracts and their putative active principles. These new insights will be of paramount importance in the future to find plant derived molecules endowed with phytotoxic activities.

Spectroscopic Characterization and Cytotoxicity Assessment towards Human Colon Cancer Cell Lines of Acylated Cycloartane Glycosides from Astragalus boeticus L.

In several European countries, especially in Sweden, the seeds of the species Astragalus boeticus L. were widely used as coffee substitutes during the 19th century. Nonetheless, data regarding the phytochemistry and the pharmacological properties of this species are currently extremely limited. Conversely, other species belonging to the Astragalus genus have already been extensively investigated, as they were used for millennia for treating various diseases, including cancer. The current work was addressed to characterize cycloartane glycosides from A. boeticus, and to evaluate their cytotoxicity towards human colorectal cancer (CRC) cell lines. The isolation of the metabolites was performed by using different chromatographic techniques, while their chemical structures were elucidated by nuclear magnetic resonance (NMR) (1D and 2D techniques) and electrospray-ionization quadrupole time-of-flight (ESI-QTOF) mass spectrometry. The cytotoxic assessment was performed in vitro by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays in Caco-2, HT-29 and HCT-116 CRC cells. As a result, the targeted phytochemical study of A. boeticus enabled the isolation of three new cycloartane glycosides, 6-O-acetyl-3-O-(4-O-malonyl)-ß-d-xylopyranosylcycloastragenol (1), 3-O-(4-O-malonyl)-ß-d-xylopyranosylcycloastragenol (2), 6-O-acetyl-25-O-ß-d-glucopyranosyl-3-O-ß-d-xylopyranosylcycloastragenol (3) along with two known compounds, 6-O-acetyl-3-O-ß-d-xylopyranosylcycloastragenol (4) and 3-O-ß-d-xylopyranosylcycloastragenol (5). Importantly, this work demonstrated that the acetylated cycloartane glycosides 1 and 4 might preferentially inhibit cell growth in the CRC cell model resistant to epidermal growth factor receptor (EGFR) inhibitors.

Urtica dioica L. inhibits proliferation and enhances cisplatin cytotoxicity in NSCLC cells via Endoplasmic Reticulum-stress mediated apoptosis.

Non-small cell lung cancer (NSCLC) is the most common type of lung cancer and the ineffectiveness of the current therapies seriously limits the survival rate of NSCLC patients. In the search for new antitumor agents, nature has played a pivotal role providing a variety of molecules, which are likely to exert selective anti-tumour properties. Herein, we investigated the antiproliferative potential of Urtica dioica L. extract (UD) against NSCLC cell models with low sensitivity to cisplatin, a cytotoxic agent largely employed to cure NSCLCs. UD inhibited cell proliferation in the selected cells, while no toxic effects were observed in normal lung cells. Furthermore, the co-treatment of UD and cisplatin notably sensitised NSCLC cells to cisplatin. Mechanistically, we discovered that UD-promoted endoplasmic reticulum (ER) stress via activation of the growth arrest and DNA damage-inducible gene 153 (GADD153) triggering apoptosis. We also performed an extensive NMR analysis of UD, identifying rutin and oxylipins as the main secondary metabolites present in the mixture. Additionally, we discovered that an oxylipins' enriched fraction contributes to the antiproliferative activity of the plant extract. In the future, this study may provide new chemical scaffolds for the design of anti-cancer agents that target NSCLCs with low sensitivity to cisplatinum.

Metabolomic approach for a rapid identification of natural products with cytotoxic activity against human colorectal cancer cells.

The discovery of bioactive compounds from natural sources entails an extremely lengthy process due to the timescale and complexity of traditional methodologies. In our study, we used a rapid NMR based metabolomic approach as tool to identify secondary metabolites with anti-proliferative activity against a panel of human colorectal cancer cell lines with different mutation profiles. For this purpose, fourteen Fabaceae species of Mediterranean vegetation were investigated using a double screening method: 1H NMR profiling enabled the identification of the main compounds present in the mixtures, whilst parallel biological assays allowed the selection of two plant extracts based on their strong anti-proliferative properties. Using high-resolution 2D NMR spectroscopy, putative active constituents were identified in the mixture and isolated by performing a bio-guided fractionation of the selected plant extracts. As a result, we found two active principles: a cycloartane glycoside and protodioscin derivative. Interestingly, these metabolites displayed a preferential anti-proliferative effect on colon cancer cell lines with an intrinsic resistance to anti-EGFR therapies. Our work provides an NMR-based metabolomic approach as a powerful and efficient tool to discover natural products with anticancer activities circumventing time-consuming procedures.

Prenylated phloroglucinols from Hypericum scruglii, an endemic species of Sardinia (Italy), as new dual HIV-1 inhibitors effective on HIV-1 replication.

In a search for new potential multitarget anti-HIV compounds from natural products, we have identified in Hypericum scruglii, an endemic and exclusive species of Sardinia (Italy), a potent plant lead. The phytochemical study of the hydroalcoholic extract obtained from its leaves led to the isolation of its most abundant secondary metabolites, belonging to different chemical classes. In particular, three phloroglucinols derivatives were identified, confirming their significance as chemotaxonomic markers of the Hypericum genus. Among them, the 3-(13-hydroxygeranyl)-1-(2'-methylbutanoyl)phloroglucinol was reported here for the first time. All six isolated compounds have been evaluated firstly for the inhibition of both Human Immunodeficiency Virus type 1 (HIV-1) Reverse Transcriptase (RT)-associated DNA Polymerase (RDDP) and Ribonuclease H (RNase H) activities, for the inhibition of HIV-1 integrase (IN) in biochemical assays, and also for their effect on viral replication. Among the isolated metabolites, three phloroglucinol derivatives and quercitrin were effective on both RT-associated RDDP and RNase H activities in biochemical assays. The same active compounds affected also HIV-1 IN strand transfer function, suggesting the involvement of the RNase H active site. Furthermore, phloroglucinols compounds, included the newly identified compound, were able to inhibit the HIV-1 replication in cell based assays.

2D-NMR investigation and in vitro evaluation of antioxidant, antigenotoxic and estrogenic/antiestrogenic activities of strawberry grape.

Strawberry grape is considered beneficial due to its extensive phytochemical properties. To expand the knowledge about the chemical constituents and the biological activities of the whole plant, 2D-NMR investigation has been carried out on pulp, peel, seeds, stalks and leaves. Catechin and epicatechin were identified as the main constituents of the seed extract, quercetin and ferulic acid were detected in the leaves and malvidin and cyanidin glucopyranoside in the peels. The leaf, stalk and seed extracts were found to be very rich in phytochemicals and were tested for their ability to reduce the mutagenicity and genotoxicity of standard agents via Salmonella mutagenicity assay and SOS chromotest, respectively. Moreover, the estrogen/antiestrogen-like activity was evaluated on the MCF-7 estrogen-responsive cells. Seed and stalk extracts had an elevated antimutagenic/antigenotoxic activity. Stalk extracts highly reduced the proliferative effect of natural estrogen, 17ß-estradiol.