bims-polyam Biomed News
on Polyamines
Issue of 2019–11–10
ten papers selected by
Sebastian J. Hofer, University of Graz and Alexander Ivanov, Engelhardt Institute of Molecular Biology



  1. Cell Death Dis. 2019 Nov 07. 10(11): 854
      Lung adenocarcinoma (LUAD) is one of the leading causes of cancer-related death worldwide. There is an urgent need to uncover the pathogenic mechanism to develop new treatments. Agmatinase (AGMAT) expression and its association with clinicopathological characteristics were analyzed via GEO, Oncomine, and TCGA databases, and IHC staining in human LUAD specimens. An EdU cell proliferation kit, propidiumiodide staining, colony formation, cell migration, and invasion assays, and a xenograft tumor model were used to detect the biological function of AGMAT in LUAD. Furthermore, the expression level of nitric oxide (NO) was detected using a DAF-FMDA fluorescent probe or nitrite assay kit, and further validated with Carboxy-PTIO (a NO scavenger). The roles of three isoforms of nitric oxide synthases (nNOS, eNOS, and iNOS) were validated using L-NAME (eNOS inhibitor), SMT (iNOS inhibitor), and spermidine (nNOS inhibitor). AGMAT expression was up-regulated in LUAD tissues. LUAD patients with high AGMAT levels were associated with poorer prognoses. AGMAT promoted LUAD tumorigenesis in NO released by iNOS both in vitro and in vivo. Importantly, NO signaling up-regulated the expression of cyclin D1 via activating the MAPK and PI3K/Akt-dependent c-myc activity, ultimately promoting the malignant proliferation of tumor cells. On the whole, AGMAT promoted NO release via up-regulating the expression of iNOS. High levels of NO drove LUAD tumorigenesis via activating MAPK and PI3K/Akt cascades. AGMAT might be a potential diagnostic and therapeutic target for LUAD patients.
    DOI:  https://doi.org/10.1038/s41419-019-2082-3
  2. Autophagy. 2019 Nov 06. 1-2
      Organismal aging is associated with compromised cellular function, which can be partially attributed to accumulation of cellular damage. Being the major, if not only, cellular bulk-degradation mechanism, macroautophagy (hereafter autophagy) declines with age in multiple tissues and organisms. Spermidine is an endogenous polyamine metabolite that also declines with age. It prolongs lifespan and improves tissue functions of model organisms in an autophagy-dependent manner. We report that autophagic flux is significantly reduced in B cells from old mice. Spermidine induces autophagy and improves the function of both old mouse and old human B cells. Mechanistically, spermidine post-translationally modifies (hypusinates) the translation factor EIF5A. Hypusinated EIF5A specifically regulates the synthesis of the master autophagy and lysosome transcription factor, TFEB (transcription factor EB). This pathway declines with age in both mice and humans, which may eventually lead to declining autophagy and impaired tissue functions in old individuals.
