The biological role of amino acids and their metabolites in Trypanosoma cruzi

• Grant number: 16/06034-2
• Support Opportunities: Research Projects - Thematic Grants
• Start date: December 01, 2016
• End date: September 30, 2022
• Field of knowledge: Biological Sciences - Parasitology - Protozoology of Parasites
• Principal Investigator: Ariel Mariano Silber
• Grantee: Ariel Mariano Silber
• Host Institution: Instituto de Ciências Biomédicas (ICB). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• City of the host institution: São Paulo
• Associated researchers: Aníbal Eugênio Vercesi ; Claudio Alejandro Pereira ; Cristina Nowicki ; Frederique Bringaud ; Igor Correia de Almeida ; Julia Pinheiro Chagas da Cunha ; María Belén Cassera ; Maria Carolina Quartim Barbosa Elias Sabbaga ; Mauro Javier Cortez Véliz ; Michael Barrett ; Michael Duszenko ; Paul Michels ; Roberto Docampo
• Associated research grant(s): 19/17695-8 - Metabolic control analysis of the proline oxidation and biosynthesis in Trypanosoma Cruzi, AV.EXT; 18/14799-4 - Development of platforms to study transmembrane proteins - towards medical, agricultural and biotechnological benefits, AP.R SPRINT; 18/09634-6 - Obtainment and functional evaluation of Trypanosoma cruzi mitochondrial complex v (F1FO-H+/ATPase), AV.EXT; 17/16553-0 - The role of the mitochondrial and glycosomal isoforms of fumarate reductase of Trypanosoma Cruzi, AV.EXT; 17/16554-6 - The role of the mitochondrial and glycosomal isoforms of fumarate reductase of Trypanosoma cruzi, AV.EXT
• Associated scholarship(s): 20/16569-6 - Understanding T. cruzi His degradation pathway using synthetic biology, BP.DD; 19/26106-6 - The relevance of the regulation of the enzymatic complex branched-chain alfa-keto-acid dehydrogenase (BCKDH) in the Cell Biology and bioenergetics of Trypanosoma cruzi, BP.PD; 20/04482-3 - Metabolic control analysis of the oxidation and biosynthesis of proline in Trypanosoma cruzi, BP.DD; + associated scholarships 19/13477-6 - Biological role of the his catabolism pathway in trypanosomatids, BP.IC; 19/09832-5 - Characterization and biological function of the asparagine synthetases in Trypanosoma cruzi, BP.DD; 19/01227-5 - Biological role of the intermediary enzymes of the catabolism of Branched Chain Amino Acids in Trypanosoma cruzi., BP.PD; 18/12660-9 - The metabolic connection between metionine and cysteine: a relevant aspect of the S metabolism in T. cruzi, BP.DD; 17/14334-9 - The relevance of the enzymes isovaleril-CoA dehydrogenase (1.3.8.4) and Enoil-CoA hidratase (4.2.1.17) in the metabolism of Leucine in Trypanosoma cruzi., BP.DR; 17/13887-4 - Biochemical and functiopnal characterization of the enzymes serine acetyl transferase and cysteine synthase of Trypanosoma Cruzi, BP.DR; 17/04808-3 - The relevance of the metabolism of glutamine in the cell biology and bioenergetics of Trypanosoma cruzi and Trypanosoma brucei, BP.PD; 17/12320-0 - Characterization and biological role of histidine catabolism in Trypanosoma cruzi, BP.PD - associated scholarships

