Miniheart: 3D culture of cardiac cells shows contractions, among other characteristics of the organ. Video: Seydel and collaborators, Biomedicines, 2024.
Bioengineering-based research developed at the Oswaldo Cruz Institute (IOC/Fiocruz) open up innovative possibilities in the search for treatments for cancer and Chagas disease.
The studies start with the creation of three-dimensional (3D) cell cultures, called spheroids or organoids, capable of reproducing characteristics of biological tissues.
Cancer and Chagas disease are the targets of research using this technology. Among the most recent results is the development of a model for testing personalized cancer therapies, called a tumor chemogram.
Work is also underway to identify the potential of a drug to treat cardiac fibrosis associated with infection by the parasite. trypanosoma cruzi, which causes Chagas disease.
“Expertise in tissue bioengineering is an advantage for studies on therapeutic response and pathophysiological mechanisms. Spheroids reproduce what occurs in the tissue in vivo in a way that is closer to reality than a two-dimensional cell culture in a monolayer,” explains the researcher at the Laboratory of Virology and Molecular Parasitology and current vice-director of Research, Technological Development and Innovation at the IOC, Luciana Garzoni, who is leading the studies.
The tumor chemogram development project will complete ten years in 2025. The initiative began in 2015, during doctoral research developed at Postgraduate Program in Cellular and Molecular Biology at IOC by the current postdoctoral student at the Laura Lacerda Coelho Institute, under the supervision of Luciana.
For the test, tumor organoids are produced in the laboratory from cells isolated from tumor fragments. Placed in culture dishes, these cells spontaneously reorganize into three-dimensional structures, forming microtumors.
As they reproduce the morphological and functional characteristics of tumors, they can be used to evaluate the power of action of different medications, identifying those with the best performance against the specific tumor of the patients.
In the image, a tumor spheroid presents a necrotic nucleus, as observed in cancer. Photo: Gutemberg Brito
According to the researcher, the objective is to enable personalized therapy, in search of a more effective treatment.
“The test is a breakthrough in the context of translational oncology, which seeks to apply advances in basic research to solve challenges in the diagnosis and treatment of cancer. Even if two patients have the same type of cancer, the way each tumor responds to therapy may vary. In this scenario, the purpose of the tumor chemogram is to evaluate which drug would be most effective for treating the patient using a sample of tumor cells from the patient himself,” he explains.
According to the researcher, the methodology for producing the tumor chemogram was tested in preclinical trials using cells collected directly from animal tumors, considered as experimental models. It was also validated with human cells from tumor lines, which are isolated from patients and adapted to grow continuously in the laboratory. These inputs are supplied by specialized companies and are widely used in research.
The innovation reached technological maturity level 6 (TRL 6), with demonstration of a fully functional prototype.
“To advance the technology, we need to begin large-scale testing under good laboratory practice (GLP) conditions, which will be done in the first half of 2025, using cells collected from patients’ tumors,” highlights the researcher.
The group recently published a study based on this methodology, which reproduced in the laboratory the process of breast tumor metastasis, which occurs when cancer spreads throughout the body.
Published in the scientific journal 'Biology', the study showed that the 3D model reproduces the migration of tumor cells and the change in expression of proteins essential for the initiation of the metastatic process, in a similar way to what occurs in tumors in patients.
“The epithelial cells of the spheroid acquire mesenchymal characteristics, becoming capable of migrating and detaching themselves from the tumor. This is one of the processes involved in the development of metastases, when tumor cells establish themselves in other tissues. Thus, we have an excellent model for evaluating new antimetastatic drugs,” says Luciana.
The work included tests with the drug doxorubicin, which is frequently used in the treatment of breast cancer and which has shown an important antimetastatic effect, inhibiting the epithelial-mesenchymal transition and the migration of tumor cells.
No IOC, the work was carried out in partnership with scientists from the laboratories of Innovations in Therapies, Teaching and Bioproducts; Research on the Thymus; Pathology; and Epidemiology of Congenital Malformations. Researchers from the Fluminense Federal University (UFF) and the National Cancer Institute (Inca) also participated.
According to the researcher, the next step is to advance in the clinical study of characterizing the tumor chemogram as a predictor of therapeutic response, in an environment that simulates real conditions of use, which is foreseen in the post-doctoral project developed by biologist Gabriela Vieira.
In addition to cancer research, studies on Chagas disease have advanced based on tests with organoids. In this type of study, one of the main advantages of the three-dimensional model is to reproduce the process of fibrosis and hypertrophy that affects the heart muscle in the chronic form of Chagas disease, as previously demonstrated in scientific articles published by the group.
