bims-polyam 2019-10-06 papers

on Polyamines

Issue of 2019–10–06
ten papers selected by
Sebastian J. Hofer, University of Graz and Alexander Ivanov, Engelhardt Institute of Molecular Biology

The complete loss of function of the SMS gene results in a severe form of Snyder-Robinson syndrome.
Polyamines and Aging: A CLEAR Connection?
Polyamine supplementation reduces DNA damage in adipose stem cells cultured in 3-D.
The Polyamine Spermidine Modulates the Production of the Bacterial Genotoxin Colibactin.
Fruit Architecture in Polyamine-Rich Tomato Germplasm Is Determined via a Medley of Cell Cycle, Cell Expansion, and Fruit Shape Genes.
Multi-Omics Studies Demonstrate Toxoplasma gondii-Induced Metabolic Reprogramming of Murine Dendritic Cells.
Altered metabolomic profile of dual-species biofilm: Interactions between Proteus mirabilis and Candida albicans.
Role of putrescine in ovary and embryo development in fruit bat Cynopterus sphinx during embryonic diapause.
Caspase recruitment domain family member 10 regulates carbamoyl phosphate synthase 1 and promotes cancer growth in bladder cancer cells.
Functional involvement of acid-sensing ion channel 3 in the swallowing reflex in rats.

Eur J Med Genet. 2019 Sep 30. pii: S1769-7212(19)30460-4. [Epub ahead of print] 103777

The complete loss of function of the SMS gene results in a severe form of Snyder-Robinson syndrome.

Lise Larcher, Joy W Norris, Elodie Lejeune, Julien Buratti, Cyril Mignot, Catherine Garel, Boris Keren, Charles E Schwartz, Sandra Whalen.

  Snyder-Robinson syndrome (SRS) is an X-linked syndromic intellectual disability condition caused by variants in the spermine synthase gene (SMS). The syndrome is characterized by facial dysmorphism, thin body build, kyphoscoliosis, osteoporosis, hypotonia, developmental delay and associated neurological features (seizures, unsteady gait, abnormal speech). Until now, only missense variants with a functionally characterized partial loss of function (LoF) have been described. Here we describe the first complete LoF variant, Met303Lysfs*, in a male patient with a severe form of Snyder-Robinson syndrome. He presented with multiple malformations and severly delayed development, and died at 4 months of age. Functional in vitro assays showed a complete absence of functional SMS protein. Taken together, our findings and those of previously reported patients confirm that pathogenic variants of SMS are indeed LoF and that there might exist a genotype-phenotype correlation between the type of variant and the severity of the syndrome.

Keywords:  Genotype-phenotype correlation; Loss of function; Multiple congenital anomalies syndrome; SMS; Snyder-robinson syndrome

DOI:  https://doi.org/10.1016/j.ejmg.2019.103777
Mol Cell. 2019 Oct 03. pii: S1097-2765(19)30688-4. [Epub ahead of print]76(1): 5-7

Polyamines and Aging: A CLEAR Connection?

Douglas R Green.

A decline in polyamine levels with age has been implicated in the pathophysiology of aging, and nutritional supplementation of spermidine can reduce age-related pathology and increase lifespan in a number of different organisms. In this issue of Molecular Cell, Zhang and colleagues provide a mechanistic link between polyamines, autophagy, and aging.

DOI: https://doi.org/10.1016/j.molcel.2019.09.003
Sci Rep. 2019 Oct 03. 9(1): 14269

Polyamine supplementation reduces DNA damage in adipose stem cells cultured in 3-D.

Manuela Minguzzi, Serena Guidotti, Daniela Platano, Stefania D'Adamo, Silvia Cetrullo, Elisa Assirelli, Spartaco Santi, Erminia Mariani, Giovanni Trisolino, Giuseppe Filardo, Flavio Flamigni, Rosa Maria Borzì.

According to previous research, natural polyamines exert a role in regulating cell committment and differentiation from stemness during skeletal development. In order to assess whether distinct polyamine patterns are associated with different skeletal cell types, primary cultures of stem cells, chondrocytes or osteoblasts were dedicated for HPLC analysis of intracellular polyamines. Spermine (SPM) and Spermidine (SPD) levels were higher in adipose derived stem cells (ASC) compared to mature skeletal cells, i.e. chondrocytes and osteoblasts, confirming the connection of polyamine content with stemness. To establish whether polyamines can protect ASC against oxidative DNA damage in a 3-D differentiation model, the level of γH2AX was measured by western blot, and found to correlate with age and BMI of patients. Addition of either polyamine to ASC was able to hinder DNA damage in the low micromolecular range, with marked reduction of γH2AX level at 10 µM SPM and 5 µM SPD. Molecular analysis of the mechanisms that might underlie the protective effect of polyamine supplementation evidences a possible involvement of autophagy. Altogether, these results support the idea that polyamines are able to manage both stem cell differentiation and cell oxidative damage, and therefore represent appealing tools for regenerative and cell based applications.

