Trypanosoma cruzi: intra and interspecific genomic variability and mechanisms of cell invasion/egress

• Grant number: 16/15000-4
• Support Opportunities: Research Projects - Thematic Grants
• Start date: June 01, 2017
• End date: March 31, 2023
• Field of knowledge: Biological Sciences - Parasitology - Protozoology of Parasites
• Principal Investigator: Renato Arruda Mortara
• Grantee: Renato Arruda Mortara
• Host Institution: Escola Paulista de Medicina (EPM). Universidade Federal de São Paulo (UNIFESP). Campus São Paulo. São Paulo , SP, Brazil
• Pesquisadores principais: José Franco da Silveira Filho ; Nobuko Yoshida
• Associated researchers: Esteban Mauricio Cordero Veas ; Marjorie Mendes Marini e Souza ; Renata Torres de Souza
• Associated scholarship(s): 21/02881-0 - Characterization of components secreted by extracellular amastigotes of Trypanosoma cruzi and their role in cell invasion, BP.IC; 19/23302-9 - Characterization of components secreted by Trypanosoma cruzi extracellular amastigotes and their effect on cell invasion, BP.PD; 19/10858-9 - The role of host cell Na+/H+ exchanger in invasion by Trypanosoma cruzi metacyclic forms, BP.PD; + associated scholarships 19/08933-2 - Evaluation of STAT-1 and STAT-3 activation on coinfection modulation with two Trypanosoma cruzi strains in M1 and M2a polarized human macrophages, BP.IC; 19/07916-7 - Ionizing radiation effects and double-stranded breaks (DSBs) on the molecular karyotype of T. cruzi, BP.IC; 19/05049-4 - Construction of transgenic lines modified by CRISPR/Cas9 to identify events of aneuploidy in Trypanosoma cruzi, BP.PD; 17/24701-9 - The role of Cdc42, RhoA and Rac1 GTPases in actin cytoskeleton during HeLa cell invasion by Trypanosoma Cruzi metacyclic and tissue culture trypomastigotes, BP.MS; 17/23086-9 - Identification of aneuploidy events in Trypanosoma cruzi, BP.IC; 17/18791-5 - Mucin-like glycoproteins of Trypanosoma cruzi metacyclic forms: interaction with other surface molecules of the parasite and with the host cell, BP.IC; 17/23312-9 - Mechanisms involved in the generation of chromosome rearrangements and aneuploidy in Trypanosoma cruzi exposed to ionizing radiation, BP.DD; 17/17144-6 - Establishment of molecular karyotype and synthetic associations in Trypanosoma rangeli, Trypanosoma conorhini and Trypanosoma Cruzi, BP.IC; 17/20432-3 - The role of N-WASP, WAVE2 and cortactin in the internalization by extracellular amastigotes of Trypanosoma cruzi in professional phagocytic cells (THP-1), BP.IC; 17/19359-0 - Evaluation of STATs activation in JAK/STAT pathway in co-infection with two Trypanosoma cruzi strains on human macrophages, BP.IC; 17/20090-5 - Cell invasion by Trypanosoma cruzi study involving surface molecules gp90 and gp82 of metacyclic forms, BP.PD; 17/16526-2 - Identification of homologous and orthologous genes of the sap multigenic family in trypanosomes isolated from bats, Trypanosoma Cruzi marinkellei and Trypanosoma Cruzi bat, BP.IC; 16/17770-1 - Identification and characterization of novel actin cytoskeleton mediators during host cell invasion by the extracellular amastigote forms of Trypanosoma cruzi, BP.PD; 16/16918-5 - Study of Trypanosoma Cruzi trypomastigotes egress from infected cells, BP.PD - associated scholarships

Abstract

Infection by Trypanosoma cruzi, the etiological agent of Chagas Disease remains a major public health problem in Latin America, home of most people infected. In nonendemic regions like/such as the US, Europe, Japan, and Australia there is growing concern about the risks of transmission by blood transfusion (and organ transplant). Considering that T. cruzi taxon comprises genetically distinct populations with important genotypic and phenotypic characteristics and distinct infectivity patterns, we are faced with a major challenge of understanding the molecular basis of these (genetic and epigenetic) variabilities. We propose to investigate chromosomic polymorphisms and ploidy variations among different T. cruzi strains that could be part of the resources used by the parasite to cope with environmental stress. We will examine the effects of ionizing radiation on chromosome organization of the parasite, as well as the generation of recombinant events capable of changing karyotype and ploidy, which may contribute to the genetic diversity of T. cruzi. Karyotype and genome comparisons of different species of the Schyzotrypanum clade (T. cruzi e T. cruzi-like) that includes nonpathogenic trypanosomes from South America will provide insights into chromosome evolution. Different strategies and methodologies will be used, including the novel genomic editing with CRISP-Cas9, aiming at clarifying the mechanisms underlying cell invasion by metacyclic trypomastigotes and extracellular amastigotes, infective forms of T. cruzi that differ in the repertoire of interacting molecules. We will also investigate the role of cytoskeletal elements of the target cell as well as putative receptors for T. cruzi surface molecules and microvesicles released by the parasite, components of growing importance evidenced by studies with tissue culture trypomastigotes. Coinfections with different strains to elucidate mechanisms of intracellular growth and egress from parasitized mammalian cells will also be our object of study. Researchers who have been collaborating in a highly productive manner for several years with wide experience in Protozoology, with special focus on Molecular and Cellular Biology will carry out this multidisciplinary project. With the aid of researchers in Brazil and abroad, we aim to expand the knowledge on the topics proposed herein. (AU)