In approximately 30% of patients, the presence of the parasite and the persistent inflammatory process in the heart damage the heart muscle, which leads to the development of fibrosis, with the production of scars. Excessive scarring causes changes in the architecture of the heart tissue, impairing its functioning and can cause problems such as arrhythmia and heart failure.
To better understand this process and seek new therapies, the researchers developed an organoid model called a mini-heart, formed by cardiac cells. With a spherical shape, cardiac organoids are 3D cell cultures capable of spontaneous contraction and cellular and molecular interactions similar to those observed in heart muscle.
“When we infect the mini hearts with T. cruzi, we were able to reproduce the fibrosis process not only at the molecular level, but also at the structural level. This allows us to test compounds to try to block or reverse this process”, highlights Luciana.
In experiments with minihearts infected by T. cruzi, the group has already identified compounds with antifibrotic activity. The most recent research was carried out by biomedical scientist Clara Seydel, under the guidance of Luciana, during her master's degree in the Postgraduate Program in Cellular and Molecular Biology at IOC. In the study, a positive result was observed with a medication, which managed to reduce hypertrophy and the expression of proteins involved in fibrosis in cardiac tissue.
“These results suggest that the model has great potential for studying new therapeutic approaches for treating cardiac fibrosis in Chagas disease,” comments Luciana. The researcher adds that the clinical use of the drugs already tested still depends on other stages of research, including testing on animals and patients.
Furthermore, mini-hearts can contribute to studies on the formation of cardiac blood vessels, since scientists have demonstrated that organoids produced with mesenchymal stem cells and endothelial cells develop blood vessels, indicating potential for studies in the field of translational cardiology.
Articles:
Coelho, LL; et al. Spheroid Model of Mammary Tumor Cells: Epithelial–Mesenchymal Transition and Doxorubicin Response. Biology 2024, 13, 463. https://doi.org/10.3390/biology13070463
Seydel, CM; et al. Exploring the Dimensions of Pre-Clinical Research: 3D Cultures as an Investigative Model of Cardiac Fibrosis in Chagas Disease. Biomedicines 2024, 12, 1410. https://doi.org/10.3390/biomedicines12071410
Cover image: Breast tumor spheroid in immunofluorescence microscopy image, extracted from the article by Coelho, LL; et al, published in the journal Biology, in 2024 (according to reference aima).
Miniheart: 3D culture of cardiac cells shows contractions, among other characteristics of the organ. Video: Seydel and collaborators, Biomedicines, 2024.
Bioengineering-based research developed at the Oswaldo Cruz Institute (IOC/Fiocruz) open up innovative possibilities in the search for treatments for cancer and Chagas disease.
The studies start with the creation of three-dimensional (3D) cell cultures, called spheroids or organoids, capable of reproducing characteristics of biological tissues.
Cancer and Chagas disease are the targets of research using this technology. Among the most recent results is the development of a model for testing personalized cancer therapies, called a tumor chemogram.
Work is also underway to identify the potential of a drug to treat cardiac fibrosis associated with infection by the parasite. trypanosoma cruzi, which causes Chagas disease.
“Expertise in tissue bioengineering is an advantage for studies on therapeutic response and pathophysiological mechanisms. Spheroids reproduce what occurs in the tissue in vivo in a way that is closer to reality than a two-dimensional cell culture in a monolayer,” explains the researcher at the Laboratory of Virology and Molecular Parasitology and current vice-director of Research, Technological Development and Innovation at the IOC, Luciana Garzoni, who is leading the studies.
The tumor chemogram development project will complete ten years in 2025. The initiative began in 2015, during doctoral research developed at Postgraduate Program in Cellular and Molecular Biology at IOC by the current postdoctoral student at the Laura Lacerda Coelho Institute, under the supervision of Luciana.
For the test, tumor organoids are produced in the laboratory from cells isolated from tumor fragments. Placed in culture dishes, these cells spontaneously reorganize into three-dimensional structures, forming microtumors.
As they reproduce the morphological and functional characteristics of tumors, they can be used to evaluate the power of action of different medications, identifying those with the best performance against the specific tumor of the patients.
In the image, a tumor spheroid presents a necrotic nucleus, as observed in cancer. Photo: Gutemberg Brito
According to the researcher, the objective is to enable personalized therapy, in search of a more effective treatment.
“The test is a breakthrough in the context of translational oncology, which seeks to apply advances in basic research to solve challenges in the diagnosis and treatment of cancer. Even if two patients have the same type of cancer, the way each tumor responds to therapy may vary. In this scenario, the purpose of the tumor chemogram is to evaluate which drug would be most effective for treating the patient using a sample of tumor cells from the patient himself,” he explains.