DOI: https://doi.org/10.1038/s41598-019-50543-z
mSphere. 2019 Oct 02. pii: e00414-19. [Epub ahead of print]4(5):

The Polyamine Spermidine Modulates the Production of the Bacterial Genotoxin Colibactin.

Camille V Chagneau, Christophe Garcie, Nadège Bossuet-Greif, Sophie Tronnet, Alexander O Brachmann, Jörn Piel, Jean-Philippe Nougayrède, Patricia Martin, Eric Oswald.

  Colibactin is a polyketide/nonribosomal peptide produced by Escherichia coli strains that harbor the pks island. This toxin induces DNA double-strand breaks and DNA interstrand cross-links in infected eukaryotic cells. Colibactin-producing strains are found associated with colorectal cancer biopsy specimens and promote intestinal tumor progression in various murine models. Polyamines are small polycationic molecules produced by both microorganisms and eukaryotic cells. Their levels are increased in malignancies, where they contribute to disease progression and metastasis. In this study, we demonstrated that the endogenous spermidine synthase SpeE is required for full genotoxic activity of colibactin-producing E. coli Supplying spermidine in a ΔspeE pks + E. coli strain restored genotoxic activity. Spermidine is involved in the autotoxicity linked to colibactin and is required for direct damaging activity on DNA. The production of the colibactin prodrug motif is impaired in ΔspeE mutants. Therefore, we demonstrated that spermidine has a direct impact on colibactin synthesis.IMPORTANCE Colibactin-producing Escherichia coli strains are associated with cancerous and precancerous colorectal tissues and are suspected of promoting colorectal carcinogenesis. In this study, we describe a new interplay between the synthesis of the genotoxin colibactin and the polyamine spermidine. Polyamines are highly abundant in cancer tissue and are associated with cell proliferation. The need for spermidine in genotoxic activity provides a new perspective on the role of these metabolites in the pathogenicity of colibactin-producing E. coli strains in colorectal cancer.

Keywords:  Escherichia coli ; biosynthesis; colorectal cancer; genotoxic colibactin; polyamines

DOI:  https://doi.org/10.1128/mSphere.00414-19
Plants (Basel). 2019 Sep 29. pii: E387. [Epub ahead of print]8(10):

Fruit Architecture in Polyamine-Rich Tomato Germplasm Is Determined via a Medley of Cell Cycle, Cell Expansion, and Fruit Shape Genes.

Raheel Anwar, Shazia Fatima, Autar K Mattoo, Avtar K Handa.

  Shape and size are important features of fruits. Studies using tomatoes expressing yeast Spermidine Synthase under either a constitutive or a fruit-ripening promoter showed obovoid fruit phenotype compared to spherical fruit in controls, suggesting that polyamines (PAs) have a role in fruit shape. The obovoid fruit pericarp exhibited decreased cell layers and pericarp thickness compared to wild-type fruit. Transgenic floral buds and ovaries accumulated higher levels of free PAs, with the bound form of PAs being predominant. Transcripts of the fruit shape genes, SUN1 and OVATE, and those of CDKB2, CYCB2, KRP1 and WEE1 genes increased significantly in the transgenic ovaries 2 and 5 days after pollination (DAP). The levels of cell expansion genes CCS52A/B increased at 10 and 20 DAP in the transgenic fruits and exhibited negative correlation with free or bound forms of PAs. In addition, the cell layers and pericarp thickness of the transgenic fruits were inversely associated with free or bound PAs in 10 and 20 DAP transgenic ovaries. Collectively, these results provide evidence for a linkage between PA homeostasis and expression patterns of fruit shape, cell division, and cell expansion genes during early fruit development, and suggest role(s) of PAs in tomato fruit architecture.

Keywords:  cell division; cell expansion; fruit shape; putrescine; spermidine; spermidine synthase; spermine; tomato

DOI:  https://doi.org/10.3390/plants8100387
Front Cell Infect Microbiol. 2019 ;9 309

Multi-Omics Studies Demonstrate Toxoplasma gondii-Induced Metabolic Reprogramming of Murine Dendritic Cells.

Kerrie E Hargrave, Stuart Woods, Owain Millington, Susan Chalmers, Gareth D Westrop, Craig W Roberts.

