Foundation Publications

2021

  • S. Coni, F. A. Falconio, M. Marzullo, M. Munafò, B. Zuliani, F. Mosti, A. Fatica, Z. Ianniello, R. Bordone, A. Macone, E. Agostinelli, A. Perna, T. Matkovic, S. Sigrist, G. Silvestri, G. Canettieri, and L. Ciapponi, “Translational control of polyamine metabolism by CNBP is required for drosophila locomotor function,” eLife, vol. 10, 2021. doi:10.7554/elife.69269
    [BibTeX] [Abstract] [Download PDF]

    Microsatellite expansions of CCTG repeats in the cellular nucleic acid-binding protein (CNBP) gene leads to accumulation of toxic RNA and have been associated with myotonic dystrophy type 2 (DM2). However, it is still unclear whether the dystrophic phenotype is also linked to CNBP decrease, a conserved CCHC-type zinc finger RNA-binding protein that regulates translation and is required for mammalian development. Here, we show that depletion of Drosophila CNBP in muscles causes ageing-dependent locomotor defects that are correlated with impaired polyamine metabolism. We demonstrate that the levels of ornithine decarboxylase (ODC) and polyamines are significantly reduced upon dCNBP depletion. Of note, we show a reduction of the CNBP-polyamine axis in muscles from DM2 patients. Mechanistically, we provide evidence that dCNBP controls polyamine metabolism through binding dOdc mRNA and regulating its translation. Remarkably, the locomotor defect of dCNBP-deficient flies is rescued by either polyamine supplementation or dOdc1 overexpression. We suggest that this dCNBP function is evolutionarily conserved in vertebrates with relevant implications for CNBP-related pathophysiological conditions.

    @article{Coni2021,
    doi = {10.7554/elife.69269},
    url = {https://doi.org/10.7554/elife.69269},
    year = {2021},
    month = sep,
    publisher = {{eLife} Sciences Publications, Ltd},
    volume = {10},
    author = {Sonia Coni and Federica A Falconio and Marta Marzullo and Marzia Munaf{\`{o}} and Benedetta Zuliani and Federica Mosti and Alessandro Fatica and Zaira Ianniello and Rosa Bordone and Alberto Macone and Enzo Agostinelli and Alessia Perna and Tanja Matkovic and Stephan Sigrist and Gabriella Silvestri and Gianluca Canettieri and Laura Ciapponi},
    title = {Translational control of polyamine metabolism by {CNBP} is required for Drosophila locomotor function},
    journal = {{eLife}},
    abstract="Microsatellite expansions of CCTG repeats in the cellular nucleic acid-binding protein (CNBP) gene leads to accumulation of toxic RNA and have been associated with myotonic dystrophy type 2 (DM2). However, it is still unclear whether the dystrophic phenotype is also linked to CNBP decrease, a conserved CCHC-type zinc finger RNA-binding protein that regulates translation and is required for mammalian development. Here, we show that depletion of Drosophila CNBP in muscles causes ageing-dependent locomotor defects that are correlated with impaired polyamine metabolism. We demonstrate that the levels of ornithine decarboxylase (ODC) and polyamines are significantly reduced upon dCNBP depletion. Of note, we show a reduction of the CNBP-polyamine axis in muscles from DM2 patients. Mechanistically, we provide evidence that dCNBP controls polyamine metabolism through binding dOdc mRNA and regulating its translation. Remarkably, the locomotor defect of dCNBP-deficient flies is rescued by either polyamine supplementation or dOdc1 overexpression. We suggest that this dCNBP function is evolutionarily conserved in vertebrates with relevant implications for CNBP-related pathophysiological conditions."
    }

  • Y. Kanamori, A. Finotti, L. D. Magno, G. Canettieri, T. Tahara, F. Timeus, A. Greco, P. Tirassa, J. Gasparello, P. Fino, C. D. Liegro, P. Proia, G. Schiera, I. D. Liegro, R. Gambari, and E. Agostinelli, “Enzymatic spermine metabolites induce apoptosis associated with increase of p53, caspase-3 and miR-34a in both neuroblastoma cells, SJNKP and the n-myc-amplified form IMR5,” , vol. 10, iss. 8, p. 1950, 2021. doi:10.3390/cells10081950
    [BibTeX] [Abstract] [Download PDF]

    Neuroblastoma (NB) is a common malignant solid tumor in children and accounts for 15% of childhood cancer mortality. Amplification of the N-Myc oncogene is a well-established poor prognostic marker in NB patients and strongly correlates with higher tumor aggression and resistance to treatment. New therapies for patients with N-Myc-amplified NB need to be developed. After treating NB cells with BSAO/SPM, the detection of apoptosis was determined after annexin V-FITC labeling and DNA staining with propidium iodide. The mitochondrial membrane potential activity was checked, labeling cells with the probe JC-1 dye. We analyzed, by real-time RT-PCR, the transcript of genes involved in the apoptotic process, to determine possible down- or upregulation of mRNAs after the treatment on SJNKP and the N-Myc-amplified IMR5 cell lines with BSAO/SPM. The experiments were carried out considering the proapoptotic genes Tp53 and caspase-3. After treatment with BSAO/SPM, both cell lines displayed increased mRNA levels for all these proapoptotic genes. Western blotting analysis with PARP and caspase-3 antibody support that BSAO/SPM treatment induces high levels of apoptosis in cells. The major conclusion is that BSAO/SPM treatment leads to antiproliferative and cytotoxic activity of both NB cell lines, associated with activation of apoptosis.