Abstract

Several studies have described the relevance of the amino acids in the biology of several protozoa parasites such as Trypanosoma cruzi, the etiological agent of Chagas disease. Besides their role in protein synthesis and energy metabolism, amino acids are also involved in different biological processes in the parasite. Among them, it is worth mentioning the differentiation between stages, the energetic support to the host-cell invasion, the establishment of the infection and the resistance to different stresses the parasite faces during its life cycle, such as osmotic, thermal, oxidative and nutritional. Until now, different roles of several amino acids were evaluated individually, but an integrated and systematic view of the role of main amino acid metabolic pathways and their interactions is still lacking. Taking into account this gap, this project aims: (1) To develop a metabolic chart for the amino acids with focus on that having a role in the bioenergetic of T. cruzi, including the identification of auxothrophies for these metabolites. (2) To evaluate their intervention on: i) the induction of ATP synthesis, production of H2O2, respiratory burst and mitochondrial membrane potential, ii) differentiation, iii) support of the host-cells invasion, iv) resistance to osmotic, oxidative, thermal, nutritional, acidic stress as well as toxicity due to NH4+ accumulation, v) the modulation of cell cycle, autophagy and the programmed cell-death triggered by metabolic stress. (3) To evaluate the roles of enzymes participating of the metabolism of those amino acids having activities described in (2), as well as the effect of their chemical inhibition or overexpression, on different aspects of the biology of the parasite mentioned in (2). (4) To perform a comparative study of the different metabolic pathways activity among different stages by targeted metabolomics. (5) To develop knock-out lineages by CRISPR/Cas9 for the most relevant enzymes among those of the amino acids metabolism for different aspects of the biology of the parasite mentioned in (2) to evaluate their possible effects related to their therapeutic potential. To evaluate by metabolomics the effect of the gene suppression or the treatment of inhibitors on the metabolic profile of the cells. (6) To evaluate the impact of the amino acids metabolism on the management of the excess of NH4+ and the production of amino sugars and glycoconjugates. (AU)

Scientific publications

(33)

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

FARGNOLI, LUCIA; PANOZZO-ZENERE, ESTEBAN A.; PAGURA, LUCAS; BARISON, MARIA JULIA; CRICCO, JULIA A.; SILBER, ARIEL M.; LABADIE, GUILLERMO R.. Targeting L-Proline Uptake as New Strategy for Anti-chagas Drug Development. FRONTIERS IN CHEMISTRY, v. 8, AUG 25 2020. (16/06034-2)

BARBOSA, ROSICLER L.; CHAGAS DA CUNHA, JULIA PINHEIRO; MENEZES, ARTHUR T.; MELO, RAISSA DE F. P.; ELIAS, MARIA CAROLINA; SILBER, ARIEL M.; COLTRI, PATRICIA P.. Proteomic analysis of Trypanosoma cruzi spliceosome complex. JOURNAL OF PROTEOMICS, v. 223, JUL 15 2020. (17/06994-9, 17/16553-0, 16/06034-2)

DAMASCENO, FLAVIA S.; BARISON, MARIA JULIA; CRISPIM, MARCELL; SOUZA, RODOLPHO O. O.; MARCHESE, LETICIA; SILBER, ARIEL M.. L-Glutamine uptake is developmentally regulated and is involved in metacyclogenesis in Trypanosoma cruzi. Molecular and Biochemical Parasitology, v. 224, p. 17-25, SEP 15 2018. (16/06034-2, 17/16553-0)

MARCHESE, LETICIA; NASCIMENTO, JANAINA DE FREITAS; DAMASCENO, FLAVIA SILVA; BRINGAUD, FREDERIC; MICHELS, PAUL A. M.; SILBER, ARIEL MARIANO. The Uptake and Metabolism of Amino Acids, and Their Unique Role in the Biology of Pathogenic Trypanosomatids. PATHOGENS, v. 7, n. 2, JUN 2018. (16/06034-2, 17/16553-0)

AVILA, CARLA CRISTI; MULE, SIMON NGAO; ROSA-FERNANDES, LIVIA; VINER, ROSA; BARISON, MARIA JULIA; COSTA-MARTINS, ANDRE GUILLHERME; DE OLIVEIRA, GILBERTO SANTOS; GERALDES TEIXEIRA, MARTA MARIA; FARIAS MARINHO, CLAUDIO ROMERO; SILBER, ARIEL MARIANO; et al. Proteome-Wide Analysis of Trypanosoma cruzi Exponential and Stationary Growth Phases Reveals a Subcellular Compartment-Specific Regulation. GENES, v. 9, n. 8, AUG 2018. (16/06034-2, 14/06863-3)

