bims-polyam 2020-12-20 papers

bims-polyam

Biomed News

on Polyamines

Issue of 2020‒12‒20
ten papers selected by
Sebastian J. Hofer
University of Graz

ATP13A3 is a major component of the enigmatic mammalian polyamine transport system.
Spermidine attenuates bleomycin-induced lung fibrosis by inducing autophagy and inhibiting endoplasmic reticulum stress (ERS)-induced cell death in mice.
Exploring the structural basis of conformational alterations of myoglobin in the presence of spermine through computational modeling, molecular dynamics simulations, and spectroscopy methods.
Autophagy in T cells from aged donors is maintained by spermidine and correlates with function and vaccine responses.
Hypusination Orchestrates the Antimicrobial Response of Macrophages.
Hippocampal ornithine decarboxylase/spermidine pathway mediates H2S-alleviated cognitive impairment in diabetic rats: Involving enhancment of hippocampal autophagic flux.
De novo synthesis and salvage pathway coordinately regulate polyamine homeostasis and determine T cell proliferation and function.
A structurally unique Fusobacterium nucleatum tannase provides detoxicant activity against gallotannins and pathogen resistance.
Metabolomic Profiling of Drought-Tolerant and Susceptible Peanut (Arachis hypogaea L.) Genotypes in Response to Drought Stress.
Knockdown of Parkinson's disease-related gene ATP13A2 reduces tumorigenesis via blocking autophagic flux in colon cancer.

J Biol Chem. 2020 Dec 11. pii: jbc.RA120.013908. [Epub ahead of print]

ATP13A3 is a major component of the enigmatic mammalian polyamine transport system.

Norin Nabil Hamouda, Chris Van den Haute, Roeland Vanhoutte, Ragna Sannerud, Mujahid Azfar, Rupert Mayer, Álvaro Cortés Calabuig, Johannes V Swinnen, Patrizia Agostinis, Veerle Baekelandt, Wim Annaert, Francis Impens, Steven H L Verhelst, Jan Eggermont, Shaun Martin, Peter Vangheluwe.

  Polyamines, such as putrescine, spermidine and spermine, are physiologically important polycations, but the transporters responsible for their uptake in mammalian cells remain poorly characterized. Here, we reveal a new component of the mammalian polyamine transport system (PTS) using CHO-MG cells, a widely used model to study alternative polyamine uptake routes and characterize polyamine transport inhibitors for therapy. CHO-MG cells present polyamine uptake deficiency and resistance to a toxic polyamine biosynthesis inhibitor MGBG (methylglyoxal bis- (guanylhydrazone)), but the molecular defects responsible for these cellular characteristics remain unknown. By genome sequencing of CHO-MG cells, we identified mutations in an unexplored gene, ATP13A3, and found disturbed mRNA and protein expression. ATP13A3 encodes for an orphan P5B-ATPase (ATP13A3), a P-type transport ATPase that represents a candidate polyamine transporter. Interestingly, ATP13A3 complemented the putrescine transport deficiency and MGBG resistance of CHO-MG cells, whereas its knockdown in wild-type cells induced a CHO-MG phenotype, demonstrating a decrease in putrescine uptake and MGBG sensitivity. Taken together, our findings identify ATP13A3 as a major component of the mammalian PTS that confers sensitivity to MGBG and that has been previously genetically linked with pulmonary arterial hypertension.

Keywords:  ATP13A2; ATPase; P-type ATPase; P5B-ATPase; endosome; polyamine; polyamine transport system; putrescine; transport; transporter

DOI:  https://doi.org/10.1074/jbc.RA120.013908
Exp Mol Med. 2020 Dec 14.

Spermidine attenuates bleomycin-induced lung fibrosis by inducing autophagy and inhibiting endoplasmic reticulum stress (ERS)-induced cell death in mice.

Ae Rin Baek, Jisu Hong, Ki Sung Song, An Soo Jang, Do Jin Kim, Su Sie Chin, Sung Woo Park.