    @Article{yuta,
    doi = {10.3390/cells10081950},
    url = {https://doi.org/10.3390/cells10081950},
    year = {2021},
    month = jul,
    publisher = {{MDPI} {AG}},
    volume = {10},
    number = {8},
    pages = {1950},
    author = {Yuta Kanamori and Alessia Finotti and Laura Di Magno and Gianluca Canettieri and Tomoaki Tahara and Fabio Timeus and Antonio Greco and Paola Tirassa and Jessica Gasparello and Pasquale Fino and Carlo Di Liegro and Patrizia Proia and Gabriella Schiera and Italia Di Liegro and Roberto Gambari and Enzo Agostinelli},
    title = {Enzymatic Spermine Metabolites Induce Apoptosis Associated with Increase of p53, caspase-3 and {miR}-34a in Both Neuroblastoma Cells, {SJNKP} and the N-Myc-Amplified Form {IMR}5},
    abstract="Neuroblastoma (NB) is a common malignant solid tumor in children and accounts for 15% of childhood cancer mortality. Amplification of the N-Myc oncogene is a well-established poor prognostic marker in NB patients and strongly correlates with higher tumor aggression and resistance to treatment. New therapies for patients with N-Myc-amplified NB need to be developed. After treating NB cells with BSAO/SPM, the detection of apoptosis was determined after annexin V-FITC labeling and DNA staining with propidium iodide. The mitochondrial membrane potential activity was checked, labeling cells with the probe JC-1 dye. We analyzed, by real-time RT-PCR, the transcript of genes involved in the apoptotic process, to determine possible down- or upregulation of mRNAs after the treatment on SJNKP and the N-Myc-amplified IMR5 cell lines with BSAO/SPM. The experiments were carried out considering the proapoptotic genes Tp53 and caspase-3. After treatment with BSAO/SPM, both cell lines displayed increased mRNA levels for all these proapoptotic genes. Western blotting analysis with PARP and caspase-3 antibody support that BSAO/SPM treatment induces high levels of apoptosis in cells. The major conclusion is that BSAO/SPM treatment leads to antiproliferative and cytotoxic activity of both NB cell lines, associated with activation of apoptosis."
    }

  • I. Pottosin, M. Olivas-Aguirre, O. Dobrovinskaya, I. Zepeda-Jazo, and S. Shabala, “Modulation of ion transport across plant membranes by polyamines: understanding specific modes of action under stress,” , vol. 11, 2021. doi:10.3389/fpls.2020.616077
    [BibTeX] [Abstract] [Download PDF]

    This work critically discusses the direct and indirect effects of natural polyamines and their catabolites such as reactive oxygen species and γ-aminobutyric acid on the activity of key plant ion-transporting proteins such as plasma membrane H+ and Ca2+ ATPases and K+-selective and cation channels in the plasma membrane and tonoplast, in the context of their involvement in stress responses. Docking analysis predicts a distinct binding for putrescine and longer polyamines within the pore of the vacuolar TPC1/SV channel, one of the key determinants of the cell ionic homeostasis and signaling under stress conditions, and an additional site for spermine, which overlaps with the cytosolic regulatory Ca2+-binding site. Several unresolved problems are summarized, including the correct estimates of the subcellular levels of polyamines and their catabolites, their unexplored effects on nucleotide-gated and glutamate receptor channels of cell membranes and Ca2+-permeable and K+-selective channels in the membranes of plant mitochondria and chloroplasts, and pleiotropic mechanisms of polyamines’ action on H+ and Ca2+ pumps.

    @Article{igor,
    doi = {10.3389/fpls.2020.616077},
    url = {https://doi.org/10.3389/fpls.2020.616077},
    year = {2021},
    month = jan,
    publisher = {Frontiers Media {SA}},
    volume = {11},
    author = {Igor Pottosin and Miguel Olivas-Aguirre and Oxana Dobrovinskaya and Isaac Zepeda-Jazo and Sergey Shabala},
    title = {Modulation of Ion Transport Across Plant Membranes by Polyamines: Understanding Specific Modes of Action Under Stress},
    abstract="This work critically discusses the direct and indirect effects of natural polyamines and their catabolites such as reactive oxygen species and γ-aminobutyric acid on the activity of key plant ion-transporting proteins such as plasma membrane H+ and Ca2+ ATPases and K+-selective and cation channels in the plasma membrane and tonoplast, in the context of their involvement in stress responses. Docking analysis predicts a distinct binding for putrescine and longer polyamines within the pore of the vacuolar TPC1/SV channel, one of the key determinants of the cell ionic homeostasis and signaling under stress conditions, and an additional site for spermine, which overlaps with the cytosolic regulatory Ca2+-binding site. Several unresolved problems are summarized, including the correct estimates of the subcellular levels of polyamines and their catabolites, their unexplored effects on nucleotide-gated and glutamate receptor channels of cell membranes and Ca2+-permeable and K+-selective channels in the membranes of plant mitochondria and chloroplasts, and pleiotropic mechanisms of polyamines’ action on H+ and Ca2+ pumps."
    }

