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...
Main Authors: | , , , , , , , , , , , , , , |
---|---|
Format: | Article |
Language: | English |
Published: |
Hindawi Limited
2021-01-01
|
Series: | Stem Cells International |
Online Access: | http://dx.doi.org/10.1155/2021/2642807 |
id |
doaj-9fae47b067404108b9d3c40fddce80ac |
---|---|
record_format |
Article |
spelling |
doaj-9fae47b067404108b9d3c40fddce80ac2021-08-23T01:32:10ZengHindawi LimitedStem Cells International1687-96782021-01-01202110.1155/2021/2642807A Novel High-Content Screening-Based Method for Anti-Trypanosoma cruzi Drug Discovery Using Human-Induced Pluripotent Stem Cell-Derived CardiomyocytesDiogo Crispim Nascimento Portella0Erik Aranha Rossi1Bruno Diaz Paredes2Tanira Matutino Bastos3Cássio Santana Meira4Carolina Vasques Kymie Nonaka5Daniela Nascimento Silva6Alex Improta-Caria7Diogo Rodrigo Magalhaes Moreira8Ana Cristina Lima Leite9Gevanio Bezerra de Oliveira Filho10José Maria Barbosa Filho11Ricardo Ribeiro dos Santos12Milena Botelho Pereira Soares13Bruno Solano de Freita Souza14Gonçalo Moniz InstituteGonçalo Moniz InstituteD’Or Institute for Research and Education (IDOR)Gonçalo Moniz InstituteGonçalo Moniz InstituteD’Or Institute for Research and Education (IDOR)Gonçalo Moniz InstitutePost-Graduate Program in Medicine and HealthGonçalo Moniz InstituteDepartment of Pharmaceutical SciencesDepartment of Pharmaceutical SciencesFederal University of ParaibaGonçalo Moniz InstituteGonçalo Moniz InstituteGonçalo Moniz InstituteChagas 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 EC50 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.http://dx.doi.org/10.1155/2021/2642807 |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Diogo Crispim Nascimento Portella Erik Aranha Rossi Bruno Diaz Paredes Tanira Matutino Bastos Cássio Santana Meira Carolina Vasques Kymie Nonaka Daniela Nascimento Silva Alex Improta-Caria Diogo Rodrigo Magalhaes Moreira Ana Cristina Lima Leite Gevanio Bezerra de Oliveira Filho José Maria Barbosa Filho Ricardo Ribeiro dos Santos Milena Botelho Pereira Soares Bruno Solano de Freita Souza |
spellingShingle |
Diogo Crispim Nascimento Portella Erik Aranha Rossi Bruno Diaz Paredes Tanira Matutino Bastos Cássio Santana Meira Carolina Vasques Kymie Nonaka Daniela Nascimento Silva Alex Improta-Caria Diogo Rodrigo Magalhaes Moreira Ana Cristina Lima Leite Gevanio Bezerra de Oliveira Filho José Maria Barbosa Filho Ricardo Ribeiro dos Santos Milena Botelho Pereira Soares Bruno Solano de Freita Souza A Novel High-Content Screening-Based Method for Anti-Trypanosoma cruzi Drug Discovery Using Human-Induced Pluripotent Stem Cell-Derived Cardiomyocytes Stem Cells International |
author_facet |
Diogo Crispim Nascimento Portella Erik Aranha Rossi Bruno Diaz Paredes Tanira Matutino Bastos Cássio Santana Meira Carolina Vasques Kymie Nonaka Daniela Nascimento Silva Alex Improta-Caria Diogo Rodrigo Magalhaes Moreira Ana Cristina Lima Leite Gevanio Bezerra de Oliveira Filho José Maria Barbosa Filho Ricardo Ribeiro dos Santos Milena Botelho Pereira Soares Bruno Solano de Freita Souza |
author_sort |
Diogo Crispim Nascimento Portella |
title |
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_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_sort |
novel high-content screening-based method for anti-trypanosoma cruzi drug discovery using human-induced pluripotent stem cell-derived cardiomyocytes |
publisher |
Hindawi Limited |
series |
Stem Cells International |
issn |
1687-9678 |
publishDate |
2021-01-01 |
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 EC50 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. |
url |
http://dx.doi.org/10.1155/2021/2642807 |
work_keys_str_mv |