CUEVAS-HERNANDEZ, I, ROBERTO; GIRARD, RICHARD M. B. M.; MARTINEZ-CERON, SARAI; SANTOS DA SILVA, MARCELO; ELIAS, MARIA CAROLINA; CRISPIM, MARCELL; TRUJILLO-FERRARA, JOSE G.; SILBER, ARIEL MARIANO. A Fluorinated Phenylbenzothiazole Arrests the Trypanosoma cruzi Cell Cycle and Diminishes the Infection of Mammalian Host Cells. Antimicrobial Agents and Chemotherapy, v. 64, n. 2, FEB 2020. (16/06034-2)

DOS REIS, LARISSA MENEZES; ADAMOSKI, DOUGLAS; OLIVEIRA SOUZA, RODOLPHO ORNITZ; RODRIGUES ASCENCAO, CAROLLINE FERNANDA; SOUSA DE OLIVEIRA, KRISHINA RATNA; CORREA-DA-SILVA, FELIPE; DE SA PATRONI, FABIO MALTA; DIAS, MARILIA MEIRA; CONSONNI, SILVIO ROBERTO; MENDES DE MORAES-VIEIRA, PEDRO MANOEL; et al. Dual inhibition of glutaminase and carnitine palmitoyltransferase decreases growth and migration of glutaminase inhibition-resistant triple-negative breast cancer cells. Journal of Biological Chemistry, v. 294, n. 24, p. 9342-9357, JUN 14 2019. (15/15626-8, 14/17820-3, 15/25832-4, 14/18061-9, 15/26059-7, 16/06034-2, 17/06225-5, 13/23510-4, 14/06512-6, 14/15968-3)

KRISHNA NAGAMPALLI, RAGHAVENDRA SASHI; NECIOSUP QUESNAY, JOSE EDWIN; ADAMOSKI, DOUGLAS; ISLAM, ZEYAUL; BIRCH, JAMES; SEBINELLI, HEITOR GOBBI; BRUNO MOREIRA GIRARD, RICHARD MARCEL; RODRIGUES ASCENCAO, CAROLLINE FERNANDA; FALA, ANGELA MARIA; PAULETTI, BIANCA ALVES; et al. Human mitochondrial pyruvate carrier 2 as an autonomous membrane transporter. SCIENTIFIC REPORTS, v. 8, FEB 22 2018. (17/02391-8, 14/20673-2, 16/06034-2, 15/02734-7, 15/25832-4, 14/17820-3, 14/12663-7, 14/06954-9)

BARISON, MARIA JULIA; RAPADO, LUDMILA NAKAMURA; MERINO, EMILIO F.; FURUSHO PRAL, ELIZABETH MIEKO; MANTILLA, BRIAN SUAREZ; MARCHESE, LETICIA; NOWICKI, CRISTINA; SILBER, ARIEL MARIANO; CASSERA, MARIA BELEN. Metabolomic profiling reveals a finely tuned, starvation-induced metabolic switch in Trypanosoma cruzi epimastigotes. Journal of Biological Chemistry, v. 292, n. 21, p. 8964-8977, MAY 26 2017. (16/06034-2, 13/18970-6)

CRISPIM, MARCELL; DAMASCENO, FLAVIA SILVA; HERNANDEZ, AGUSTINO; BARISON, MARIA JULIA; SAUTER, ISMAEL PRETTO; PAVANI, RAPHAEL SOUZA; MOURA, ALEXANDRE SANTOS; FURUSHO PRAL, ELIZABETH MIEKO; CORTEZ, MAURO; ELIAS, MARIA CAROLINA; et al. The glutamine synthetase of Trypanosoma cruzi is required for its resistance to ammonium accumulation and evasion of the parasitophorous vacuole during host-cell infection. PLoS Neglected Tropical Diseases, v. 12, n. 1, JAN 2018. (13/07467-1, 16/06034-2, 15/10580-0, 14/10443-0, 16/50050-2)

