Wanderson Duarte DaRocha

Scopus Publications

Kharon Is Crucial for Trypanosoma cruzi Morphology but Does Not Impair In Vitro Infection
Jose Luis Saenz-Garcia, Normanda Souza-Melo, Juliana Severo Miranda, Beatriz Borges, Lisandro A. Pacheco-Lugo, et al.
Pathogens, 2025
Chagas disease, caused by Trypanosoma cruzi, is a neglected tropical disease with few options for treatment and no available vaccine. Deletion mutants for live attenuated vaccines, particularly deletions of proteins related to the cytoskeleton, have been widely tested in related parasites but candidates have not been tested in T. cruzi. Kharon is one such protein, identified as being associated with the cytoskeleton in Leishmania and essential for amastigote replication. Here we investigated the T. cruzi Kharon ortholog (TcKharon) to test if it has orthologous function and thus potential in generating a live attenuated vaccine. In silico analysis predicted TcKharon to be an intrinsically disordered protein, consistent with its ortholog feature, and GFP fusion protein revealed that TcKharon is associated with the cytoskeleton of epimastigotes. CRISPR-Cas9-mediated gene disruption impaired epimastigote proliferation and cytokinesis, resulting in altered nucleus-to-kinetoplast ratios and pronounced morphological defects, particularly in the posterior cell region. Despite these abnormalities, TcKharon−/− mutants retained the ability to differentiate into metacyclic trypomastigotes and exhibited in vitro infection rates comparable to wild-type parasites. Our data show that TcKharon is crucial for cell morphology. However, in contrast to close related parasites, TcKharon is not essential for in vitro infectivity.
Trypanosoma cruzi Vps34 colocalizes with Beclin1 and plays a role in parasite invasion of the host cell by modulating the expression of a sub-group of trans-sialidases
Carlos Alcides Nájera, Mercedes Soares-Silva, Fernando Y. Maeda, Wanderson Duarte DaRocha, Isabela Meneghelli, et al.
Microbes and Infection, 2024
Lipidomic Analysis Reveals Branched-Chain and Cyclic Fatty Acids from Angomonas deanei Grown under Different Nutritional and Physiological Conditions
Arquimedes Paixão Santana-Filho, Aramís José Pereira, Letícia Adejani Laibida, Normanda Souza-Melo, Wanderson Duarte DaRocha, et al.
Molecules, 2024
Angomonas deanei belongs to Trypanosomatidae family, a family of parasites that only infect insects. It hosts a bacterial endosymbiont in a mutualistic relationship, constituting an excellent model for studying organelle origin and cellular evolution. A lipidomic approach, which allows for a comprehensive analysis of all lipids in a biological system (lipidome), is a useful tool for identifying and measuring different expression patterns of lipid classes. The present study applied GC-MS and NMR techniques, coupled with principal component analysis (PCA), in order to perform a comparative lipidomic study of wild and aposymbiotic A. deanei grown in the presence or absence of FBS. Unusual contents of branched-chain iso C17:0 and C19:0-cis-9,10 and-11,12 fatty acids were identified in A. deanei cultures, and it was interesting to note that their content slightly decreased at the log phase culture, indicating that in the latter growth stages the cell must promote the remodeling of lipid synthesis in order to maintain the fluidity of the membrane. The combination of analytical techniques used in this work allowed for the detection and characterization of lipids and relevant contributors in a variety of A. deanei growth conditions.
Trypanin Disruption Affects the Motility and Infectivity of the Protozoan Trypanosoma cruzi
Jose L. Saenz-Garcia, Beatriz S. Borges, Normanda Souza-Melo, Luiz V. Machado, Juliana S. Miranda, et al.
Frontiers in Cellular and Infection Microbiology, 2022
The flagellum of Trypanosomatids is an organelle that contributes to multiple functions, including motility, cell division, and host–pathogen interaction. Trypanin was first described in Trypanosoma brucei and is part of the dynein regulatory complex. TbTrypanin knockdown parasites showed motility defects in procyclic forms; however, silencing in bloodstream forms was lethal. Since TbTrypanin mutants show drastic phenotypic changes in mammalian stages, we decided to evaluate if the Trypanosoma cruzi ortholog plays a similar role by using the CRISPR-Cas9 system to generate null mutants. A ribonucleoprotein complex of SaCas9 and sgRNA plus donor oligonucleotide were used to edit both alleles of TcTrypanin without any selectable marker. TcTrypanin −/− epimastigotes showed a lower growth rate, partially detached flagella, normal numbers of nuclei and kinetoplasts, and motility defects such as reduced displacement and speed and increased tumbling propensity. The epimastigote mutant also showed decreased efficiency of in-vitro metacyclogenesis. Mutant parasites were able to complete the entire life cycle in vitro; however, they showed a reduction in their infection capacity compared with WT and addback cultures. Our data show that T. cruzi life cycle stages have differing sensitivities to TcTrypanin deletion. In conclusion, additional work is needed to dissect the motility components of T. cruzi and to identify essential molecules for mammalian stages.