Spermidine is an endogenous biological polyamine that plays various longevity-extending roles and exerts antioxidative, antiaging, and cell growth-promoting effects. We previously reported that spermidine levels were significantly reduced in idiopathic pulmonary fibrosis (IPF) of the lung. The present study assessed the potential beneficial effects of spermidine on lung fibrosis and investigated the possible mechanism. Lung fibrosis was established in mice using bleomycin (BLM), and exogenous spermidine was administered daily by intraperitoneal injection (50 mg/kg in phosphate-buffered saline). BLM-induced alveolar epithelial cells showed significant increases in apoptosis and endoplasmic reticulum stress (ERS)-related mediators, and spermidine attenuated BLM-induced apoptosis and activation of the ERS-related pathway. Senescence-associated β-gal staining and decreased expression of p16 and p21 showed that spermidine ameliorated BLM-induced premature cellular senescence. In addition, spermidine enhanced beclin-1-dependent autophagy and autophagy modulators in IPF fibroblasts and BLM-induced mouse lungs, in which inflammation and collagen deposition were significantly decreased. This beneficial effect was related to the antiapoptotic downregulation of the ERS pathway, antisenescence effects, and autophagy activation. Our findings suggest that spermidine could be a therapeutic agent for IPF treatment.

DOI: https://doi.org/10.1038/s12276-020-00545-z
J Biomol Struct Dyn. 2020 Dec 14. 1-14

Exploring the structural basis of conformational alterations of myoglobin in the presence of spermine through computational modeling, molecular dynamics simulations, and spectroscopy methods.

Rasoul Eslami-Farsani, Sadegh Farhadian, Behzad Shareghi.

  Spermine as polyamines can have interaction with the myoglobin (Mb). The intent of this pondering to evaluate the impact of spermine on Mb properties, for example, the structure and thermal stability. For this analysis, the following approaches are employed. Thermodynamics, molecular dynamics (MD), and docking and the use of other spectroscopic procedures. The results of fluorescence spectroscopy and docking showed that binding spermine to Mb was spontaneous. Spermine quenched the fluorescence of Mb through the static quenching process. The thermal stability of Mb was incremented when the concentration of spermine increased. The CD spectra showed Mb's secondary structure shift with a rise in β-sheet and a decrease in α-helicity Mb's in spermine presence. Molecular docking and MD simulation outcomes demonstrate that electrostatic forces show a critical function in stabilizing of this complex, which is in conforming to spectroscopic results. Communicated by Ramaswamy H. Sarma.

Keywords:  MD simulation; myoglobin; quenching mechanism; spermine; thermal stability

DOI:  https://doi.org/10.1080/07391102.2020.1848633
Elife. 2020 Dec 15. pii: e57950. [Epub ahead of print]9

Autophagy in T cells from aged donors is maintained by spermidine and correlates with function and vaccine responses.

Ghada Alsaleh, Isabel Panse, Leo Swadling, Hanlin Zhang, Felix Clemens Richter, Alain Meyer, Janet Lord, Eleanor Barnes, Paul Klenerman, Christopher Green, Anna Katharina Simon.

  Vaccines are powerful tools to develop immune memory to infectious diseases and prevent excess mortality. In older adults, however vaccines are generally less efficacious and the molecular mechanisms that underpin this remain largely unknown. Autophagy, a process known to prevent aging, is critical for the maintenance of immune memory in mice. Here, we show that autophagy is specifically induced in vaccine-induced antigen-specific CD8+ T cells in healthy human volunteers. In addition, reduced IFNγ secretion by RSV-induced T cells in older vaccinees correlates with low autophagy levels. We demonstrate that levels of the endogenous autophagy-inducing metabolite spermidine fall in human T cells with age. Spermidine supplementation in T cells from old donors recovers their autophagy level and function, similar to young donors' cells, in which spermidine biosynthesis has been inhibited. Finally, our data show that endogenous spermidine maintains autophagy via the translation factor eIF5A and transcription factor TFEB. In summary, we have provided evidence for the importance of autophagy in vaccine immunogenicity in older humans and uncovered two novel drug targets that may increase vaccination efficiency in the aging context.