MARCHESE, LETICIA; OLAVARRIA, KAREL; SUAREZ MANTILLA, BRIAN; CRISTI AVILA, CARLA; OLIVEIRA SOUZA, RODOLPHO ORNITIZ; SILVA DAMASCENO, FLAVIA; CAROLINA ELIAS, MARIA; MARIANO SILBER, ARIEL. Trypanosoma cruzi synthesizes proline via a Delta(1)-pyrroline-5-carboxylate reductase whose activity is fine-tuned by NADPH cytosolic pools. Biochemical Journal, v. 477, n. 10, p. 1827-1845, MAY 2020. (16/06034-2)

SANTOS SOUZA, HIGO FERNANDO; ROCHA, SANDRA CARLA; DAMASCENO, FLAVIA SILVA; RAPADO, LUDMILA NAKAMURA; FURUSHO PRAL, ELISABETH MIEKO; FARIAS MARINHO, CLAUDIO ROMERO; SILBER, ARIEL MARIANO. The effect of memantine, an antagonist of the NMDA glutamate receptor, in in vitro and in vivo infections by Trypanosoma cruzi. PLoS Neglected Tropical Diseases, v. 13, n. 9, SEP 2019. (16/06034-2)

FARGNOLI, LUCIA; PANOZZO-ZENERE, ESTEBAN A.; PAGURA, LUCAS; BARISON, MARIA JULIA; CRICCO, JULIA A.; SILBER, ARIEL M.; LABADIE, GUILLERMO R.. Targeting L-Proline Uptake as New Strategy for Anti-chagas Drug Development. RONTIERS IN CHEMISTR, v. 8, p. 15-pg., 2020-08-25. (16/06034-2)

DE PAIVA, FERNANDA C. R.; CHAN, KAREN; SAMBORSKYY, MARKIYAN; SILBER, ARIEL M.; LEADLAY, PETER F.; DIAS, MARCIO V. B.. The crystal structure of AjiA1 reveals a novel structural motion mechanism in the adenylate-forming enzyme family. ACTA CRYSTALLOGRAPHICA SECTION D-STRUCTURAL BIOLOGY, v. 76, n. 12, p. 1201-1210, DEC 1 2020. (16/06034-2, 18/00351-1)

CUEVAS-HERNANDEZ, ROBERTO I.; GIRARD, RICHARD M. B. M.; KRSTULOVIC, LUKA; BAJIC, MIROSLAV; SILBER, ARIEL MARIANO. n aromatic imidazoline derived from chloroquinoline triggers cell cycle arrest and inhibits with high selectivity the Trypanosoma cruzi mammalian host-cells infectio. PLoS Neglected Tropical Diseases, v. 15, n. 11, NOV 2021. (16/06034-2)

LUEVANO-MARTINEZ, LUIS ALBERTO; GIRARD, RICHARD M. B. M.; ALENCAR, MAYKE BEZERRA; SILBER, ARIEL MARIANO. ATP regulates the activity of an alternative oxidase in Trypanosoma brucei. FEBS Letters, v. 594, n. 13, p. 2150-2158, JUL 2020. (16/06034-2, 16/12999-0)

ROCHA, SANDRA CARLA; ROSA PEREZ, ANA; BELOSCAR, JUAN; BOTTASSO, OSCAR; SILBER, ARIEL MARIANO. Diminished Prolinemia in Chronic Chagasic Patients: A New Clue for Disease Pathology?. Molecules, v. 24, n. 17, SEP 1 2019. (16/06034-2, 17/16553-0)

ROXANA GUTIERREZ-ARMIJOS, L.; SUSSMANN, RODRIGO A. C.; SILBER, ARIEL M.; CORTEZ, MAURO; HERNANDEZ, AGUSTIN. Abnormal sterol-induced cell wall glucan deficiency in yeast is due to impaired glucan synthase transport to the plasma membrane. Biochemical Journal, v. 477, n. 24, p. 4729-4744, DEC 2020. (14/10443-0, 16/06034-2, 12/24105-3)