    Keywords:  Aging; B cells; EIF5A; TFEB; autophagy; hypusine; spermidine; translation
    DOI:  https://doi.org/10.1080/15548627.2019.1687967
  3. Nutrients. 2019 Nov 03. pii: E2637. [Epub ahead of print]11(11):
      N-carbamoyl putrescine (NCP), the decarboxylation derivative of citrulline, metabolically related to polyamines, may exert biological effects in mammals. The aim of this study was (i) to evaluate the nutritional properties of NCP in healthy rats and (ii) to determine the effect of NCP administration on muscle metabolism in malnourished old rats. The nutritional properties of NCP were first evaluated in 20 8-week-old male rats randomized to receive for two weeks a standard diet either alone (C group) or supplemented with NCP, 5 or 50 mg/kg/d. In a second study, 29 malnourished 18-month-old male rats were studied either before or after a 4-day refeeding with a standard diet either alone (REN group) or supplemented with NCP, 1 or 10 mg/kg/d. NCP had no effect on weight gain and body composition in either of the two studies. In healthy rats, muscle protein content was significantly increased in the soleus with NCP 5 mg/kg/d. A decrease in plasma glutamine and kidney spermine was observed at the 50 mg/kg/d dose; otherwise, no significant changes in plasma chemistry and tissue polyamines were observed. In malnutrition-induced sarcopenic old rats, refeeding with NCP 10 mg/kg/d was associated with higher tibialis weight and a trend for increased protein content in extensor digitorum longus (EDL). While the muscle protein synthesis rate was similar between groups, ribosomal protein S6 kinase was increased in tibialis and higher in the EDL in NCP-treated rats. The muscle RING-finger protein-1 expression was decreased in tibialis and urinary 3-methyl-histidine to creatinine ratio slightly lower with the supply of NCP. However, this initial period of refeeding was also associated with elevated fasted plasma triglycerides and glucose, significant in NCP groups, suggesting glucose intolerance and possibly insulin resistance. NCP was well-tolerated in healthy young-adults and in malnourished old rats. In healthy adults, NCP at 5 mg/kg/d induced a significant increase in protein content in the soleus, a type I fiber-rich muscle. In malnourished old rats, NCP supply during refeeding, may help to preserve lean mass by limiting protein breakdown; however, these effects may be limited in our model by a possible immediate refeeding-associated glucose intolerance.
    Keywords:  N-carbamoyl putrescine; catabolism; citrulline; malnutrition; muscle anabolism
    DOI:  https://doi.org/10.3390/nu11112637
  4. Metabolites. 2019 Nov 05. pii: E266. [Epub ahead of print]9(11):
      Yeast cells respond to heat stress by remodeling their gene expression, resulting in the changes of the corresponding proteins and metabolites. Compared to the intensively investigated transcriptome and proteome, the metabolic response to heat stress is not sufficiently characterized. Mitochondria have been recognized to play an essential role in heat stress tolerance. Given the compartmentalization of the cell, it is not clear if the heat stress-induced metabolic response occurs in mitochondria or in the cytosol. Therefore, a compartment-specific metabolite analysis was performed to analyze the heat stress-induced metabolic response in mitochondria and the cytoplasm. In this work, the isolated mitochondria and the cytoplasm of yeast cells grown at permissive temperature and cells adapting to heat stress were subjected to mass spectrometry-based metabolomics. Over a hundred metabolites could be identified, covering amino acid metabolism, energy metabolism, arginine metabolism, purine and pyrimidine metabolism, and others. Highly accumulated citrulline and reduced arginine suggested remodeled arginine metabolism. A stable isotope-labeled experiment was performed to analyze the heat stress-induced metabolic remodeling of the arginine metabolism, identifying activated de novo ornithine biosynthesis to support arginine and spermidine synthesis. The short-term increased spermidine and trehalose suggest their important roles as heat stress markers. These data provide metabolic clues of heat stress-induced metabolic remodeling, which helps in understanding the heat stress response.
    Keywords:  arginine metabolism; heat stress; metabolic remodeling; metabolomics; mitochondria; yeast
    DOI:  https://doi.org/10.3390/metabo9110266
  5. Ecotoxicol Environ Saf. 2019 Nov 04. pii: S0147-6513(19)31068-1. [Epub ahead of print] 109737
      The current manuscript presents the first report on the ameliorative roles of exogenous spermine (Spm) during prolonged fluoride-induced toxicity and oxidative damages in the susceptible rice cultivar, IR-64. The application of Spm increased the overall growth in the stressed seedlings by significantly restricting fluoride bioaccumulation within the shoots and roots. The Spm-treated stressed seedlings exhibited low chlorosis and induced activity of pyruvate dehydrogenase and nitrate reductase due to reduced accumulation and localization of reactive oxygen species (ROS) in the shoot and root. Spm-supplementation during stress reduced the levels of molecular damages by lowering malondialdehyde, electrolyte leakage and protein carbonylation, and lipoxygenase and protease activity due to effective detoxification of ROS by the antioxidants like proline, glycine-betaine, anthocyanin, flavonoids, phenolics and higher polyamines like Spm and spermidine. Excessive accumulation of the toxic methylglyoxal was reversed due to the activation of the glyoxalase system (comprising of glyoxalase I and II) and the ascorbate-glutathione cycle. Exogenous Spm also triggered the activity of superoxide dismutase, guaiacol peroxidase, glutathione peroxidase and phenylalanine ammonia lyase, which efficiently scavenged ROS in the stressed seedlings. Overall, Spm treatment mitigated the fluoride-induced injuries in IR-64 by reducing fluoride bioaccumulation and elaborately refining the various defence machineries.
