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A Novel High-Content Screening-Based Method for Anti-Trypanosoma cruzi Drug Discovery Using Human-Induced Pluripotent Stem Cell-Derived Cardiomyocytes

Chagas disease is caused by Trypanosoma cruzi infection and remains a relevant cause of chronic heart failure in Latin America. The pharmacological arsenal for Chagas disease is limited, and the available anti-T. cruzi drugs are not effective when administered during the chronic phase. Cardiomyocyte...

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Autores principales: Portella, Diogo Crispim Nascimento, Rossi, Erik Aranha, Paredes, Bruno Diaz, Bastos, Tanira Matutino, Meira, Cássio Santana, Nonaka, Carolina Vasques Kymie, Silva, Daniela Nascimento, Improta-Caria, Alex, Moreira, Diogo Rodrigo Magalhaes, Leite, Ana Cristina Lima, de Oliveira Filho, Gevanio Bezerra, Filho, José Maria Barbosa, dos Santos, Ricardo Ribeiro, Soares, Milena Botelho Pereira, Souza, Bruno Solano de Freita
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Hindawi 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8380504/
https://www.ncbi.nlm.nih.gov/pubmed/34434238
http://dx.doi.org/10.1155/2021/2642807
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author Portella, Diogo Crispim Nascimento
Rossi, Erik Aranha
Paredes, Bruno Diaz
Bastos, Tanira Matutino
Meira, Cássio Santana
Nonaka, Carolina Vasques Kymie
Silva, Daniela Nascimento
Improta-Caria, Alex
Moreira, Diogo Rodrigo Magalhaes
Leite, Ana Cristina Lima
de Oliveira Filho, Gevanio Bezerra
Filho, José Maria Barbosa
dos Santos, Ricardo Ribeiro
Soares, Milena Botelho Pereira
Souza, Bruno Solano de Freita
author_facet Portella, Diogo Crispim Nascimento
Rossi, Erik Aranha
Paredes, Bruno Diaz
Bastos, Tanira Matutino
Meira, Cássio Santana
Nonaka, Carolina Vasques Kymie
Silva, Daniela Nascimento
Improta-Caria, Alex
Moreira, Diogo Rodrigo Magalhaes
Leite, Ana Cristina Lima
de Oliveira Filho, Gevanio Bezerra
Filho, José Maria Barbosa
dos Santos, Ricardo Ribeiro
Soares, Milena Botelho Pereira
Souza, Bruno Solano de Freita
author_sort Portella, Diogo Crispim Nascimento
collection PubMed
description Chagas disease is caused by Trypanosoma cruzi infection and remains a relevant cause of chronic heart failure in Latin America. The pharmacological arsenal for Chagas disease is limited, and the available anti-T. cruzi drugs are not effective when administered during the chronic phase. Cardiomyocytes derived from human-induced pluripotent stem cells (hiPSC-CMs) have the potential to accelerate the process of drug discovery for Chagas disease, through predictive preclinical assays in target human cells. Here, we aimed to establish a novel high-content screening- (HCS-) based method using hiPSC-CMs to simultaneously evaluate anti-T. cruzi activity and cardiotoxicity of chemical compounds. To provide proof-of-concept data, the reference drug benznidazole and three compounds with known anti-T. cruzi activity (a betulinic acid derivative named BA5 and two thiazolidinone compounds named GT5A and GT5B) were evaluated in the assay. hiPSC-CMs were infected with T. cruzi and incubated for 48 h with serial dilutions of the compounds for determination of EC50 and CC50 values. Automated multiparametric analyses were performed using an automated high-content imaging system. Sublethal toxicity measurements were evaluated through morphological measurements related to the integrity of the cytoskeleton by phalloidin staining, nuclear score by Hoechst 33342 staining, mitochondria score following MitoTracker staining, and quantification of NT-pro-BNP, a peptide released upon mechanical myocardial stress. The compounds showed EC(50) values for anti-T. cruzi activity similar to those previously described for other cell types, and GT5B showed a pronounced trypanocidal activity in hiPSC-CMs. Sublethal changes in cytoskeletal and nucleus scores correlated with NT-pro-BNP levels in the culture supernatant. Mitochondrial score changes were associated with increased cytotoxicity. The assay was feasible and allowed rapid assessment of anti-T. cruzi action of the compounds, in addition to cardiotoxicity parameters. The utilization of hiPSC-CMs in the drug development workflow for Chagas disease may help in the identification of novel compounds.