Keywords:  TFEB; autophagy; human; human T cells; immunology; inflammation; spermidine; vaccine

DOI:  https://doi.org/10.7554/eLife.57950
Cell Rep. 2020 Dec 15. pii: S2211-1247(20)31499-6. [Epub ahead of print]33(11): 108510

Hypusination Orchestrates the Antimicrobial Response of Macrophages.

Alain P Gobert, Jordan L Finley, Yvonne L Latour, Mohammad Asim, Thaddeus M Smith, Thomas G Verriere, Daniel P Barry, Margaret M Allaman, Alberto G Delagado, Kristie L Rose, M Wade Calcutt, Kevin L Schey, Johanna C Sierra, M Blanca Piazuelo, Raghavendra G Mirmira, Keith T Wilson.

  Innate responses of myeloid cells defend against pathogenic bacteria via inducible effectors. Deoxyhypusine synthase (DHPS) catalyzes the transfer of the N-moiety of spermidine to the lysine-50 residue of eukaryotic translation initiation factor 5A (EIF5A) to form the amino acid hypusine. Hypusinated EIF5A (EIF5AHyp) transports specific mRNAs to ribosomes for translation. We show that DHPS is induced in macrophages by two gastrointestinal pathogens, Helicobacter pylori and Citrobacter rodentium, resulting in enhanced hypusination of EIF5A. EIF5AHyp was also increased in gastric macrophages from patients with H. pylori gastritis. Furthermore, we identify the bacteria-induced immune effectors regulated by hypusination. This set of proteins includes essential constituents of antimicrobial response and autophagy. Mice with myeloid cell-specific deletion of Dhps exhibit reduced EIF5AHyp in macrophages and increased bacterial burden and inflammation. Thus, regulation of translation through hypusination is a critical hallmark of the defense of eukaryotic hosts against pathogenic bacteria.

Keywords:  Citrobacter rodentium; Helicobacter pylori; bacterial infection; gastrointestinal tract; hypusine; innate immunity; macrophages; polyamines

DOI:  https://doi.org/10.1016/j.celrep.2020.108510
J Adv Res. 2021 Jan;27 31-40

Hippocampal ornithine decarboxylase/spermidine pathway mediates H2S-alleviated cognitive impairment in diabetic rats: Involving enhancment of hippocampal autophagic flux.

Xuan Kang, Cheng Li, Yan Xie, Ling-Li He, Fan Xiao, Ke-Bin Zhan, Yi-Yun Tang, Xiang Li, Xiao-Qing Tang.

  Introduction: We have previously demonstrated the antagonistic role of hydrogen sulfide (H2S) in the cognitive dysfunction of streptozotocin (STZ)-induced diabetic rats. It has been confirmed that the impaired hippocampal autophagic flux has a key role in the pathogenesis of cognitive impairment and that ornithine decarboxylase (ODC)/spermidine (Spd) pathway plays an important role in the formation of memory by promoting autophagic flux.Objectives: To investigate the roles of hippocampal ODC/Spd pathway and autophagic flux in H2S-attenuated cognitive impairment in STZ-induced diabetic rats.
Methods: Cognitive function is judged by the novel objective recognition task (NOR), the Y-maze, and the Morris water maze (MWM) tests. The ODC/Spd pathway in hippocampus was evaluated using the expression of ODC detected by western blot and the level of Spd assayed by GC-MS. Autophagic flux was assessed using the expressions of Beclin-1, LC3II/I, and P62 detected by western blot, and the number of autophagosomes observed by transmission electron microscope.
Results: Sodium hydrosulfide (NaHS, a donor of H2S) markedly improved the autophagic flux in the hippocampus of STZ-exposed rats, as evidenced by a decrease in the number of autophagosomes as wells as downregulations in the expressions of LC3-II, Beclin-1, and P62 in the hippocampus of cotreatment with NaHS and STZ rats. NaHS also up-regulated the expression of ODC and the level of Spd in the hippocampus of STZ-induced diabetic rats. Furthermore, inhibited hippocampal ODC/Spd pathway by difluoromethylornithine (DFMO) markedly reversed the protections of NaHS against the hippocampal autophagic flux impairment as well as the cognitive dysfunction in STZ-exposed rats.
Conclusion: These findings indicated that improving hippocampal autophagic flux plays a key role in H2S-attenuated cognitive impairment in STZ-induced diabetic rats, as results of up-regulating hippocampal ODC/Spd pathway.