    Keywords:  Antioxidants; Fluoride toxicity; Glyoxalase system; Reactive oxygen species; Rice; Spermine
    DOI:  https://doi.org/10.1016/j.ecoenv.2019.109737
  6. Sci Rep. 2019 Nov 05. 9(1): 16025
      The effect of foliar application of polyamines on roses (Rosa hybrida cv. 'Herbert Stevens') was investigated in a factorial experiment based on a completely randomized design with three replications in a greenhouse. Two factors were applied including polyamine type (putrescine, spermidine, and spermine) and polyamine concentration (0, 1, 2 and 4 mM). The recorded traits included root fresh and dry weight, root length, number of flowers, flower longevity, chlorophyll content, carotenoids, antioxidant enzymes activity (catalase, ascorbate peroxidase and guaiacol peroxidase) and some macronutrients such as nitrogen, phosphorus and potassium. The results showed that among polyamines, putrescine had the greatest effect on root dry weight; spermidine showed the greatest effect on root length, chlorophyll content, plant phosphorus and spermine affected root fresh weight and flower longevity most strongly. Polyamine concentration of 1 mM had the strongest effect on flower longevity, carotenoids, nitrogen and phosphorus content. The highest potassium rate was observed in treatments with the concentration of 4 mM. Polyamine treatments had no significant effect on the number of flowers per plant and antioxidant enzymes.
    DOI:  https://doi.org/10.1038/s41598-019-52547-1
  7. Chembiochem. 2019 Nov 08.
      Currently significant efforts are devoted to designing small molecules able to bind selectively to guanine-quadruplexes (G4s). These non-canonical DNA structures are implicated in various important biological processes and have been identified as potential targets for drug development. Previously, we reported a series of triphenylamine(TPA)-based compounds including macrocyclic polyamines, which display high affinity towards G4 DNA. Following from this initial work, herein we present a series of second-generation compounds, in which the central TPA has been functionalised with flexible and adaptive linear polyamines, aiming to maximise the selectivity towards G4 DNA. The acid-base properties of the new derivatives have been studied by means of potentiometric titrations, UV-Vis and fluorescence emission spectroscopies. The interaction with G4s and duplex DNA has been explored using FRET melting assays, fluorescence spectroscopy and circular dichroism. Compared to our previously TPA derivatives with macrocyclic substituents, the new ligands reported herein retain the G4 affinity, but display two orders of magnitude higher selectivity for G4 vs. duplex DNA, most likely due to the ability of the linear substituents to embrace the G4 structure.
    Keywords:  * G-quadruplex DNA * G4 selectivity * polyamine-based ligand *fluorescence
    DOI:  https://doi.org/10.1002/cbic.201900678
  8. ACS Omega. 2019 Oct 29. 4(18): 17850-17856
      The surface modification of nanoparticles (NPs) can enhance cellular and intracellular targeting. A new type of polyamine-modified gold NPs (AuNPs) are designed and synthesized, which can be selectively absorbed onto the cell membrane. AuNPs with an average diameter of 4.0 nm were prepared and modified with polyamine (R-4C) through amidation. In order to detect the distribution of NPs within cells by fluorescence imaging, AuNP@MPA-R-4C was functionalized with fluorescein isothiocyanate (FITC). The fluorescence-labled NPs AuNP@MPA-R-4C-FITC demonstrated minimal cytotoxicity in several cell lines. Both confocal laser scanning microscopy and transmission electron microscopy demonstrated that AuNP@MPA-R-4C-FITC was distributed on the cell membrane. Compared with the free organic dye, the modified AuNPs showed significantly increased accumulation on the cell membrane after treatment for only 10 min. These results suggested that AuNP@MPA-R-4C-FITC can be used as a bioprobe targeting the cell membrane for various biological applications.