2020

  • E. Agostinelli, “Biochemical and pathophysiological properties of polyamines,” , vol. 52, iss. 2, p. 111–117, 2020. doi:10.1007/s00726-020-02821-8
    [BibTeX] [Abstract] [Download PDF]

    The history of polyamines dates back to the fifteenth century when spermine was discovered by Antonie van Leeuwenhoek [born in Delft, Holland (1632–1723)], but it took several decades before scientists got interested in understanding and unraveling the role(s) of spermine and other polyamines in the biology of living cells. Mammalian cells contain significant amounts of polyamines and these molecules, which are polycations, play specific roles in various tissues. Although the physiological functions of these polycations have yet to be elucidated completely at the molecular level, many studies have provided a better understanding of the roles polyamines play in cell growth, proliferation, and pathophysiological processes. At the 5th International Conference on Polyamines: Biochemical, Physiological and Clinical Perspectives held in Taiwan, in 2018, special attention has been given to the role of polyamines in carcinogenesis and in developing new approaches for cancer therapy and other diseases. The issue is a tribute and dedicated by internationally recognized experts to the memory of Professor Seymour S. Cohen, a prominent scientist in polyamine research. The manuscripts included in this special issue range from biochemistry to pharmacology, chemistry, genetics, molecular biology and clinical science on the current state of knowledge regarding the physiological, biochemical, and therapeutic actions of polyamines, and should be of use to the old and the new generation of researchers in the polyamine field.

    @Article{enzo,
    doi = {10.1007/s00726-020-02821-8},
    url = {https://doi.org/10.1007/s00726-020-02821-8},
    year = {2020},
    month = feb,
    publisher = {Springer Science and Business Media {LLC}},
    volume = {52},
    number = {2},
    pages = {111--117},
    author = {Enzo Agostinelli},
    title = {Biochemical and pathophysiological properties of polyamines},
    abstract="The history of polyamines dates back to the fifteenth century when spermine was discovered by Antonie van Leeuwenhoek [born in Delft, Holland (1632–1723)], but it took several decades before scientists got interested in understanding and unraveling the role(s) of spermine and other polyamines in the biology of living cells. Mammalian cells contain significant amounts of polyamines and these molecules, which are polycations, play specific roles in various tissues. Although the physiological functions of these polycations have yet to be elucidated completely at the molecular level, many studies have provided a better understanding of the roles polyamines play in cell growth, proliferation, and pathophysiological processes. At the 5th International Conference on Polyamines: Biochemical, Physiological and Clinical Perspectives held in Taiwan, in 2018, special attention has been given to the role of polyamines in carcinogenesis and in developing new approaches for cancer therapy and other diseases.
    The issue is a tribute and dedicated by internationally recognized experts to the memory of Professor Seymour S. Cohen, a prominent scientist in polyamine research. The manuscripts included in this special issue range from biochemistry to pharmacology, chemistry, genetics, molecular biology and clinical science on the current state of knowledge regarding the physiological, biochemical, and therapeutic actions of polyamines, and should be of use to the old and the new generation of researchers in the polyamine field."
    }

  • S. Coni, S. M. Serrao, Z. N. Yurtsever, L. D. Magno, R. Bordone, C. Bertani, V. Licursi, Z. Ianniello, P. Infante, M. Moretti, M. Petroni, F. Guerrieri, A. Fatica, A. Macone, E. D. Smaele, L. D. Marcotullio, G. Giannini, M. Maroder, E. Agostinelli, and G. Canettieri, “Blockade of EIF5a hypusination limits colorectal cancer growth by inhibiting MYC elongation,” , vol. 11, iss. 12, 2020. doi:10.1038/s41419-020-03174-6
    [BibTeX] [Abstract] [Download PDF]

    Eukaryotic Translation Initiation Factor 5A (EIF5A) is a translation factor regulated by hypusination, a unique posttranslational modification catalyzed by deoxyhypusine synthetase (DHPS) and deoxyhypusine hydroxylase (DOHH) starting from the polyamine spermidine. Emerging data are showing that hypusinated EIF5A regulates key cellular processes such as autophagy, senescence, polyamine homeostasis, energy metabolism, and plays a role in cancer. However, the effects of EIF5A inhibition in preclinical cancer models, the mechanism of action, and specific translational targets are still poorly understood. We show here that hypusinated EIF5A promotes growth of colorectal cancer (CRC) cells by directly regulating MYC biosynthesis at specific pausing motifs. Inhibition of EIF5A hypusination with the DHPS inhibitor GC7 or through lentiviral-mediated knockdown of DHPS or EIF5A reduces the growth of various CRC cells. Multiplex gene expression analysis reveals that inhibition of hypusination impairs the expression of transcripts regulated by MYC, suggesting the involvement of this oncogene in the observed effect. Indeed, we demonstrate that EIF5A regulates MYC elongation without affecting its mRNA content or protein stability, by alleviating ribosome stalling at five distinct pausing motifs in MYC CDS. Of note, we show that blockade of the hypusination axis elicits a remarkable growth inhibitory effect in preclinical models of CRC and significantly reduces the size of polyps in APCMin/+ mice, a model of human familial adenomatous polyposis (FAP). Together, these data illustrate an unprecedented mechanism, whereby the tumor-promoting properties of hypusinated EIF5A are linked to its ability to regulate MYC elongation and provide a rationale for the use of DHPS/EIF5A inhibitors in CRC therapy.