Scientific publications

(17)

(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)

WATANABE COSTA, RENATA; BATISTA, MARINA FERREIRA; MENEGHELLI, ISABELA; VIDAL, RAMON OLIVEIRA; NAJERA, CARLOS ALCIDES; MENDES, ANA CLARA; ANDRADE-LIMA, IZABELA AUGUSTA; DA SILVEIRA, JOSE FRANCO; LOPES, LUCIANO RODRIGO; FERREIRA, LUDMILA RODRIGUES PINTO; et al. Comparative Analysis of the Secretome and Interactome of Trypanosoma cruzi and Trypanosoma rangeli Reveals Species Specific Immune Response Modulating Proteins. FRONTIERS IN IMMUNOLOGY, v. 11, AUG 27 2020. (16/15000-4, 15/50315-3)

MEDINA, CAMILA MACEDO; FERREIRA, EDEN RAMALHO; BONIFACIO, BRUNO SOUZA; MORTARA, RENATO ARRUDA; BONFIM-MELO, ALEXIS. rypanosoma cruzi extracellular amastigotes engage Rac1 and Cdc42 to invade RAW macrophage. Microbes and Infection, v. 23, n. 8, SEP-OCT 2021. (17/20432-3, 16/15000-4)

ONOFRE, THIAGO SOUZA; FERREIRA RODRIGUES, JOAO PAULO; YOSHIDA, NOBUKO. Depletion of Host Cell Focal Adhesion Kinase Increases the Susceptibility to Invasion by Trypanosoma cruzi Metacyclic Forms. FRONTIERS IN CELLULAR AND INFECTION MICROBIOLOGY, v. 9, JUN 26 2019. (16/15000-4)

DE CASTRO NETO, ARTUR LEONEL; DA SILVEIRA, JOSE FRANCO; MORTARA, RENATO ARRUDA. Comparative Analysis of Virulence Mechanisms of Trypanosomatids Pathogenic to Humans. FRONTIERS IN CELLULAR AND INFECTION MICROBIOLOGY, v. 11, APR 16 2021. (19/23302-9, 16/15000-4)

OLIVEIRA, MELISSA MARTINS; BONTURI, CAMILA RAMALHO; SALU, BRUNO RAMOS; OLIVA, MARIA LUIZA VILELA; MORTARA, RENATO ARRUDA; ORIKAZA, CRISTINA MARY. Modulation of STAT-1, STAT-3, and STAT-6 activities in THP-1 derived macrophages infected with two Trypanosoma cruzi strains. FRONTIERS IN IMMUNOLOGY, v. 13, p. 14-pg., 2022-10-26. (16/15000-4, 19/08933-2)

ORIKAZA, CRISTINA M.; PESSOA, CARINA C.; PALADINO, V, FERNANDA; FLORENTINO, V, PILAR T.; BARBIERI, CLARA L.; GOTO, HIRO; RAMOS-SANCHEZ, EDUARDO MILTON; SILVEIRA, JOSE FRANCO; RABINOVITCH, MICHEL; MORTARA, RENATO A.; et al. Dual Host-Intracellular Parasite Transcriptome of Enucleated Cells Hosting Leishmania amazonensis: Control of Half-Life o Host Cell Transcripts by the Parasite. Infection and Immunity, v. 88, n. 11, NOV 2020. (14/25296-2, 16/15000-4)

TEIXEIRA, THAISE LARA; CHIURILLO, MIGUEL ANGEL; LANDER, NOELIA; RODRIGUES, CASSIANO COSTA; ONOFRE, THIAGO SOUZA; FERREIRA, EDEN RAMALHO; YONAMINE, CAMILA MIYAGUI; SANTOS, JULIA DE GOUVEIA; MORTARA, RENATO ARRUDA; DA SILVA, CLAUDIO VIEIRA; et al. Ablation of the P21 Gene of Trypanosoma cruzi Provides Evidence of P21 as a Mediator in the Control of Epimastigote and Intracellular Amastigote Replication. FRONTIERS IN CELLULAR AND INFECTION MICROBIOLOGY, v. 12, p. 11-pg., 2022-02-18. (19/05049-4, 16/15000-4)

MORETTI, NILMAR SILVIO; MORTARA, RENATO ARRUDA; SCHENKMAN, SERGIO. Trypanosoma cruzi. Trends in Parasitology, v. 36, n. 4, p. 2-pg., 2020-04-01. (15/22031-0, 16/15000-4)