  Toxoplasma gondii is capable of actively invading almost any mammalian cell type including phagocytes. Early events in phagocytic cells such as dendritic cells are not only key to establishing parasite infection, but conversely play a pivotal role in initiating host immunity. It is now recognized that in addition to changes in canonical immune markers and mediators, alteration in metabolism occurs upon activation of phagocytic cells. These metabolic changes are important for supporting the developing immune response, but can affect the availability of nutrients for intracellular pathogens including T. gondii. However, the interaction of T. gondii with these cells and particularly how infection changes their metabolism has not been extensively investigated. Herein, we use a multi-omics approach comprising transcriptomics and metabolomics validated with functional assays to better understand early events in these cells following infection. Analysis of the transcriptome of T. gondii infected bone marrow derived dendritic cells (BMDCs) revealed significant alterations in transcripts associated with cellular metabolism, activation of T cells, inflammation mediated chemokine and cytokine signaling pathways. Multivariant analysis of metabolomic data sets acquired through non-targeted liquid chromatography mass spectroscopy (LCMS) identified metabolites associated with glycolysis, the TCA cycle, oxidative phosphorylation and arginine metabolism as major discriminants between control uninfected and T. gondii infected cells. Consistent with these observations, glucose uptake and lactate dehydrogenase activity were upregulated in T. gondii infected BMDC cultures compared with control BMDCs. Conversely, BMDC mitochondrial membrane potential was reduced in T. gondii-infected cells relative to mitochondria of control BMDCs. These changes to energy metabolism, similar to what has been described following LPS stimulation of BMDCs and macrophages are often termed the Warburg effect. This metabolic reprogramming of cells has been suggested to be an important adaption that provides energy and precursors to facilitate phagocytosis, antigen processing and cytokine production. Other changes to BMDC metabolism are evident following T. gondii infection and include upregulation of arginine degradation concomitant with increased arginase-1 activity and ornithine and proline production. As T. gondii is an arginine auxotroph the resultant reduced cellular arginine levels are likely to curtail parasite multiplication. These results highlight the complex interplay of BMDCs and parasite metabolism within the developing immune response and the consequences for adaptive immunity and pathogen clearance.

Keywords:  LPS stimulation; Toxoplasma gondii; Warburg effect; arginine metabolism; dendritic cells; immunometabolism; multi-omics

DOI:  https://doi.org/10.3389/fcimb.2019.00309
Microbiol Res. 2019 Sep 23. pii: S0944-5013(19)30598-1. [Epub ahead of print]230 126346

Altered metabolomic profile of dual-species biofilm: Interactions between Proteus mirabilis and Candida albicans.

Didem Kart, Samiye Yabanoglu Ciftci, Emirhan Nemutlu.

  In this study, we aimed to determine the interspecies interactions between Proteus mirabilis and Candida albicans. Mono and dual-species biofilms were grown in a microtiter plate and metabolomic analysis of the biofilms was performed. The effects of togetherness of two species on the expression levels of candidal virulence genes and urease and swarming activities of P.mirabilis were investigated. The growth of C.albicans was inhibited by P.mirabilis whereas the growth and swarming activity of P.mirabilis were increased by C.albicans. The inhibition of Candida cell growth was found to be biofilm specific. The alteration was not detected in urease activity. The expressions of EFG1, HWP1 and SAP2 genes were significantly down-regulated, however, LIP1 was upregulated by P.mirabilis. In the presence of P.mirabilis carbonhydrates, amino acids, polyamine and lipid metabolisms were altered in C.albicans. Interestingly, the putrescine level was increased up to 230 fold in dual-species biofilm compared to monospecies C.albicans biofilm. To our knowledge, this is the first study to investigate the impact of each microbial pathogen on the dual microbial environment by integration of metabolomic data.

Keywords:  Candida albicans; Dual-species biofilm; GC–MS; Metabolomic; Proteus mirabilis; Putrescine

DOI:  https://doi.org/10.1016/j.micres.2019.126346
Mol Reprod Dev. 2019 Oct 03.

Role of putrescine in ovary and embryo development in fruit bat Cynopterus sphinx during embryonic diapause.

Anuradha, Arnab Banerjee, Amitabh Krishna.