    @Article{sonia,
    doi = {10.1038/s41419-020-03174-6},
    url = {https://doi.org/10.1038/s41419-020-03174-6},
    year = {2020},
    month = dec,
    publisher = {Springer Science and Business Media {LLC}},
    volume = {11},
    number = {12},
    author = {Sonia Coni and Silvia Maria Serrao and Zuleyha Nihan Yurtsever and Laura Di Magno and Rosa Bordone and Camilla Bertani and Valerio Licursi and Zaira Ianniello and Paola Infante and Marta Moretti and Marialaura Petroni and Francesca Guerrieri and Alessandro Fatica and Alberto Macone and Enrico De Smaele and Lucia Di Marcotullio and Giuseppe Giannini and Marella Maroder and Enzo Agostinelli and Gianluca Canettieri},
    title = {Blockade of {EIF}5A hypusination limits colorectal cancer growth by inhibiting {MYC} elongation},
    abstract="Eukaryotic Translation Initiation Factor 5A (EIF5A) is a translation factor regulated by hypusination, a unique posttranslational modification catalyzed by deoxyhypusine synthetase (DHPS) and deoxyhypusine hydroxylase (DOHH) starting from the polyamine spermidine. Emerging data are showing that hypusinated EIF5A regulates key cellular processes such as autophagy, senescence, polyamine homeostasis, energy metabolism, and plays a role in cancer. However, the effects of EIF5A inhibition in preclinical cancer models, the mechanism of action, and specific translational targets are still poorly understood. We show here that hypusinated EIF5A promotes growth of colorectal cancer (CRC) cells by directly regulating MYC biosynthesis at specific pausing motifs. Inhibition of EIF5A hypusination with the DHPS inhibitor GC7 or through lentiviral-mediated knockdown of DHPS or EIF5A reduces the growth of various CRC cells. Multiplex gene expression analysis reveals that inhibition of hypusination impairs the expression of transcripts regulated by MYC, suggesting the involvement of this oncogene in the observed effect. Indeed, we demonstrate that EIF5A regulates MYC elongation without affecting its mRNA content or protein stability, by alleviating ribosome stalling at five distinct pausing motifs in MYC CDS. Of note, we show that blockade of the hypusination axis elicits a remarkable growth inhibitory effect in preclinical models of CRC and significantly reduces the size of polyps in APCMin/+ mice, a model of human familial adenomatous polyposis (FAP). Together, these data illustrate an unprecedented mechanism, whereby the tumor-promoting properties of hypusinated EIF5A are linked to its ability to regulate MYC elongation and provide a rationale for the use of DHPS/EIF5A inhibitors in CRC therapy."
    }

2019

  • F. Gasperoni, P. Turini, and E. Agostinelli, “A novel comprehensive paradigm for the etiopathogenesis of multiple sclerosis: therapeutic approaches and future perspectives on its treatment,” Amino acids, vol. 51, iss. 5, p. 745–759, 2019. doi:10.1007/s00726-019-02718-1
    [BibTeX] [Abstract] [Download PDF]

    It is well recognized that variation in the geographical distribution of prevalence of multiple sclerosis (MS) exists: increasing the latitude its prevalence increases as well, but the underlying causes of such dissimilarity still remained elusive as of today. Currently, the most accredited hypothesis is that the closer to the equator the more pronounced is the amount of sunlight which, in turn, increases the production of vitamin D. Cholecalciferol is indeed deficient in MS patients, but this factor does not explain by itself the etiopathogenesis of the disease. In the present study, to search for a pattern and provide a model of the disease’s etiology consistent with this regional factor, as well with its changing ethnic, sex-ratio, lifestyle variations and the other unexplained aspects of MS, an extensive analysis of peer-reviewed literature and data was conducted. The arisen hypothesis was that, increasing the latitude, the factor that varies and can have the stronger effect on the human organism, is the continuous and ever-increasing diversity of the natural light–dark cycle. The consequent effort of the suprachiasmatic nucleus to entrain the organism’s circadian rhythm affects the hypothalamic–pituitary–adrenal axis resulting in desynchronizing the central and peripheral circadian clocks and pathologizing the immunitary system. To verify such hypothesis, a theoretical framework of the etiopathogenesis, coherent with the gathered literature, was conceived and a demonstration to corroborate it was eventually devised and performed. The results underscored that people living in countries subjected to a further circadian disruptive factor, as daylight saving time, have a 6.35 times higher prevalence of MS than States placed on their same latitude that do not observe it, thus strongly supporting the hypothesis. As further reinforcement of the conclusions, it is worth mentioning that the levels of polyamines rise abruptly in autoimmune diseases. Moreover, among their numerous roles, these polycations participate to the regulation of the circadian clock so their sudden variation might disrupt it. Following these interesting findings, new perspectives in therapies are, therefore, proposed.