CREditing: a tool for gene tuning in Trypanosoma cruzi
Lisandro A. Pacheco-Lugo, José L. Sáenz-García, Yirys Díaz-Olmos, Rodrigo Netto-Costa, Rodrigo S.C. Brant, et al.
International Journal for Parasitology, 2020
Targeting epimastigotes of Trypanosoma cruzi with a peptide isolated from a phage display random library
José L. Sáenz-Garcia, Isabel B. Yamanaka, Lisandro A. Pacheco-Lugo, Juliana S. Miranda, Emily S. Córneo, et al.
Experimental Parasitology, 2020
Engineering a single-chain antibody against Trypanosoma cruzi metacyclic trypomastigotes to block cell invasion
Lara Maria Kalempa Demeu, Rodrigo Jahn Soares, Juliana Severo Miranda, Lisandro A. Pacheco-Lugo, Kelin Gonçalves Oliveira, et al.
Plos One, 2019
Trypanosoma cruzi is a flagellate protozoan pathogen that causes Chagas disease. Currently there is no preventive treatment and the efficiency of the two drugs available is limited to the acute phase. Therefore, there is an unmet need for innovative tools to block transmission in endemic areas. In this study, we engineered a novel recombinant molecule able to adhere to the T. cruzi surface, termed scFv-10D8, that consists of a single-chain variable fragment (scFv) derived from mAb-10D8 that targets gp35/50. The synthetic gene encoding scFv-10D8 was cloned and fused to a 6×His tag and expressed in a prokaryotic expression system. Total periplasmic or 6xHis tag affinity-purified fractions of scFv-10D8 retained the capacity to bind to gp35/50, as shown by Western blot analyses. Pre-incubation of metacyclic trypomastigotes with scFv-10D8 showed a remarkable reduction in cell invasion capacity. Our results suggest that scFv-10D8 can be used in a paratransgenic approach to target parasites in insect vectors, avoiding dissemination of infective forms. Such advances in the development of this functional molecule will surely prompt the improvement of alternative strategies to control Chagas disease by targeting mammalian host stages.
Assessment of two CRISPR-Cas9 genome editing protocols for rapid generation of Trypanosoma cruzi gene knockout mutants
Gabriela Assis Burle-Caldas, Melissa Soares-Simões, Laiane Lemos-Pechnicki, Wanderson Duarte DaRocha, Santuza M.R. Teixeira
International Journal for Parasitology, 2018
Effective gene delivery to Trypanosoma cruzi epimastigotes through nucleofection
Lisandro Pacheco-Lugo, Yirys Díaz-Olmos, José Sáenz-García, Christian Macagnan Probst, Wanderson Duarte DaRocha
Parasitology International, 2017
Zinc finger nuclease technology: A stable tool for high efficiency transformation in bloodstream form T. brucei
Gabriela Schumann, Monica M. Kangussu-Marcolino, Nicholas Doiron, Sandro Käser, Gabriela de Assis Burle-Caldas, et al.
Molecular and Biochemical Parasitology, 2017
Editing the Trypanosoma cruzi genome with zinc finger nucleases
Gabriela Assis Burle-Caldas, Viviane Grazielle-Silva, Melissa Soares-Simões, Gabriela Schumann Burkard, Isabel Roditi, et al.
Molecular and Biochemical Parasitology, 2017
Expanding the tool box for genetic manipulation of Trypanosoma cruzi
Gabriela de Assis Burle-Caldas, Viviane Grazielle-Silva, Letícia Adejani Laibida, Wanderson Duarte DaRocha, Santuza Maria Ribeiro Teixeira
Molecular and Biochemical Parasitology, 2015
Amastin Knockdown in Leishmania braziliensis Affects Parasite-Macrophage Interaction and Results in Impaired Viability of Intracellular Amastigotes
Rita Marcia Cardoso de Paiva, Viviane Grazielle-Silva, Mariana Santos Cardoso, Brenda Naemi Nakagaki, Rondon Pessoa Mendonça-Neto, et al.
