Amino acid metabolism in Trypanosoma cruzi: a toolbox to survive in hostile environments

• Grant number: 21/12938-0
• Support Opportunities: Research Projects - Thematic Grants
• Start date: April 01, 2023
• End date: March 31, 2028
• 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
• Principal investigators: Otavio Henrique Thiemann
• Associated researchers: Achim Schnaufer ; Brian Alejandro Suárez Mantilla ; Claudio Alejandro Pereira ; Frederique Bringaud ; Hendrik Pieter de Koning ; Igor Correia de Almeida ; Janaina de Freitas Nascimento ; John Morrison Kelly ; Julia Pinheiro Chagas da Cunha ; Maria Carolina Quartim Barbosa Elias Sabbaga ; Martin Taylor ; Michael Barrett ; Paul Michels ; Richard Burchmore ; Roberto Docampo
• Associated research grant(s): 25/17565-8 - 6th Mini Symposium: Implications of survival and death mechanisms for the interactions of pathogenic protozoa with their hos, AO.R; 24/14603-3 - Discovering and developing novel therapeutics for Chagas disease, AV.EXT; 24/14375-0 - The role of alternative oxidases in Trypanosomatidae, AV.EXT; + associated grants 23/15330-8 - Influence of Enzyme Deacetylation on the Metabolism of Trypanosoma cruzi, AV.EXT; 23/08572-5 - Evaluation of original, and bioinformatically modifiable, marine antimicrobial peptides as trypanocidal agents., AV.EXT; 23/06066-5 - 3D cultures to understand host-parasite interactions., AV.EXT; 23/08204-6 - Building up computational models of the Trypanosoma cruzi metabolism, AV.EXT - associated grants
• Associated scholarship(s): 24/22343-1 - The Role of Carbamoyl Phosphate Synthetase (EC 6.3.5.5) in L-Glutamine and L-Glutamate Pathway Metabolism in Trypanosoma cruzi, BP.DR; 23/15602-8 - Structural study of the enzymes P5C, P5CS, P5CR, P5CDH and PRODH of Proline metabolism in Trypanosoma cruzi as targets for the discovery of drugs against Chagas Disease, BP.DR; 24/06474-9 - Characterization and biological role of the catabolism of Cystein in Trypanosoma cruzi., BP.PD; + associated scholarships 24/07601-4 - Maintenance of strains and lineages of Trypanosoma cruzi, Trypanosoma brucei and Leishmania amazonensis in in vitro culture, BP.TT; 23/11104-3 - Functional characterization of the GF6PA and GNA genes of Trypanosoma cruzi and the relevance of glutamine for parasite biology, BP.PD; 23/07306-0 - Development and validation of the genome-scale metabolic model of Trypanosoma cruzi: comparative in silico analysis of the metabolic flux of epimastigotes, trypomastigotes and amastigotes, BP.PD; 23/16035-0 - Functional Characterization of NFS, ISD11, and MTU1 Proteins: involvement in Mitochondrial Biology in Trypanosoma cruzi., BP.DD; 22/16078-8 - Evaluation of the Succinyl-CoA Synthetase/Acetate:Succinate CoA Transferase (SCS/ASCT) cycle in the biology of Trypanosoma cruzi: contribution of amino acids in the production of mitochondrial ATP at the substrate level, bioenergetics and lipid synthesis, BP.PD; 23/07669-5 - The metabolic connection between Methionine and Cysteine: a relevant aspect of the sulfur metabolism in Trypanosoma cruzi, BP.DD; 22/09116-0 - Study of the metabolic flux to analyze the relationships between biochemical pathways of amino acids and biological processes of Trypanosoma cruzi, BP.PD; 22/14959-7 - The relevance of sulphur amino acids and their metabolism in the biology of Trypanosoma cruzi, BP.PD; 22/16258-6 - Potential of ”1- Pyrroline-5-carboxylate synthetase from Trypanosoma cruzi as a therapeutic target, BP.PD; 23/06791-1 - Characterization and biological role of the Glutamine-dependent Asparagine Synthase of Trypanosoma cruzi, BP.DD - associated scholarships

Abstract

Many studies including several of our group have described the relevance of the amino acids in the biology trypanosomatids, focusing on the group of the so-called TriTryps (which includes Trypanosoma cruzi, Trypanosoma brucei, and pathogenic species of the genus Leishmania). The TriTryps use amino acids for many other functions besides protein synthesis, such as cell differentiation, 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, including osmotic, thermal, oxidative and nutritional. The present project, a continuation of our ongoing work, aims to continue the detailed characterization of the enzymatic and non-enzymatic steps that make up these metabolic pathways and interconnect apparently compartmentalized pathways. To these aims we propose: (1) To complete the construction of a metabolic chart for the amino acids with focus on those having a role in the bioenergetic of T. cruzi. (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´ activities among different stages by targeted metabolomics. (5) To develop knock-out lineages using 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

(9)

(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)