    @Article{Gasperoni2019,
    author="Gasperoni, Francesco
    and Turini, Paola
    and Agostinelli, Enzo",
    title="A novel comprehensive paradigm for the etiopathogenesis of multiple sclerosis: therapeutic approaches and future perspectives on its treatment",
    journal="Amino Acids",
    year="2019",
    month="May",
    day="01",
    volume="51",
    number="5",
    pages="745--759",
    abstract="It is well recognized that variation in the geographical distribution of prevalence of multiple sclerosis (MS) exists: increasing the latitude its prevalence increases as well, but the underlying causes of such dissimilarity still remained elusive as of today. Currently, the most accredited hypothesis is that the closer to the equator the more pronounced is the amount of sunlight which, in turn, increases the production of vitamin D. Cholecalciferol is indeed deficient in MS patients, but this factor does not explain by itself the etiopathogenesis of the disease. In the present study, to search for a pattern and provide a model of the disease's etiology consistent with this regional factor, as well with its changing ethnic, sex-ratio, lifestyle variations and the other unexplained aspects of MS, an extensive analysis of peer-reviewed literature and data was conducted. The arisen hypothesis was that, increasing the latitude, the factor that varies and can have the stronger effect on the human organism, is the continuous and ever-increasing diversity of the natural light--dark cycle. The consequent effort of the suprachiasmatic nucleus to entrain the organism's circadian rhythm affects the hypothalamic--pituitary--adrenal axis resulting in desynchronizing the central and peripheral circadian clocks and pathologizing the immunitary system. To verify such hypothesis, a theoretical framework of the etiopathogenesis, coherent with the gathered literature, was conceived and a demonstration to corroborate it was eventually devised and performed. The results underscored that people living in countries subjected to a further circadian disruptive factor, as daylight saving time, have a 6.35 times higher prevalence of MS than States placed on their same latitude that do not observe it, thus strongly supporting the hypothesis. As further reinforcement of the conclusions, it is worth mentioning that the levels of polyamines rise abruptly in autoimmune diseases. Moreover, among their numerous roles, these polycations participate to the regulation of the circadian clock so their sudden variation might disrupt it. Following these interesting findings, new perspectives in therapies are, therefore, proposed.",
    issn="1438-2199",
    doi="10.1007/s00726-019-02718-1",
    url="https://doi.org/10.1007/s00726-019-02718-1"
    }

  • S. Coni, L. Di Magno, S. M. Serrao, Y. Kanamori, E. Agostinelli, and G. Canettieri, “Polyamine metabolism as a therapeutic target in hedgehog-driven basal cell carcinoma and medulloblastoma,” Cells, vol. 8, iss. 2, 2019. doi:10.3390/cells8020150
    [BibTeX] [Abstract] [Download PDF]

    Hedgehog (Hh) signaling is a critical developmental regulator and its aberrant activation, due to somatic or germline mutations of genes encoding pathway components, causes Basal Cell Carcinoma (BCC) and medulloblastoma (MB). A growing effort has been devoted at the identification of druggable vulnerabilities of the Hedgehog signaling, leading to the identification of various compounds with variable efficacy and/or safety. Emerging evidence shows that an aberrant polyamine metabolism is a hallmark of Hh-dependent tumors and that its pharmacological inhibition elicits relevant therapeutic effects in clinical or preclinical models of BCC and MB. We discuss here the current knowledge of polyamine metabolism, its role in cancer and the available targeting strategies. We review the literature about the connection between polyamines and the Hedgehog signaling, and the potential therapeutic benefit of targeting polyamine metabolism in two malignancies where Hh pathways play a well-established role: BCC and MB.

    @Article{cells8020150,
    AUTHOR = {Coni, Sonia and Di Magno, Laura and Serrao, Silvia Maria and Kanamori, Yuta and Agostinelli, Enzo and Canettieri, Gianluca},
    TITLE = {Polyamine Metabolism as a Therapeutic Target in Hedgehog-Driven Basal Cell Carcinoma and Medulloblastoma},
    JOURNAL = {Cells},
    VOLUME = {8},
    YEAR = {2019},
    NUMBER = {2},
    ARTICLE-NUMBER = {150},
    URL = {https://www.mdpi.com/2073-4409/8/2/150},
    PubMedID = {30754726},
    ISSN = {2073-4409},
    ABSTRACT = {Hedgehog (Hh) signaling is a critical developmental regulator and its aberrant activation, due to somatic or germline mutations of genes encoding pathway components, causes Basal Cell Carcinoma (BCC) and medulloblastoma (MB). A growing effort has been devoted at the identification of druggable vulnerabilities of the Hedgehog signaling, leading to the identification of various compounds with variable efficacy and/or safety. Emerging evidence shows that an aberrant polyamine metabolism is a hallmark of Hh-dependent tumors and that its pharmacological inhibition elicits relevant therapeutic effects in clinical or preclinical models of BCC and MB. We discuss here the current knowledge of polyamine metabolism, its role in cancer and the available targeting strategies. We review the literature about the connection between polyamines and the Hedgehog signaling, and the potential therapeutic benefit of targeting polyamine metabolism in two malignancies where Hh pathways play a well-established role: BCC and MB.},
    DOI = {10.3390/cells8020150}
    }