Plos Pathogens, 2015
Unveiling the Intracellular Survival Gene Kit of Trypanosomatid Parasites
Daniella Castanheira Bartholomeu, Rita Marcia Cardoso de Paiva, Tiago A. O. Mendes, Wanderson D. DaRocha, Santuza M. R. Teixeira
Plos Pathogens, 2014
Genome of the Avirulent Human-Infective Trypanosome—Trypanosoma rangeli
Patrícia Hermes Stoco, Glauber Wagner, Carlos Talavera-Lopez, Alexandra Gerber, Arnaldo Zaha, et al.
Plos Neglected Tropical Diseases, 2014
Differential ability to resist to complement lysis and invade host cells mediated by MBL in R4 and 860 strains of Trypanosoma cruzi
Ingrid Evans-Osses, Andres Mojoli, Marcia Holsbach Beltrame, Denise Endo da Costa, Wanderson Duarte DaRocha, et al.
FEBS Letters, 2014
Conditional removal of selectable markers in Trypanosoma cruzi using a site-specific recombination tool: Proof of concept
Monica M. Kangussu-Marcolino, Ana Paula Cunha, Andrea R. Avila, Jean-Paul Herman, Wanderson D. DaRocha
Molecular and Biochemical Parasitology, 2014
Identification of secreted virulence factors of Chromobacterium violaceum
Thiago Castro-Gomes, Mariana S. Cardoso, Wanderson D. DaRocha, Letícia A. Laibida, Andréa M. A. Nascimento, et al.
Journal of Microbiology, 2014
Repeat-enriched proteins are related to host cell invasion and immune evasion in parasitic protozoa
T.A.O. Mendes, F.P. Lobo, T.S. Rodrigues, G.F. Rodrigues-Luiz, W.D. daRocha, et al.
Molecular Biology and Evolution, 2013
Distinct genomic organization, mRNA expression and cellular localization of members of two amastin sub-families present in Trypanosoma cruzi
Monica Mendes Kangussu-Marcolino, Rita Márcia Cardoso de Paiva, Patrícia Rosa Araújo, Rondon Pessoa de Mendonça-Neto, Laiane Lemos, et al.
BMC Microbiology, 2013
Antigenicity and Protective Efficacy of a Leishmania Amastigote-specific Protein, Member of the Super-oxygenase Family, against Visceral Leishmaniasis
Vivian T. Martins, Miguel A. Chávez-Fumagalli, Lourena E. Costa, Adriana M. C. C. Martins, Paula S. Lage, et al.
Plos Neglected Tropical Diseases, 2013
Trypanosoma cruzi: Role of δ-Amastin on Extracellular Amastigote Cell Invasion and Differentiation
Mário C. Cruz, Normanda Souza-Melo, Claudio Vieira da Silva, Wanderson Duarte DaRocha, Diana Bahia, et al.
Plos One, 2012
Genomic comparison of the endophyte Herbaspirillum seropedicae SmR1 and the phytopathogen Herbaspirillum rubrisubalbicans M1 by suppressive subtractive hybridization and partial genome sequencing
Rose A. Monteiro, Eduardo Balsanelli, Thalita Tuleski, Helison Faoro, Leonardo M. Cruz, et al.
FEMS Microbiology Ecology, 2012
Trypanosomatid comparative genomics: Contributions to the study of parasite biology and different parasitic diseases
Santuza M. Teixeira, Rita Márcia Cardoso de Paiva, Monica M. Kangussu-Marcolino, Wanderson D. DaRocha
Genetics and Molecular Biology, 2012
Trypanosoma cruzi as an effective cancer antigen delivery vector
Caroline Junqueira, Luara I. Santos, Bruno Galvão-Filho, Santuza M. Teixeira, Flávia G. Rodrigues, et al.
Proceedings of the National Academy of Sciences of the United States of America, 2011
An essential nuclear protein in trypanosomes is a component of mrna transcription/export pathway
Mariana Serpeloni, Carolina Borsoi Moraes, João Renato Carvalho Muniz, Maria Cristina Machado Motta, Augusto Savio Peixoto Ramos, et al.
Plos One, 2011
Development of a dual reporter system to identify regulatory cis-acting elements in untranslated regions of Trypanosoma cruzi mRNAs
Patrícia R. Araújo, Gabriela A. Burle-Caldas, Rosiane A. Silva-Pereira, Daniella C. Bartholomeu, Wanderson D. daRocha, et al.
Parasitology International, 2011
Trypanosoma cruzi MSH2: Functional analyses on different parasite strains provide evidences for a role on the oxidative stress response
Priscila C. Campos, Viviane G. Silva, Carolina Furtado, Alice Machado-Silva, Wanderson D. DaRocha, et al.