Keywords:  Autophagic flux; Cognitive impairment; Diabetes; Hydrogen sulfide; Ornithine decarboxylase/spermidine pathway

DOI:  https://doi.org/10.1016/j.jare.2020.06.007
Sci Adv. 2020 Dec;pii: eabc4275. [Epub ahead of print]6(51):

De novo synthesis and salvage pathway coordinately regulate polyamine homeostasis and determine T cell proliferation and function.

Ruohan Wu, Xuyong Chen, Siwen Kang, Tingting Wang, Jn Rashida Gnanaprakasam, Yufeng Yao, Lingling Liu, Gaofeng Fan, Mark R Burns, Ruoning Wang.

Robust and effective T cell-mediated immune responses require proper allocation of metabolic resources through metabolic pathways to sustain the energetically costly immune response. As an essential class of polycationic metabolites ubiquitously present in all living organisms, the polyamine pool is tightly regulated by biosynthesis and salvage pathway. We demonstrated that arginine is a major carbon donor and glutamine is a minor carbon donor for polyamine biosynthesis in T cells. Accordingly, the dependence of T cells can be partially relieved by replenishing the polyamine pool. In response to the blockage of biosynthesis, T cells can rapidly restore the polyamine pool through a compensatory increase in extracellular polyamine uptake, indicating a layer of metabolic plasticity. Simultaneously blocking synthesis and uptake depletes the intracellular polyamine pool, inhibits T cell proliferation, and suppresses T cell inflammation, indicating the potential therapeutic value of targeting the polyamine pool for managing inflammatory and autoimmune diseases.

DOI: https://doi.org/10.1126/sciadv.abc4275
Microb Biotechnol. 2020 Dec 18.

A structurally unique Fusobacterium nucleatum tannase provides detoxicant activity against gallotannins and pathogen resistance.

José Miguel Mancheño, Estíbaliz Atondo, Julen Tomás-Cortázar, José Luís Lavín, Laura Plaza-Vinuesa, Itziar Martín-Ruiz, Diego Barriales, Ainhoa Palacios, Claudio Daniel Navo, Leticia Sampedro, Ainize Peña-Cearra, Miguel Ángel Pascual-Itoiz, Janire Castelo, Ana Carreras-González, Donatello Castellana, Aize Pellón, Susana Delgado, Patricia Ruas-Madiedo, Blanca de Las Rivas, Leticia Abecia, Rosario Muñoz, Gonzalo Jiménez-Osés, Juan Anguita, Héctor Rodríguez.

Colorectal cancer pathogenesis and progression is associated with the presence of Fusobacterium nucleatum and the reduction of acetylated derivatives of spermidine, as well as dietary components such as tannin-rich foods. We show that a new tannase orthologue of F. nucleatum (TanBFnn ) has significant structural differences with its Lactobacillus plantarum counterpart affecting the flap covering the active site and the accessibility of substrates. Crystallographic and molecular dynamics analysis revealed binding of polyamines to a small cavity that connects the active site with the bulk solvent which interact with catalytically indispensable residues. As a result, spermidine and its derivatives, particularly N8 -acetylated spermidine, inhibit the hydrolytic activity of TanBFnn and increase the toxicity of gallotannins to F. nucleatum. Our results support a model in which the balance between the detoxicant activity of TanBFnn and the presence of metabolic inhibitors can dictate either conducive or unfavourable conditions for the survival of F. nucleatum.

DOI: https://doi.org/10.1111/1751-7915.13732
ACS Omega. 2020 Dec 08. 5(48): 31209-31219

Metabolomic Profiling of Drought-Tolerant and Susceptible Peanut (Arachis hypogaea L.) Genotypes in Response to Drought Stress.

Srutiben A Gundaraniya, Padma S Ambalam, Rukam S Tomar.