  • M. L. Di Paolo, M. Cervelli, P. Mariottini, A. Leonetti, F. Polticelli, M. Rosini, A. Milelli, F. Basagni, R. Venerando, E. Agostinelli, and A. Minarini, “Exploring the activity of polyamine analogues on polyamine and spermine oxidase: methoctramine, a potent and selective inhibitor of polyamine oxidase,” Journal of enzyme inhibition and medicinal chemistry, pp. 740-752, 2019. doi:10.1080/14756366.2019.1584620
    [BibTeX] [Abstract] [Download PDF]

    Fourteen polyamine analogues, asymmetric or symmetric substituted spermine (1–9) or methoctramine (10–14) analogues, were evaluated as potential inhibitors or substrates of two enzymes of the polyamine catabolic pathway, spermine oxidase (SMOX) and acetylpolyamine oxidase (PAOX). Compound 2 turned out to be the best substrate for PAOX, having the highest affinity and catalytic efficiency with respect to its physiological substrates. Methoctramine (10), a well-known muscarinic M2 receptor antagonist, emerged as the most potent competitive PAOX inhibitor known so far (Ki = 10 nM), endowed with very good selectivity compared with SMOX (Ki=1.2 μM vs SMOX). The efficacy of methoctramine in inhibiting PAOX activity was confirmed in the HT22 cell line. Methoctramine is a very promising tool in the design of drugs targeting the polyamine catabolism pathway, both to understand the physio-pathological role of PAOX vs SMOX and for pharmacological applications, being the polyamine pathway involved in various pathologies.

    @Article{dipaolo,
    author = {Di Paolo, Maria Luisa and Cervelli, Manuela and Mariottini, Paolo and Leonetti, Alessia and Polticelli, Fabio and Rosini, Michela and Milelli, Andrea and Basagni, Filippo and Venerando, Rina and Agostinelli, Enzo and Minarini, Anna},
    year = {2019},
    month = {03},
    pages = {740-752},
    title = {Exploring the activity of polyamine analogues on polyamine and spermine oxidase: methoctramine, a potent and selective inhibitor of polyamine oxidase},
    url="https://doi.org/10.1080/14756366.2019.1584620"
    volume = {34},
    journal = {Journal of Enzyme Inhibition and Medicinal Chemistry},
    doi = {10.1080/14756366.2019.1584620},
    abstract="Fourteen polyamine analogues, asymmetric or symmetric substituted spermine (1–9) or methoctramine (10–14) analogues, were evaluated as potential inhibitors or substrates of two enzymes of the polyamine catabolic pathway, spermine oxidase (SMOX) and acetylpolyamine oxidase (PAOX). Compound 2 turned out to be the best substrate for PAOX, having the highest affinity and catalytic efficiency with respect to its physiological substrates. Methoctramine (10), a well-known muscarinic M2 receptor antagonist, emerged as the most potent competitive PAOX inhibitor known so far (Ki = 10 nM), endowed with very good selectivity compared with SMOX (Ki=1.2 μM vs SMOX). The efficacy of methoctramine in inhibiting PAOX activity was confirmed in the HT22 cell line. Methoctramine is a very promising tool in the design of drugs targeting the polyamine catabolism pathway, both to understand the physio-pathological role of PAOX vs SMOX and for pharmacological applications, being the polyamine pathway involved in various pathologies."
    }

  • S. Ohkubo, R. Mancinelli, S. Miglietta, A. Cona, R. Angelini, G. Canettieri, D. A Spandidos, E. Gaudio, and E. Agostinelli, “Maize polyamine oxidase in the presence of spermine/spermidine induces the apoptosis of lovo human colon adenocarcinoma cells,” International journal of oncology, vol. 54, 2019. doi:10.3892/ijo.2019.4780
    [BibTeX] [Abstract] [Download PDF]

    Amine oxidases, which contribute to the regulation of polyamine levels, catalyze the oxidative deamination of polyamines to generate H2O2 and aldehyde(s). In this study, and at least to the best of our knowledge, maize polyamine oxidase (ZmPAO) was used for the first time with the aim of identifying a novel strategy for cancer therapy. The cytotoxicity and the mechanisms of cell death induced by the enzymatic oxidation products of polyamine generated by ZmPAO were investigated. Exogenous spermine and ZmPAO treatment decreased cell viability in a spermine dose‑ and time‑dependent manner, particularly, the viability of the multidrug‑resistant (MDR) colon adenocarcinoma cells, LoVo DX, when compared with drug‑sensitive ones (LoVo WT). Further analyses revealed that H2O2 derived from spermine was mainly responsible for the cytotoxicity. Flow cytometric analysis revealed that treatment with ZmPAO and spermine increased the apoptotic population of LoVo WT and LoVo DX cells. In addition, we found that treatment with ZmPAO and spermine markedly reduced mitochondrial membrane potential in the LoVo DX cells, in agreement with the results of cell viability and apoptosis assays. Transmission electron microscopic observations supported the involvement of mitochondrial depolarization in the apoptotic process. Therefore, the dysregulation of polyamine metabolism in tumor cells may be a potential therapeutic target. In addition, the development of MDR tumor cells is recognized as a major obstacle in cancer therapy. Therefore, the design of a novel therapeutic strategy based on the use of this combination may be taken into account, making this approach attractive mainly in treating MDR cancer patients.

