Effects in cancer cells of the recombinant l-Methionine Gamma-Lyase from Brevibacterium aurantiacum. Encapsulation in human erythrocytes for sustained l-Methionine Elimination

Autor: E. Goldschmidt, Raphaël Saffroy, Julien Dairou, David Kopečný, Pascal Bonnarme, Pierre Briozzo, Luigia Rossi, Bernadette Chadefaux-Vekemans, Mauro Magnani, Francesca Pierigè, Rosella Mollicone, Jocelyne Hamelin, Claude Boucheix, Yann Gaston-Mathé, Farhad Haghighi-Rad, David Machover, Christophe Desterke
Přispěvatelé: Hôpital Paul Brousse, Modèles de Cellules Souches Malignes et Thérapeutiques, Université Paris-Sud - Paris 11 (UP11)-Institut National de la Santé et de la Recherche Médicale (INSERM), Université Paris-Sud - Paris 11 (UP11), Université Paris Saclay (COmUE), Università degli Studi di Urbino 'Carlo Bo', Physiopathologie et traitement des maladies du foie, Université Paris-Sud - Paris 11 (UP11)-Hôpital Paul Brousse-Institut National de la Santé et de la Recherche Médicale (INSERM), CHU Necker - Enfants Malades [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Université Paris Descartes - Paris 5 (UPD5), Génie et Microbiologie des Procédés Alimentaires (GMPA), Institut National de la Recherche Agronomique (INRA)-AgroParisTech, Institut Jean-Pierre Bourgin (IJPB), Palacky University Olomouc, Interactions cellules souches-niches : physiologie, tumeurs et réparation tissulaire, Université Paris-Sud - Paris 11 (UP11)-Service de Santé des Armées-Hôpital Paul Brousse-Institut National de la Santé et de la Recherche Médicale (INSERM), YGM Consult sas, Partenaires INRAE, Laboratoire de Chimie et de Biochimie Pharmacologiques et Toxicologiques (LCBPT - UMR 8601), Université Paris Descartes - Paris 5 (UPD5)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Institut du Cancer et d'Immunogenetique (ICIG), Hospital Paul-Brousse, Villejuif, France
Jazyk: angličtina
Rok vydání: 2019
Předmět:
Zdroj: Journal of Pharmacology and Experimental Therapeutics
Journal of Pharmacology and Experimental Therapeutics, American Society for Pharmacology and Experimental Therapeutics, 2019, 369 (3), pp.489-502. ⟨10.1124/jpet.119.256537⟩
ISSN: 0022-3565
1521-0103
Popis: International audience; Methionine deprivation induces growth arrest and death of cancer cells. To eliminate L-methionine we produced, purified, and characterized the recombinant pyridoxal 59-phosphate (PLP)-dependent L-methionine g-lyase (MGL) BL929 from the cheese-ripening Brevibacterium aurantiacum. Transformation of an Escherichia coli strain with the gene BL929 from B. aurantiacum optimized for E. coli expression led to production of the MGL-BL929. Elimination of L-methionine and cytotoxicity in vitro were assessed, and methylation-sensitive epigenetics was explored for changes resulting from exposure of cancer cells to the enzyme. A bioreactor was built by encapsulation of the protein in human erythrocytes to achieve sustained elimination of L-methionine in extracellular fluids. Catalysis was limited to alpha, gamma-elimination of L-methionine and L-homocysteine. The enzyme had no activity on other sulfur-containing amino acids. Enzyme activity decreased in presence of serum albumin or plasma resulting from reduction of PLP availability. Elimination of L-methionine induced cytotoxicity on a vast panel of human cancer cell lines and spared normal cells. Exposure of colorectal carcinoma cells to the MGL-BL929 reduced methylCpG levels of hypermethylated gene promoters including that of CDKN2A, whose mRNA expression was increased, together with a decrease in global histone H3 dimethyl lysine 9. The MGL-erythrocyte bioreactor durably preserves enzyme activity in vitro and strongly eliminates L-methionine from medium.
Databáze: OpenAIRE
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