  The aim of this study was to evaluate the effect of putrescine on ovarian activity and the rate of embryonic development in Cynopterus sphinx during delayed development. The result showed the presence of a rate-limiting enzyme, ornithine decarboxylase-1, in both ovary and utero-embryonic unit of C. sphinx suggests a synthesis of putrescine in these sites. The corpus luteum showed increased, whereas utero-embryonic unit showed decreased production of putrescine during delayed development as compared with the normal development. The bat treated in vivo with putrescine during delayed development showed increase in progesterone and estradiol synthesis, correlated with increased expression of luteinizing hormone receptor, steroidogenic acute receptor protein, and 3β-hydroxysteroid dehydrogenase through extracellular signal-regulated kinase (ERK1/2)-mediated pathway in the ovary; but showed increase in the weight and expression of progesterone receptor (PR), B-cell lymphoma 2, proliferating cell nucleus antigen, and vascular endothelial growth factor proteins in utero-embryonic unit. The in vitro treatment of putrescine showed stimulatory whereas treatment with an inhibitor of putrescine, 2-difluoromethylornithine caused an inhibitory effect on ovarian progesterone synthesis and cell proliferation, and cell survival in the utero-embryonic unit. In conclusion, the putrescine showed two separate roles during embryonic diapause, high concentration of putrescine in the ovary may support corpus luteum and basal synthesis of progesterone, whereas a low level of putrescine causes retarded embryonic development by inhibiting cell proliferation in the utero-embryonic unit. The bat treated with putrescine either directly promotes cell proliferation, cell survival, and angiogenic activities or acts indirectly increasing PR on utero-embryonic unit thereby activating development in delayed embryo in C. sphinx.

Keywords:  cell proliferation; corpus luteum; delayed development; putrescine; steroidogenesis

DOI:  https://doi.org/10.1002/mrd.23281
J Cell Mol Med. 2019 Sep 29.

Caspase recruitment domain family member 10 regulates carbamoyl phosphate synthase 1 and promotes cancer growth in bladder cancer cells.

Xi Liu, Xiaotong Zhang, Jianbin Bi, Zhenhua Li, Zhe Zhang, Chuize Kong.

  Bladder cancer, which can be divided into non-muscle-invasive and muscle-invasive bladder cancer, is the most common urinary cancer in the United States. Caspase recruitment domain family member 10 (CARD10), also named CARD-containing MAGUK protein 3 (CARMA3), is a member of the CARMA family and may activate the nuclear factor kappa B (NF-κB) pathway. We utilized RNA sequencing and metabolic mass spectrometry to identify the molecular and metabolic feature of CARD10. The signalling pathway of CARD10 was verified by Western blotting analysis and functional assays. RNA sequencing and metabolic mass spectrometry of CARD10 knockdown identified the metabolic enzyme carbamoyl phosphate synthase 1 (CPS1) in the urea cycle as the downstream gene regulated by CARD10. We confirmed that CARD10 affected cell proliferation and nucleotide metabolism through regulating CPS1. We indicated that CARD10 promote bladder cancer growth via CPS1 and maybe a potential therapeutic target in bladder cancer.

Keywords:  CARD10; CPS1; NF-κB pathway; urinary bladder cancer

DOI:  https://doi.org/10.1111/jcmm.14683
Neurogastroenterol Motil. 2019 Sep 29. e13728

Functional involvement of acid-sensing ion channel 3 in the swallowing reflex in rats.

Mohammad Zakir Hossain, Hiroshi Ando, Shumpei Unno, Tetsuji Nakamoto, Junichi Kitagawa.

   BACKGROUND: Difficulty swallowing represents a major health problem. Swallowing function is improved by incorporating weak acids in suspensions/food boluses, implicating acid-sensing ion channels (ASICs) in the swallowing reflex. However, the functional involvement of ASICs in the swallowing reflex has not been fully elucidated.
METHODS: We localized ASIC3s in swallowing-related regions innervated by the superior laryngeal nerves (SLNs) and those in the nodose-petrosal-jugular ganglionic complex (NPJc) and examined their functional involvement in evoking the swallowing reflex in rats.
KEY RESULTS: We localized ASIC3s on epithelial cells and nerve fibers in swallowing-related regions innervated by the SLNs. In the NPJc, around half of the SLN-afferent neurons expressed ASIC3. Two-thirds of ASIC3s were localized on unmyelinated neurons in the nodose and petrosal ganglia. In the jugular ganglia, they were equally distributed on unmyelinated and myelinated neurons. Topical application of a synthetic non-proton ASIC3 activator, 2-guanidine-4-methylquinazoline (GMQ), and its natural endogenous ligand agmatine (a metabolite of the amino acid arginine) in swallowing-related regions evoked a considerable number of swallowing reflexes. Increasing the concentration of GMQ and agmatine up to a certain concentration increased the number of evoked reflexes and shortened the interval between the evoked reflexes. Agmatine was less potent than GMQ in its ability to evoke swallowing reflexes. Prior topical application of an ASIC3 antagonist significantly attenuated the number of GMQ- and agmatine-evoked swallowing reflexes.
CONCLUSIONS & INFERENCES: Acid-sensing ion channel 3s localized on nerves and epithelial cells in swallowing-related regions are functional in evoking the swallowing reflex and activation of these channels via a pharmacological agonist appears to improve swallowing behavior.

Keywords:  ASIC3; GMQ; agmatine; swallowing reflex

DOI:  https://doi.org/10.1111/nmo.13728