AT diogocrispimnascimentoportella anovelhighcontentscreeningbasedmethodforantitrypanosomacruzidrugdiscoveryusinghumaninducedpluripotentstemcellderivedcardiomyocytes AT erikaranharossi anovelhighcontentscreeningbasedmethodforantitrypanosomacruzidrugdiscoveryusinghumaninducedpluripotentstemcellderivedcardiomyocytes AT brunodiazparedes anovelhighcontentscreeningbasedmethodforantitrypanosomacruzidrugdiscoveryusinghumaninducedpluripotentstemcellderivedcardiomyocytes AT taniramatutinobastos anovelhighcontentscreeningbasedmethodforantitrypanosomacruzidrugdiscoveryusinghumaninducedpluripotentstemcellderivedcardiomyocytes AT cassiosantanameira anovelhighcontentscreeningbasedmethodforantitrypanosomacruzidrugdiscoveryusinghumaninducedpluripotentstemcellderivedcardiomyocytes AT carolinavasqueskymienonaka anovelhighcontentscreeningbasedmethodforantitrypanosomacruzidrugdiscoveryusinghumaninducedpluripotentstemcellderivedcardiomyocytes AT danielanascimentosilva anovelhighcontentscreeningbasedmethodforantitrypanosomacruzidrugdiscoveryusinghumaninducedpluripotentstemcellderivedcardiomyocytes AT aleximprotacaria anovelhighcontentscreeningbasedmethodforantitrypanosomacruzidrugdiscoveryusinghumaninducedpluripotentstemcellderivedcardiomyocytes AT diogorodrigomagalhaesmoreira anovelhighcontentscreeningbasedmethodforantitrypanosomacruzidrugdiscoveryusinghumaninducedpluripotentstemcellderivedcardiomyocytes AT anacristinalimaleite anovelhighcontentscreeningbasedmethodforantitrypanosomacruzidrugdiscoveryusinghumaninducedpluripotentstemcellderivedcardiomyocytes AT gevaniobezerradeoliveirafilho anovelhighcontentscreeningbasedmethodforantitrypanosomacruzidrugdiscoveryusinghumaninducedpluripotentstemcellderivedcardiomyocytes AT josemariabarbosafilho anovelhighcontentscreeningbasedmethodforantitrypanosomacruzidrugdiscoveryusinghumaninducedpluripotentstemcellderivedcardiomyocytes AT ricardoribeirodossantos anovelhighcontentscreeningbasedmethodforantitrypanosomacruzidrugdiscoveryusinghumaninducedpluripotentstemcellderivedcardiomyocytes AT milenabotelhopereirasoares anovelhighcontentscreeningbasedmethodforantitrypanosomacruzidrugdiscoveryusinghumaninducedpluripotentstemcellderivedcardiomyocytes AT brunosolanodefreitasouza anovelhighcontentscreeningbasedmethodforantitrypanosomacruzidrugdiscoveryusinghumaninducedpluripotentstemcellderivedcardiomyocytes AT diogocrispimnascimentoportella novelhighcontentscreeningbasedmethodforantitrypanosomacruzidrugdiscoveryusinghumaninducedpluripotentstemcellderivedcardiomyocytes AT erikaranharossi novelhighcontentscreeningbasedmethodforantitrypanosomacruzidrugdiscoveryusinghumaninducedpluripotentstemcellderivedcardiomyocytes AT brunodiazparedes novelhighcontentscreeningbasedmethodforantitrypanosomacruzidrugdiscoveryusinghumaninducedpluripotentstemcellderivedcardiomyocytes AT taniramatutinobastos novelhighcontentscreeningbasedmethodforantitrypanosomacruzidrugdiscoveryusinghumaninducedpluripotentstemcellderivedcardiomyocytes AT cassiosantanameira novelhighcontentscreeningbasedmethodforantitrypanosomacruzidrugdiscoveryusinghumaninducedpluripotentstemcellderivedcardiomyocytes AT carolinavasqueskymienonaka novelhighcontentscreeningbasedmethodforantitrypanosomacruzidrugdiscoveryusinghumaninducedpluripotentstemcellderivedcardiomyocytes AT danielanascimentosilva novelhighcontentscreeningbasedmethodforantitrypanosomacruzidrugdiscoveryusinghumaninducedpluripotentstemcellderivedcardiomyocytes AT aleximprotacaria novelhighcontentscreeningbasedmethodforantitrypanosomacruzidrugdiscoveryusinghumaninducedpluripotentstemcellderivedcardiomyocytes AT diogorodrigomagalhaesmoreira novelhighcontentscreeningbasedmethodforantitrypanosomacruzidrugdiscoveryusinghumaninducedpluripotentstemcellderivedcardiomyocytes AT anacristinalimaleite novelhighcontentscreeningbasedmethodforantitrypanosomacruzidrugdiscoveryusinghumaninducedpluripotentstemcellderivedcardiomyocytes AT gevaniobezerradeoliveirafilho novelhighcontentscreeningbasedmethodforantitrypanosomacruzidrugdiscoveryusinghumaninducedpluripotentstemcellderivedcardiomyocytes AT josemariabarbosafilho novelhighcontentscreeningbasedmethodforantitrypanosomacruzidrugdiscoveryusinghumaninducedpluripotentstemcellderivedcardiomyocytes AT ricardoribeirodossantos novelhighcontentscreeningbasedmethodforantitrypanosomacruzidrugdiscoveryusinghumaninducedpluripotentstemcellderivedcardiomyocytes AT milenabotelhopereirasoares novelhighcontentscreeningbasedmethodforantitrypanosomacruzidrugdiscoveryusinghumaninducedpluripotentstemcellderivedcardiomyocytes AT brunosolanodefreitasouza novelhighcontentscreeningbasedmethodforantitrypanosomacruzidrugdiscoveryusinghumaninducedpluripotentstemcellderivedcardiomyocytes |
_version_ |
1721199041184792576 |