    @Article{ohkubo,
    author = {Ohkubo, Shinji and Mancinelli, Romina and Miglietta, Selenia and Cona, Alessandra and Angelini, Riccardo and Canettieri, Gianluca and A Spandidos, Demetrios and Gaudio, Eugenio and Agostinelli, Enzo},
    year = {2019},
    month = {04},
    pages = {},
    title = {Maize polyamine oxidase in the presence of spermine/spermidine induces the apoptosis of LoVo human colon adenocarcinoma cells},
    volume = {54},
    journal = {International Journal of Oncology},
    doi = {10.3892/ijo.2019.4780},
    url="https://doi.org/10.3892/ijo.2019.4780",
    abstract="Amine oxidases, which contribute to the regulation of polyamine levels, catalyze the oxidative deamination of polyamines to generate H2O2 and aldehyde(s). In this study, and at least to the best of our knowledge, maize polyamine oxidase (ZmPAO) was used for the first time with the aim of identifying a novel strategy for cancer therapy. The cytotoxicity and the mechanisms of cell death induced by the enzymatic oxidation products of polyamine generated by ZmPAO were investigated. Exogenous spermine and ZmPAO treatment decreased cell viability in a spermine dose‑ and time‑dependent manner, particularly, the viability of the multidrug‑resistant (MDR) colon adenocarcinoma cells, LoVo DX, when compared with drug‑sensitive ones (LoVo WT). Further analyses revealed that H2O2 derived from spermine was mainly responsible for the cytotoxicity. Flow cytometric analysis revealed that treatment with ZmPAO and spermine increased the apoptotic population of LoVo WT and LoVo DX cells. In addition, we found that treatment with ZmPAO and spermine markedly reduced mitochondrial membrane potential in the LoVo DX cells, in agreement with the results of cell viability and apoptosis assays. Transmission electron microscopic observations supported the involvement of mitochondrial depolarization in the apoptotic process. Therefore, the dysregulation of polyamine metabolism in tumor cells may be a potential therapeutic target. In addition, the development of MDR tumor cells is recognized as a major obstacle in cancer therapy. Therefore, the design of a novel therapeutic strategy based on the use of this combination may be taken into account, making this approach attractive mainly in treating MDR cancer patients.
    "
    }

  • M. Magro, D. Baratella, G. Miotto, J. Frömmel, M. Šebela, M. Kopečná, E. Agostinelli, and F. Vianello, “Enzyme self-assembly on naked iron oxide nanoparticles for aminoaldehyde biosensing,” Amino acids, vol. 51, iss. 4, p. 679–690, 2019. doi:10.1007/s00726-019-02704-7
    [BibTeX] [Abstract] [Download PDF]

    The preservation of enzymatic activity is a fundamental requirement for exploiting hybrid nano-bio-conjugates, and the control over protein–nanopArticle interactions, leading to stable and catalytically active hybrids, represents the key for designing new biosensing platforms. In this scenario, surface active maghemite nanopArticles (SAMNs) represent a new class of naked magnetic nanopArticles, displaying peculiar electrocatalytic features and the ability to selectively bind proteins. Recombinant aminoaldehyde dehydrogenase from tomato (SlAMADH1) was used as a model protein, and successfully immobilized by self-assembly on the surface of naked SAMNs, where its enzymatic activity resulted preserved for more than 6 months. The hybrid nanomaterial (SAMN@SlAMADH1) was characterized by UV–Vis spectroscopy, mass spectrometry, and TEM microscopy, and applied for the development of a biosensor for the determination of aminoaldehydes in alcoholic beverages. Measurements were carried out in a low volume electrochemical flow cell comprising a SAMN modified carbon paste electrode for the coulometric determination of the NADH produced during the enzymatic catalysis. The present findings, besides representing the first example of an electrochemical biosensor for aminoaldehydes in an alcoholic matrix, open the door to the use of immobilized enzymes on naked metal oxides nanomaterials for biosensing.