Peanut is frequently constrained by extreme environmental conditions such as drought. To reveal the involvement of metabolites, TAG 24 (drought-tolerant) and JL 24 (drought-sensitive) peanut genotypes were investigated under control and 20% PEG 6000-mediated water scarcity conditions at the seedling stage. Samples were analyzed by gas chromatography-mass spectrometry (GC-MS) to identify untargeted metabolites and targeted metabolites, i.e., polyamines and polyphenols by high-performance liquid chromatography (HPLC) and ultrahigh-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS), respectively. The principal component analysis (PCA), partial least-squares discriminant analysis (PLS-DA), heat map, and cluster analysis were applied to the metabolomics data obtained by the GC-MS technique to determine the important metabolites for drought tolerance. Among 46 resulting metabolites, pentitol, phytol, xylonic acid, d-xylopyranose, stearic acid, and d-ribose were important drought-responsive metabolites. Agmatine and cadaverine were present in TAG 24 leaves and roots, respectively, during water-deficit conditions and believed to be the potential polyamines for drought tolerance. Polyphenols such as syringic acid and vanillic acid were produced more in the leaves of TAG 24, while catechin production was high in JL 24 during stress conditions. Seven metabolic pathways, namely, galactose metabolism, starch and sucrose metabolism, fructose and mannose metabolism, pentose and glucuronate interconversion, propanoate metabolism, amino sugar and nucleotide sugar metabolism, and biosynthesis of unsaturated fatty acids were significantly affected by water-deficit conditions. This study provides valuable information about the metabolic response of peanut to drought stress and metabolites identified, which encourages further study by transcriptome and proteomics to improve drought tolerance in peanut.

DOI: https://doi.org/10.1021/acsomega.0c04601
Cell Biosci. 2020 Dec 11. 10(1): 144

Knockdown of Parkinson's disease-related gene ATP13A2 reduces tumorigenesis via blocking autophagic flux in colon cancer.

Qian Chen, Li Zhong, Chao Zhou, Yan Feng, Quan-Xing Liu, Dong Zhou, Xiao Lu, Guang-Sheng Du, Dan Jian, Hao Luo, Dong Wang, Hong Zheng, Yuan Qiu.

  BACKGROUND: Accumulating evidence shows that Parkinson's disease is negatively associated with colon cancer risk, indicating that Parkinson's disease family proteins may be involved in the initiation of colon cancer. Here, we aimed to identify a Parkinson's disease-related gene involved in colon cancer, elucidate the underlying mechanisms, and test whether it can be used as a target for cancer therapy.METHODS: We first screened colon cancer and normal tissues for differential expression of Parkinson's disease-associated genes and identified ATP13A2, which encodes cation-transporting ATPase 13A2, as a putative marker for colon cancer. We next correlated ATP13A2 expression with colon cancer prognosis. We performed a series of ATP13A2 knockdown and overexpression studies in vitro to identify the contribution of ATP13A2 in the stemness and invasive capacity of colon cancer cells. Additionally, autophagy flux assay were determined to explore the mechanism of ATP13A2 induced stemness. Finally, we knocked down ATP13A2 in mice using siRNA to determine whether it can be used as target for colon cancer treatment.
RESULTS: Colon cancer patients with high ATP13A2 expression exhibit shorter overall survival than those with low ATP13A2. Functionally, ATP13A2 acts as a novel stimulator of stem-like traits. Furthermore, knockdown of ATP13A2 in HCT116 resulted in decreased levels of cellular autophagy. Additionally, bafilomycin A1, an autophagy inhibitor, reversed the ATP13A2-induced stemness of colon cancer cells. Lastly treatment with ATP13A2 siRNA reduced the volume of colon cancer xenografts in mice.
CONCLUSIONS: The PD-associated gene ATP13A2 is involved in colon cancer stemness through regulation of autophagy. Furthermore, ATP13A2 is a novel prognostic biomarker for colon cancer and is a potential target for colon cancer therapy.

Keywords:  ATP13A2; Autophagy; Colon cancer; Tumorigenesis

DOI:  https://doi.org/10.1186/s13578-020-00506-z