OLIVEIRA SOUZA, RODOLPHO ORNITZ; CRISPIM, MARCELL; SILBER, ARIEL MARIANO; DAMASCENO, FLAVIA SILVA. Glutamine Analogues Impair Cell Proliferation, the Intracellular Cycle and Metacyclogenesis in Trypanosoma cruzi. Molecules, v. 25, n. 7, APR 2020. (16/06034-2)

DE ARAUJO, CHRISTIANE BEZERRA; DE LIMA, LOYZE PAOLA; CALDERANO, SIMONE GUEDES; DAMASCENO, FLAVIA SILVA; SILBER, ARIEL M.; ELIAS, MARIA CAROLINA. Pep5, a Fragment of Cyclin D2, Shows Antiparasitic Effects in Different Stages of the Trypanosoma cruzi Life Cycle and Blocks Parasite Infectivity. Antimicrobial Agents and Chemotherapy, v. 63, n. 5, MAY 2019. (13/07467-1, 16/06034-2, 17/16553-0, 16/50050-2, 14/13375-5)

SHIRATSUBAKI, ISABEL S.; FANG, XIN; SOUZA, RODOLPHO O. O.; PALSSON, BERNHARD O.; SILBER, ARIEL M.; SIQUEIRA-NETO, JAIR L.. Genome-scale metabolic models highlight stage-specific differences in essential metabolic pathways in Trypanosoma cruzi. PLoS Neglected Tropical Diseases, v. 14, n. 10, OCT 2020. (16/06034-2, 18/14432-3)

SANTOS SOUZA, HIGO FERNANDO; REAL, DANIEL; LEONARDI, DARIO; ROCHA, SANDRA CARLA; ALONSO, VICTORIA; SERRA, ESTEBAN; SILBER, ARIEL MARIANO; JAVIER SALOMON, CLAUDIO. Development andin vitro/in vivoevaluation of a novel benznidazole liquid dosage form using a quality-by-design approach. TROPICAL MEDICINE & INTERNATIONAL HEALTH, v. 22, n. 12, p. 1514-1522, DEC 2017. (16/06034-2, 13/18970-6)

BOREIKO, SHEILA; SILVA, MARCIO; MELO, RAISSA DE F. P.; SILBER, ARIEL M.; IULEK, JORGE. Structure of Urocanate Hydratase from the protozoan Trypanosoma cruzi. International Journal of Biological Macromolecules, v. 146, p. 716-724, MAR 1 2020. (08/57910-0, 16/06034-2)

MELO, RAISSA DE FATIMA PIMENTEL; GUARNERI, ALESSANDRA APARECIDA; SILBER, ARIEL MARIANO. The Influence of Environmental Cues on the Development of Trypanosoma cruzi in Triatominae Vector. FRONTIERS IN CELLULAR AND INFECTION MICROBIOLOGY, v. 10, FEB 21 2020. (16/06034-2)

MANTILLA, BRIAN S.; MARCHESE, LETICIA; CASAS-SANCHEZ, AITOR; DYER, NAOMI A.; EJEH, NICHOLAS; BIRAN, MARC; BRINGAUD, FREDERIC; LEHANE, MICHAEL J.; ACOSTA-SERRANO, ALVARO; SILBER, ARIEL M.. Proline Metabolism is Essential for Trypanosoma brucei brucei Survival in the Tsetse Vector. PLOS PATHOGENS, v. 13, n. 1, JAN 2017. (11/22697-8, 16/06034-2)

FERRARINI, MARIANA GALVAO; NISIMURA, LINDICE MITIE; BRUNO MOREIRA GIRARD, RICHARD MARCEL; ALENCAR, MAYKE BEZERRA; ISHIKAWA FRAGOSO, MARIANA SAYURI; ARAUJO-SILVA, CARLLA ASSIS; VEIGA, ALAN DE ALMEIDA; RESSETTI ABUD, ANA PAULA; NARDELLI, SHEILA CRISTINA; VOMMARO, ROSSIANE C.; et al. Dichloroacetate and Pyruvate Metabolism: Pyruvate Dehydrogenase Kinases as Targets Worth Investigating for Effective Therapy of Toxoplasmosis. MSPHERE, v. 6, n. 1, JAN-FEB 2021. (16/06034-2)