NASCIMENTO, JANAINA DE FREITAS; BARISON, MARIA JULIA; MONTANARO, GABRIELA TORRES; MARCHESE, LETICIA; SOUZA, RODOLPHO ORNITZ OLIVEIRA; SILVA, LETICIA SOPHIA; GUARNIERI, ALESSANDRA APARECIDA; SILBER, ARIEL MARIANO. Knocking out histidine ammonia-lyase by using CRISPR-Cas9 abolishes histidine role in the bioenergetics and the life cycle of Trypanosoma cruzi. MICROBIAL CELL, v. 12, n. 1, p. 16-pg., 2025-01-01. (21/12938-0)

RAPADO, L. N.; NASCIMENTO, J. F.; MANCHOLA, N. C.; DAMASCENO, F. S.; ACHJIAN, R. W.; SILBER, A. M.. The branched chain amino acids (BCAAs) modulate the development of the intra-cellular stages of Trypanosoma cruzi. Experimental Parasitology, v. 255, p. 7-pg., 2023-11-19. (21/12938-0)

NASCIMENTO, JANAINA DE FREITAS; DAMASCENO, FLAVIA SILVA; MARSICCOBETRE, SABRINA; VITORINO, FRANCISCA NATALIA DE LUNA; ACHJIAN, RENAN WEEGE; DA CUNHA, JULIA PINHEIRO CHAGAS; SILBER, ARIEL MARIANO. Branched-chain amino acids modulate the proteomic profile of Trypanosoma cruzi metacyclogenesis induced by proline. PLoS Neglected Tropical Diseases, v. 18, n. 10, p. 25-pg., 2024-10-01. (18/15553-9, 21/12938-0)

MENEZES, A. P. J.; SILBER, A. M.; ELIAS, M. C.; DA CUNHA, J. P. C.. Trypanosoma cruzi cell cycle progression exhibits minimal variation in histone PTMs with unique histone H4 acetylation pattern. JOURNAL OF PROTEOMICS, v. 315, p. 9-pg., 2025-02-28. (13/07467-1, 21/12938-0, 20/00694-6, 24/14470-3, 24/02275-1, 19/21354-1, 18/14432-3, 18/15553-9)

SOWERBY, KATE; FREITAG-POHL, STEFANIE; MURILLO, ANA MILENA; SILBER, ARIEL MARIANO; POHL, EHMKE. Cysteine synthase: multiple structures of a key enzyme in cysteine synthesis and a potential drug target for Chagas disease and leishmaniasis. ACTA CRYSTALLOGRAPHICA SECTION D-STRUCTURAL BIOLOGY, v. 79, p. 13-pg., 2023-06-01. (21/12938-0, 17/16553-0)

DOS SANTOS, DENIS AMILTON; SOUZA, HIGO FERNANDO SANTOS; SILBER, ARIEL M.; CAMPOS BRASIL DE SOUZA, TATIANA DE ARRUDA; AVILA, ANDREA RODRIGUES. Protein kinases on carbon metabolism: potential targets for alternative chemotherapies against toxoplasmosis. FRONTIERS IN CELLULAR AND INFECTION MICROBIOLOGY, v. 13, p. 14-pg., 2023-05-23. (21/12938-0, 18/14432-3)

NASCIMENTO, JANAINA F.; SOUZA, RODOLPHO O. O.; ALENCAR, MAYKE B.; MARSICCOBETRE, SABRINA; MURILLO, ANA M.; DAMASCENO, FLAVIA S.; GIRARD, RICHARD B. M. M.; MARCHESE, LETICIA; LUEVANO-MARTINEZ, LUIS A.; ACHJIAN, RENAN W.; et al. How much (ATP) does it cost to build a trypanosome? A theoretical study on the quantity of ATP needed to maintain and duplicate a bloodstream-form Trypanosomabrucei cell. PLOS PATHOGENS, v. 19, n. 7, p. 35-pg., 2023-07-01. (21/12938-0, 18/14432-3)

ALENCAR, MAYKE B.; MARSICCOBETRE, SABRINA; MENGARDA, ANA C.; BALLARI, MARIA SOL; SILBER, ARIEL M.. Energy metabolism in Trypanosoma cruzi: the validated and putative bioenergetic and carbon sources. MBIO, v. 16, n. 6, p. 22-pg., 2025-05-20. (21/12938-0)

CARDINALI, CAMILA A. E. F.; MARTINS, YANDARA A.; MORAES, RUAN C. M.; COSTA, ANDRESSA P.; ALENCAR, MAYKE B.; SILBER, ARIEL M.; TORRAO, ANDREA S.. Exploring the Therapeutic Potential of Benfotiamine in a Sporadic Alzheimer's-Like Disease Rat Model: Insights into Insulin Signaling and Cognitive function. ACS Chemical Neuroscience, v. 15, n. 16, p. 13-pg., 2024-07-15. (17/10801-1, 21/12620-0, 21/12938-0)