    DOI:  https://doi.org/10.1021/acsomega.9b02579
  9. ACS Appl Mater Interfaces. 2019 Nov 04.
      A graphene aerogel (GA) with a three-dimensional (3D) structure, ultra-lightweight nature, and high hydrophobicity was simply fabricated by the one-step pyrolysis of glucose and ammonium chloride. The as-synthesized GA exhibited a 3D interconnected microporous architecture with a high surface area of ~2860 m2∙g-1 and pore volume of 2.24 cm3∙g-1. The hydrophobic GA (10 mg∙100 mL-1) demonstrated rapid and excellent adsorption performance for the removal of food toxins such as various biogenic amines (histamine, cadaverine and spermine) and the hazardous bacterium Staphylococcus aureus (a food contaminant and a cause of poor wound healing) from a liquid matrix with a maximum simultaneous adsorption capacity for multiple biogenic amines of > 85.19% (histamine), 74.1 % (cadaverine), and 70.11% (spermidine) and a 100% reduction in the viable cell count of S. aureus within 80 min of interaction. The outstanding adsorption capacity can be attributed to a highly interconnected porous network in the 3D architecture and a high surface-to-volume ratio. A case study using soy sauce spiked with multiple biogenic amines showed successful removal of toxins with excellent recyclability without any loss in absorption performance. Biocompatibility of the GA in terms of cell viability was observed even at high concentrations (83.46% and 75.28% at 25 and 50 mg.mL-1, respectively). Confirmatory biocompatibility testing was conducted via live/dead cell evaluation and the morphology of normal lung epithelial cells examined via scanning electron microscopy showed no cellular shrinkage. Moreover, GA showed excellent removal of live colonies of S. aureus from the food matrix and immunoblotting analysis showed elevated protein expression levels of β-catenin and α-SMA (α-smooth muscle actin). The biocompatible sugar-based GA could simultaneous adsorb multiple biogenic amines and live bacteria and was easy to regenerate via simple separation due to its high floatability, hydrophobicity, surface area, and porosity without any structural and functional loss, making it especially relevant for food safety and biomedical applications.
    DOI:  https://doi.org/10.1021/acsami.9b16989
  10. J Mol Biol. 2019 Nov 01. pii: S0022-2836(19)30622-9. [Epub ahead of print]
      Multidrug-resistant Gram-negative bacteria infection is particularly severe within the designated ESKAPE pathogens (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter species), which underscores the urgent need to explore alternative therapeutic strategies. The type III secretion system (T3SS) is considered to be a key virulence factor in many Gram-negative bacteria, and T3SS is in turn regulated by SpuE in P. aeruginosa, which is a spermidine binding protein from an ATP-binding cassette transporter family and highly conserved within ESKAPE pathogens. Here, we identified a potent anti-SpuE antagonistic antibody that allosterically inhibits the expression of T3SS and attenuates virulence of P. aeruginosa. X-ray crystallography and molecular dynamics simulations revealed that binding of antibody to SpuE induces a change in the dynamics of SpuE, which in turn may reduce spermidine uptake by P. aeruginosa. The antibody could serve as a template for developing novel biologics to target a broad spectrum of Gram-negative bacteria.
    Keywords:  SpuE; antibody phage display; multi-drug resistant Pseudomonas aeruginosa; spermidine importer; type III secretion system
    DOI:  https://doi.org/10.1016/j.jmb.2019.10.026