GIRARD, RICHARD M. B. M.; CRISPIM, MARCELL; ALENCAR, MAYKE BEZERRA; SILBER, ARIEL MARIANO. Uptake of L-Alanine and Its Distinct Roles in the Bioenergetics of Trypanosoma cruzi. MSPHERE, v. 3, n. 4, JUL-AUG 2018. (16/06034-2, 17/16553-0)

MANCHOLA, NUBIA CAROLINA; SILBER, ARIEL MARIANO; NOWICKI, CRISTINA. The Non-Canonical Substrates of Trypanosoma cruzi Tyrosine and Aspartate Aminotransferases: Branched-Chain Amino Acids. Journal of Eukaryotic Microbiology, v. 65, n. 1, p. 70-76, JAN-FEB 2018. (13/18970-6, 16/06034-2)

MANTILLA, BRIAN S.; PAES-VIEIRA, LISVANE; DIAS, FELIPE DE ALMEIDA; CALDERANO, SIMONE G.; ELIAS, MARIA CAROLINA; COSENTINO-GOMES, DANIELA; OLIVEIRA, PEDRO L.; MEYER-FERNANDES, JOSE ROBERTO; SILBER, ARIEL M.. Higher expression of proline dehydrogenase altered mitochondrial function and increased Trypanosoma cruzi differentiation in vitro and in the insect vector. Biochemical Journal, v. 478, n. 21, p. 3891-3903, NOV 2021. (16/06034-2, 18/14432-3)

MICHELS, PAUL A. M.; VILLAFRAZ, ORIANA; PINEDA, ERIKA; ALENCAR, MAYKE B.; CACERES, ANA J.; SILBER, ARIEL M.; BRINGAUD, FREDERIC. Carbohydrate metabolism in trypanosomatids: New insights revealing novel complexity, diversity and species-unique features. Experimental Parasitology, v. 224, MAY 2021. (18/14432-3, 16/06034-2)

SOUZA, RODOLPHO ORNITZ OLIVEIRA; DAMASCENO, FLAVIA SILVA; MARSICCOBETRE, SABRINA; BIRAN, MARC; MURATA, GILSON; CURI, RUI; BRINGAUD, FREDERIC; SILBER, ARIEL MARIANO. Fatty acid oxidation participates in resistance to nutrient-depleted environments in the insect stages of Trypanosoma cruzi. PLOS PATHOGENS, v. 17, n. 4, APR 2021. (18/14432-3, 16/06034-2)

VILLAFRAZ, ORIANA; BIRAN, MARC; PINEDA, ERIKA; PLAZOLLES, NICOLAS; CAHOREAU, EDERN; ORNITZ OLIVEIRA SOUZA, RODOLPHO; THONNUS, MAGALI; ALLMANN, STEFAN; TETAUD, EMMANUEL; RIVIERE, LOIC; et al. Procyclic trypanosomes recycle glucose catabolites and TCA cycle intermediates to stimulate growth in the presence of physiological amounts of proline. PLOS PATHOGENS, v. 17, n. 3, MAR 2021. (18/14432-3, 16/06034-2)

DE PAIVA, FERNANDA C. R.; CHAN, KAREN; SAMBORSKYY, MARKIYAN; SILBER, ARIEL M.; LEADLAY, PETER F.; DIAS, MARCIO V. B.. The crystal structure of AjiA1 reveals a novel structural motion mechanism in the adenylate-forming enzyme family. ACTA CRYSTALLOGRAPHICA SECTION D-STRUCTURAL BIOLOGY, v. 76, p. 10-pg., 2020-12-01. (18/00351-1, 16/06034-2)