According to the researcher, the methodology for producing the tumor chemogram was tested in preclinical trials using cells collected directly from animal tumors, considered as experimental models. It was also validated with human cells from tumor lines, which are isolated from patients and adapted to grow continuously in the laboratory. These inputs are supplied by specialized companies and are widely used in research.
The innovation reached technological maturity level 6 (TRL 6), with demonstration of a fully functional prototype.
“To advance the technology, we need to begin large-scale testing under good laboratory practice (GLP) conditions, which will be done in the first half of 2025, using cells collected from patients’ tumors,” highlights the researcher.
The group recently published a study based on this methodology, which reproduced in the laboratory the process of breast tumor metastasis, which occurs when cancer spreads throughout the body.
Published in the scientific journal 'Biology', the study showed that the 3D model reproduces the migration of tumor cells and the change in expression of proteins essential for the initiation of the metastatic process, in a similar way to what occurs in tumors in patients.
“The epithelial cells of the spheroid acquire mesenchymal characteristics, becoming capable of migrating and detaching themselves from the tumor. This is one of the processes involved in the development of metastases, when tumor cells establish themselves in other tissues. Thus, we have an excellent model for evaluating new antimetastatic drugs,” says Luciana.
The work included tests with the drug doxorubicin, which is frequently used in the treatment of breast cancer and which has shown an important antimetastatic effect, inhibiting the epithelial-mesenchymal transition and the migration of tumor cells.
No IOC, the work was carried out in partnership with scientists from the laboratories of Innovations in Therapies, Teaching and Bioproducts; Research on the Thymus; Pathology; and Epidemiology of Congenital Malformations. Researchers from the Fluminense Federal University (UFF) and the National Cancer Institute (Inca) also participated.
According to the researcher, the next step is to advance in the clinical study of characterizing the tumor chemogram as a predictor of therapeutic response, in an environment that simulates real conditions of use, which is foreseen in the post-doctoral project developed by biologist Gabriela Vieira.
In addition to cancer research, studies on Chagas disease have advanced based on tests with organoids. In this type of study, one of the main advantages of the three-dimensional model is to reproduce the process of fibrosis and hypertrophy that affects the heart muscle in the chronic form of Chagas disease, as previously demonstrated in scientific articles published by the group.
In approximately 30% of patients, the presence of the parasite and the persistent inflammatory process in the heart damage the heart muscle, which leads to the development of fibrosis, with the production of scars. Excessive scarring causes changes in the architecture of the heart tissue, impairing its functioning and can cause problems such as arrhythmia and heart failure.
To better understand this process and seek new therapies, the researchers developed an organoid model called a mini-heart, formed by cardiac cells. With a spherical shape, cardiac organoids are 3D cell cultures capable of spontaneous contraction and cellular and molecular interactions similar to those observed in heart muscle.
“When we infect the mini hearts with T. cruzi, we were able to reproduce the fibrosis process not only at the molecular level, but also at the structural level. This allows us to test compounds to try to block or reverse this process”, highlights Luciana.
In experiments with minihearts infected by T. cruzi, the group has already identified compounds with antifibrotic activity. The most recent research was carried out by biomedical scientist Clara Seydel, under the guidance of Luciana, during her master's degree in the Postgraduate Program in Cellular and Molecular Biology at IOC. In the study, a positive result was observed with a medication, which managed to reduce hypertrophy and the expression of proteins involved in fibrosis in cardiac tissue.
“These results suggest that the model has great potential for studying new therapeutic approaches for treating cardiac fibrosis in Chagas disease,” comments Luciana. The researcher adds that the clinical use of the drugs already tested still depends on other stages of research, including testing on animals and patients.
Furthermore, mini-hearts can contribute to studies on the formation of cardiac blood vessels, since scientists have demonstrated that organoids produced with mesenchymal stem cells and endothelial cells develop blood vessels, indicating potential for studies in the field of translational cardiology.
Articles:
Coelho, LL; et al. Spheroid Model of Mammary Tumor Cells: Epithelial–Mesenchymal Transition and Doxorubicin Response. Biology 2024, 13, 463. https://doi.org/10.3390/biology13070463
Seydel, CM; et al. Exploring the Dimensions of Pre-Clinical Research: 3D Cultures as an Investigative Model of Cardiac Fibrosis in Chagas Disease. Biomedicines 2024, 12, 1410. https://doi.org/10.3390/biomedicines12071410
Cover image: Breast tumor spheroid in immunofluorescence microscopy image, extracted from the article by Coelho, LL; et al, published in the journal Biology, in 2024 (according to reference aima).
The non-profit reproduction of the text is allowed as long as the source is cited (Comunicação / Instituto Oswaldo Cruz)
 |
 |