BONIFACIO, BRUNO SOUZA; BONFIM-MELO, ALEXIS; MORTARA, RENATO ARRUDA; FERREIRA, EDEN RAMALHO. Successful invasion of Trypanosoma cruzi trypomastigotes is dependent on host cell actin cytoskeleton. Journal of Eukaryotic Microbiology, v. 69, n. 3, p. 10-pg., 2022-03-31. (16/16918-5, 17/24701-9, 16/17770-1, 16/15000-4)

BERNARDO, WERICA P.; SOUZA, RENATA T.; COSTA-MARTINS, ANDRE G.; FERREIRA, EDEN R.; MORTARA, RENATO A.; TEIXEIRA, MARTA M. G.; RAMIREZ, JOSE LUIS; DA SILVEIRA, JOSE F.. Genomic Organization and Generation of Genetic Variability in the RHS (Retrotransposon Hot Spot) Protein Multigene Family in Trypanosoma cruzi. GENES, v. 11, n. 9, SEP 2020. (16/15000-4, 16/16918-5)

PESSOA, CARINA CARRARO; REIS, LUIZA CAMPOS; RAMOS-SANCHEZ, EDUARDO MILTON; ORIKAZA, CRISTINA MARY; CORTEZ, CRISTIAN; DE CASTRO LEVATTI, ERICA VALADARES; BENITES BADARO, ANA CAROLINA; DA SILVA YAMAMOTO, JOYCE UMBELINO; D'ALMEIDA, VANIA; GOTO, HIRO; et al. ATP6V(0)d2 controls Leishmania parasitophorous vacuole biogenesis via cholesterol homeostasis. PLOS PATHOGENS, v. 15, n. 6, JUN 2019. (15/14205-9, 16/15000-4)

GONZATTI, MICHELANGELO BAUWELZ; PERRUD SOUSA, MARIA EDUARDA; TUNISSI, ARIANE SIMOES; MORTARA, RENATO ARRUDA; DE OLIVEIRA, ADRIANO MARIM; PEREIRA CERIZE, NATALIA NETO; KELLER, ALEXANDRE DE CASTRO. Nano spray dryer for vectorizing alpha-galactosylceramide in polymeric nanoparticles: A single step process to enhance invariant Natural Killer T lymphocyte responses. International Journal of Pharmaceutics, v. 565, p. 123-132, JUN 30 2019. (16/02189-1, 17/12799-4, 16/15000-4)

FERREIRA RODRIGUES, JOAO PAULO; ONOFRE, THIAGO SOUZA; BARBOSA, BRUNO COUTO; FERREIRA, EDEN RAMALHO; BONFIM-MELO, ALEXIS; YOSHIDA, NOBUKO. Host cell protein LAMP-2 is the receptor for Trypanosoma cruzi surface molecule gp82 that mediates invasion. Cellular Microbiology, v. 21, n. 5, MAY 2019. (16/15000-4)

LOCH, LEONARDO; ONOFRE, THIAGO SOUZA; FERREIRA RODRIGUES, JOAO PAULO; YOSHIDA, NOBUKO. Shedding of Trypanosoma cruzi Surface Molecules That Regulate Host Cell Invasion Involves Phospholipase C and Increases Upon Sterol Depletion. FRONTIERS IN CELLULAR AND INFECTION MICROBIOLOGY, v. 11, OCT 19 2021. (16/15000-4)

DE LIRA SILVA, NADJANIA SARAIVA; ORIKAZA, CRISTINA MARY; DE SANTANA, FABIANA RODRIGUES; DOS SANTOS, LUANA AGUIAR; SALU, BRUNO RAMOS; VILELA OLIVA, MARIA LUIZA; SINIGAGLIA, RITA DE CASSIA; MORTARA, RENATO ARRUDA. Interleukin-9 in Immunopathology of Trypanosoma cruzi Experimental Infection. FRONTIERS IN CELLULAR AND INFECTION MICROBIOLOGY, v. 11, p. 15-pg., 2021-10-15. (16/15000-4)

FERREIRA RODRIGUES, JOAO PAULO; LOCH, LEONARDO; ONOFRE, THIAGO SOUZA; YOSHIDA, NOBUKO. Depletion of Na+/H+ Exchanger Isoform 1 Increases the Host Cell Resistance to Trypanosoma cruzi Invasion. PATHOGENS, v. 11, n. 11, p. 13-pg., 2022-11-01. (16/15000-4)

ONOFRE, THIAGO SOUZA; FERREIRA RODRIGUES, JOAO PAULO; SHIO, MARINA TIEMI; MACEDO, SILENE; JULIANO, MARIA APARECIDA; YOSHIDA, NOBUKO. Interaction of Trypanosoma cruzi Gp82 With Host Cell LAMP2 Induces Protein Kinase C Activation and Promotes Invasion. FRONTIERS IN CELLULAR AND INFECTION MICROBIOLOGY, v. 11, MAR 12 2021. (16/15000-4)