Molecular and Biochemical Parasitology, 2011
Genomic organization and expression profile of the mucin-associated surface protein (masp) family of the human pathogen Trypanosoma cruzi
Daniella C. Bartholomeu, Gustavo C. Cerqueira, Ana Carolina A. Leão, Wanderson D. daRocha, Fabiano S. Pais, et al.
Nucleic Acids Research, 2009
DNA polymerase kappa from Trypanosoma cruzi localizes to the mitochondria, bypasses 8-oxoguanine lesions and performs DNA synthesis in a recombination intermediate
M. A. Rajão, D. G. Passos‐Silva, W. D. DaRocha, G. R. Franco, A. M. Macedo, et al.
Molecular Microbiology, 2009
Biochemical studies with DNA polymerase β and DNA polymerase β-PAK of Trypanosoma cruzi suggest the involvement of these proteins in mitochondrial DNA maintenance
Débora de Oliveira Lopes, Bruno Luiz Fonseca Schamber-Reis, Carlos Gustavo Regis-da-Silva, Matheus Andrade Rajão, Wanderson Duarte DaRocha, et al.
DNA Repair, 2008
Sequences involved in mRNA processing in Trypanosoma cruzi
Priscila C. Campos, Daniella C. Bartholomeu, Wanderson D. DaRocha, Gustavo C. Cerqueira, Santuza M.R. Teixeira
International Journal for Parasitology, 2008
Molecular characterization of ribonucleoproteic antigens containing repeated amino acid sequences from Trypanosoma cruzi
Fabiano S. Pais, Wanderson D. DaRocha, Ramon M. Almeida, Sophie Y. Leclercq, Marcus L. Penido, et al.
Microbes and Infection, 2008
Cell culture and animal infection with distinct Trypanosoma cruzi strains expressing red and green fluorescent proteins
S.F. Pires, W.D. DaRocha, J.M. Freitas, L.A. Oliveira, G.T. Kitten, et al.
International Journal for Parasitology, 2008
Sequence diversity and evolution of multigene families in Trypanosoma cruzi
Gustavo C. Cerqueira, Daniella C. Bartholomeu, Wanderson D. DaRocha, Lihua Hou, Danielle M. Freitas-Silva, et al.
Molecular and Biochemical Parasitology, 2008
Genotyping of Chlamydia trachomatis from endocervical specimens in Brazil
Haleta E. Lima, Milena B. Oliveira, Brenda G. Valente, Daniela A. F. Afonso, Wanderson D. DaRocha, et al.
Sexually Transmitted Diseases, 2007
Characterization of a Trypanosoma cruzi antigen with homology to intracellular mammalian lectins
C.D.S. Macêdo, W.D. DaRocha, R.A. Mortara, M.A.M. Prado, S.M.R. Teixeira
International Journal for Parasitology, 2006
Characterization of the Trypanosoma cruzi Rad51 gene and its role in recombination events associated with the parasite resistance to ionizing radiation
Carlos Gustavo Regis-da-Silva, Jorge Marcelo Freitas, Danielle Gomes Passos-Silva, Carolina Furtado, Luiz Augusto-Pinto, et al.
Molecular and Biochemical Parasitology, 2006
Analysis of expressed sequence tags from Trypanosoma cruzi amastigotes
Gustavo C Cerqueira, Wanderson D DaRocha, Priscila C Campos, Cláudia S Zouain, Santuza MR Teixeira
Memorias do Instituto Oswaldo Cruz, 2005
Expression of exogenous genes in Trypanosoma cruzi: Improving vectors and electroporation protocols
Wanderson D. DaRocha, Rosiane A. Silva, Daniella C. Bartholomeu, Simone F. Pires, Jorge M. Freitas, et al.
Parasitology Research, 2004
Tests of cytoplasmic RNA interference (RNAi) and construction of a tetracycline-inducible T7 promoter system in Trypanosoma cruzi
Wanderson D. DaRocha, Keiko Otsu, Santuza M.R. Teixeira, John E. Donelson
Molecular and Biochemical Parasitology, 2004
Control of gene expression and genetic manipulation in the Trypanosomatidae
Genetics and Molecular Research, 2003
Characterization of cDNA clones encoding ribonucleoprotein antigens expressed in Trypanosoma cruzi amastigotes
Wanderson D. DaRocha, Daniella C. Bartholomeu, Camila D. Macêdo, Fátima M. Horta, Edécio Cunha-Neto, et al.
Parasitology Research, 2002

Wanderson Duarte DaRocha

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Scopus Publications