    @Article{Magro2019,
    author="Magro, Massimiliano
    and Baratella, Davide
    and Miotto, Giovanni
    and Fr{\"o}mmel, Jan
    and {\v{S}}ebela, Marek
    and Kope{\v{c}}n{\'a}, Martina
    and Agostinelli, Enzo
    and Vianello, Fabio",
    title="Enzyme self-assembly on naked iron oxide nanopArticles for aminoaldehyde biosensing",
    journal="Amino Acids",
    year="2019",
    month="Apr",
    day="01",
    volume="51",
    number="4",
    pages="679--690",
    abstract="The preservation of enzymatic activity is a fundamental requirement for exploiting hybrid nano-bio-conjugates, and the control over protein--nanopArticle interactions, leading to stable and catalytically active hybrids, represents the key for designing new biosensing platforms. In this scenario, surface active maghemite nanopArticles (SAMNs) represent a new class of naked magnetic nanopArticles, displaying peculiar electrocatalytic features and the ability to selectively bind proteins. Recombinant aminoaldehyde dehydrogenase from tomato (SlAMADH1) was used as a model protein, and successfully immobilized by self-assembly on the surface of naked SAMNs, where its enzymatic activity resulted preserved for more than 6 months. The hybrid nanomaterial (SAMN@SlAMADH1) was characterized by UV--Vis spectroscopy, mass spectrometry, and TEM microscopy, and applied for the development of a biosensor for the determination of aminoaldehydes in alcoholic beverages. Measurements were carried out in a low volume electrochemical flow cell comprising a SAMN modified carbon paste electrode for the coulometric determination of the NADH produced during the enzymatic catalysis. The present findings, besides representing the first example of an electrochemical biosensor for aminoaldehydes in an alcoholic matrix, open the door to the use of immobilized enzymes on naked metal oxides nanomaterials for biosensing.",
    issn="1438-2199",
    doi="10.1007/s00726-019-02704-7",
    url="https://doi.org/10.1007/s00726-019-02704-7"
    }

  • P. Martinis, S. Grancara, Y. Kanamori, A. N. García-Argáez, E. Pacella, L. D. Via, A. Toninello, and E. Agostinelli, “Involvement of the biogenic active amine agmatine in mitochondrial membrane permeabilization and release of pro-apoptotic factors,” , vol. 52, iss. 2, p. 161–169, 2019. doi:10.1007/s00726-019-02791-6
    [BibTeX] [Abstract] [Download PDF]

    Agmatine (AGM) produces a dual effect on the mitochondrial permeability transition (MPT) mechanism in rat liver mitochondria: at low concentrations, it induces the phenomenon, at high ones, inhibits it. The prevention at high concentrations is evidenced by the significant inhibition of mitochondrial swelling induced by Ca2+ and phosphate; in this condition, AGM both prevents the release of Apoptosis Inducing Factor (AIF) and enhances the release of other pro-apoptotic factors, such as cytochrome c (cyt c) and Smac/DIABLO. As these factors are released without MPT induction, the involvement of mitochondrial outer membrane permeabilization (MOMP) could be hypothesized. Cyclosporin A (CsA), a powerful inhibitor of MPT, enhanced the AGM-mediated inhibition of swelling, and surprisingly, prevented the release of cyt c and Smac/DIABLO. In the presence of Ca2+, AGM also activated the Bcl-2 family protein Bax, a key factor in inducing MOMP, which is inactivated by CsA. Together with the voltage-dependent anion channel (VDAC), Bax forms channels in the outer membrane further supporting the involvement of MOMP in the release of pro-apoptotic factors. In view of the fact that VDAC was inactivated by ruthenium red, which in turn inhibited the release of cyt c, it can be hypothesized that, on the one hand, AGM inhibits MPT induction and, on the other, it selectively permeabilizes the outer membrane via MOMP induction.

    @Article{pamela,
    doi = {10.1007/s00726-019-02791-6},
    url = {https://doi.org/10.1007/s00726-019-02791-6},
    year = {2019},
    month = oct,
    publisher = {Springer Science and Business Media {LLC}},
    volume = {52},
    number = {2},
    pages = {161--169},
    author = {Pamela Martinis and Silvia Grancara and Yuta Kanamori and A{\'{\i}}da Nelly Garc{\'{\i}}a-Arg{\'{a}}ez and Elena Pacella and Lisa Dalla Via and Antonio Toninello and Enzo Agostinelli},
    title = {Involvement of the biogenic active amine agmatine in mitochondrial membrane permeabilization and release of pro-apoptotic factors},
    abstract="Agmatine (AGM) produces a dual effect on the mitochondrial permeability transition (MPT) mechanism in rat liver mitochondria: at low concentrations, it induces the phenomenon, at high ones, inhibits it. The prevention at high concentrations is evidenced by the significant inhibition of mitochondrial swelling induced by Ca2+ and phosphate; in this condition, AGM both prevents the release of Apoptosis Inducing Factor (AIF) and enhances the release of other pro-apoptotic factors, such as cytochrome c (cyt c) and Smac/DIABLO. As these factors are released without MPT induction, the involvement of mitochondrial outer membrane permeabilization (MOMP) could be hypothesized. Cyclosporin A (CsA), a powerful inhibitor of MPT, enhanced the AGM-mediated inhibition of swelling, and surprisingly, prevented the release of cyt c and Smac/DIABLO. In the presence of Ca2+, AGM also activated the Bcl-2 family protein Bax, a key factor in inducing MOMP, which is inactivated by CsA. Together with the voltage-dependent anion channel (VDAC), Bax forms channels in the outer membrane further supporting the involvement of MOMP in the release of pro-apoptotic factors. In view of the fact that VDAC was inactivated by ruthenium red, which in turn inhibited the release of cyt c, it can be hypothesized that, on the one hand, AGM inhibits MPT induction and, on the other, it selectively permeabilizes the outer membrane via MOMP induction."
    }