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spelling pubmed-83805042021-08-24 A Novel High-Content Screening-Based Method for Anti-Trypanosoma cruzi Drug Discovery Using Human-Induced Pluripotent Stem Cell-Derived Cardiomyocytes Portella, Diogo Crispim Nascimento Rossi, Erik Aranha Paredes, Bruno Diaz Bastos, Tanira Matutino Meira, Cássio Santana Nonaka, Carolina Vasques Kymie Silva, Daniela Nascimento Improta-Caria, Alex Moreira, Diogo Rodrigo Magalhaes Leite, Ana Cristina Lima de Oliveira Filho, Gevanio Bezerra Filho, José Maria Barbosa dos Santos, Ricardo Ribeiro Soares, Milena Botelho Pereira Souza, Bruno Solano de Freita Stem Cells Int Research Article Chagas disease is caused by Trypanosoma cruzi infection and remains a relevant cause of chronic heart failure in Latin America. The pharmacological arsenal for Chagas disease is limited, and the available anti-T. cruzi drugs are not effective when administered during the chronic phase. Cardiomyocytes derived from human-induced pluripotent stem cells (hiPSC-CMs) have the potential to accelerate the process of drug discovery for Chagas disease, through predictive preclinical assays in target human cells. Here, we aimed to establish a novel high-content screening- (HCS-) based method using hiPSC-CMs to simultaneously evaluate anti-T. cruzi activity and cardiotoxicity of chemical compounds. To provide proof-of-concept data, the reference drug benznidazole and three compounds with known anti-T. cruzi activity (a betulinic acid derivative named BA5 and two thiazolidinone compounds named GT5A and GT5B) were evaluated in the assay. hiPSC-CMs were infected with T. cruzi and incubated for 48 h with serial dilutions of the compounds for determination of EC50 and CC50 values. Automated multiparametric analyses were performed using an automated high-content imaging system. Sublethal toxicity measurements were evaluated through morphological measurements related to the integrity of the cytoskeleton by phalloidin staining, nuclear score by Hoechst 33342 staining, mitochondria score following MitoTracker staining, and quantification of NT-pro-BNP, a peptide released upon mechanical myocardial stress. The compounds showed EC(50) values for anti-T. cruzi activity similar to those previously described for other cell types, and GT5B showed a pronounced trypanocidal activity in hiPSC-CMs. Sublethal changes in cytoskeletal and nucleus scores correlated with NT-pro-BNP levels in the culture supernatant. Mitochondrial score changes were associated with increased cytotoxicity. The assay was feasible and allowed rapid assessment of anti-T. cruzi action of the compounds, in addition to cardiotoxicity parameters. The utilization of hiPSC-CMs in the drug development workflow for Chagas disease may help in the identification of novel compounds. Hindawi 2021-08-11 /pmc/articles/PMC8380504/ /pubmed/34434238 http://dx.doi.org/10.1155/2021/2642807 Text en Copyright © 2021 Diogo Crispim Nascimento Portella et al. https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Portella, Diogo Crispim Nascimento
Rossi, Erik Aranha
Paredes, Bruno Diaz
Bastos, Tanira Matutino
Meira, Cássio Santana
Nonaka, Carolina Vasques Kymie
Silva, Daniela Nascimento
Improta-Caria, Alex
Moreira, Diogo Rodrigo Magalhaes
Leite, Ana Cristina Lima
de Oliveira Filho, Gevanio Bezerra
Filho, José Maria Barbosa
dos Santos, Ricardo Ribeiro
Soares, Milena Botelho Pereira
Souza, Bruno Solano de Freita
A Novel High-Content Screening-Based Method for Anti-Trypanosoma cruzi Drug Discovery Using Human-Induced Pluripotent Stem Cell-Derived Cardiomyocytes
title A Novel High-Content Screening-Based Method for Anti-Trypanosoma cruzi Drug Discovery Using Human-Induced Pluripotent Stem Cell-Derived Cardiomyocytes
title_full A Novel High-Content Screening-Based Method for Anti-Trypanosoma cruzi Drug Discovery Using Human-Induced Pluripotent Stem Cell-Derived Cardiomyocytes
title_fullStr A Novel High-Content Screening-Based Method for Anti-Trypanosoma cruzi Drug Discovery Using Human-Induced Pluripotent Stem Cell-Derived Cardiomyocytes
title_full_unstemmed A Novel High-Content Screening-Based Method for Anti-Trypanosoma cruzi Drug Discovery Using Human-Induced Pluripotent Stem Cell-Derived Cardiomyocytes
title_short A Novel High-Content Screening-Based Method for Anti-Trypanosoma cruzi Drug Discovery Using Human-Induced Pluripotent Stem Cell-Derived Cardiomyocytes
title_sort novel high-content screening-based method for anti-trypanosoma cruzi drug discovery using human-induced pluripotent stem cell-derived cardiomyocytes
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8380504/
https://www.ncbi.nlm.nih.gov/pubmed/34434238
http://dx.doi.org/10.1155/2021/2642807
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