A via dos esfingolipídeos como biossensor da quimiossensibilidade do câncer: Uma prova de princípio

Cancer is a complex genetic disease with reduced treatment alternatives due to tumor heterogeneity and drug multiresistance emergence. The sphingolipid (SL) metabolic pathway integrates different responses of cellular stress signals and defines cell survival. Therefore, we suggest studying the perturbations on the sphingolipid pathway (SLP) caused by chemotherapeutic drugs using a systems biology approach to evaluate its functionality as a drug response sensor. We used a sphingomyelin-BODIPY (SM-BOD) sensor to study SL metabolism by flow cytometry and live cell imaging in different cancer models. To decode pathway complexity, we implemented Gussian mixture models, ordinary differential equations models, unsupervised classification algorithms and a fuzzy logic approach to assess its utility as a chemotherapy response sensor. Our results show that chemotherapeutic drugs perturb the SLP in different ways in a cell line-specific manner. In addition, we found that few SM-BOD fluorescence features predict chemosensitivity with high accuracy. Finally, we found that the relative species composition of SL appears to contribute to the resulting cytotoxicity of many treatments. This report offers a conceptual and mathematical framework for developing personalized mathematical models to predict and improve cancer therapy.El cáncer es una enfermedad genética compleja con opciones terapéuticas limitadas, debido a la heterogeneidad tumoral y a la aparición de multirresistencia a los fármacos. La vía metabólica de los esfingolípidos (SL) se caracteriza por ser capaz de integrar diferentes tipos de señales de estrés celular y definir la supervivencia celular. Por lo tanto, sugerimos estudiar las perturbaciones en la vía de los esfingolípidos (SLP) causadas por fármacos quimioterapéuticos utilizando un enfoque de biología de sistemas y evaluar su funcionalidad como sensor de respuesta a fármacos. Usamos un sensor de esfingomielina-BODIPY (SM-BOD) para estudiar el metabolismo de SL mediante citometría de flujo e imágenes de células vivas en diferentes modelos de cáncer. Para decodificar la complejidad de la ruta, implementamos modelos de mezcla gaussianos, modelos de ecuaciones diferenciales ordinarias, algoritmos de clasificación no supervisados ​y un modelo de lógica difusa para evaluar su utilidad como sensor de respuesta a la quimioterapia. Nuestros resultados muestran que los fármacos quimioterapéuticos perturban la SLP de diferentes formas y de una manera específica de la línea celular. Además, encontramos que pocas características de fluorescencia de SM-BOD predicen la quimiosensibilidad con alta precisión. Finalmente, encontramos que la composición relativa de especies de SL parece contribuir a la citotoxicidad resultante de muchos tratamientos. Este informe ofrece un marco conceptual y matemático para desarrollar modelos matemáticos personalizados para predecir y mejorar la terapia del cáncer.O câncer é uma doença genética complexa com opções terapêuticas limitadas, devido à heterogeneidade tumoral e ao aparecimento de multirresistência aos fármacos. A via metabólica dos esfingolipídeos (SP) é caracterizada por ser capaz de integrar diferentes tipos de sinais de estresse celular e definir a sobrevivência celular. Portanto, sugerimos estudar as perturbações na rota dos esfingolipídeos (SP) causadas por fármacos quimioterápicos usando uma abordagem de biologia de sistemas e avaliar sua funcionalidade como sensor de resposta a fármacos. Usamos um sensor de esfingomielina-BODIPY (SM-BOD) para estudar o metabolismo do SP por citometria de fluxo e imagens de células vivas em diferentes modelos de câncer. Para decodificar a complexidade do caminho, implementamos modelos de mistura gaussiana, modelos de equações diferenciais ordinárias, algoritmos de classificação não supervisionados ​e um modelo de lógica difusa para avaliar sua utilidade como sensor de resposta à quimioterapia. Nossos resultados mostram que os fármacos quimioterápicos perturbam a rota dos SP de diferentes maneiras e de maneira específica da linhagem celular. Além disso, descobrimos que poucas características de fluorescência do SM-BOD predizem a quimiossensibilidade com alta precisão. Finalmente, descobrimos que a composição relativa de espécies de SP parece contribuir para a citotoxicidade resultante de muitos tratamentos. Este relatório oferece uma estrutura conceitual e matemática para o desenvolvimento de modelos matemáticos personalizados para prever e melhorar a terapia do câncer

Perturbation-Based Modeling Unveils the Autophagic Modulation of Chemosensitivity and Immunogenicity in Breast Cancer Cells

In the absence of new therapeutic strategies, chemotherapeutic drugs are the most widely used strategy against metastatic breast cancer, in spite of eliciting multiple adverse effects and having low responses with an average 5-year patient survival rate. Among the new therapeutic targets that are currently in clinical trials, here, we addressed the association between the regulation of the metabolic process of autophagy and the exposure of damage-associated molecular patterns associated (DAMPs) to immunogenic cell death (ICD), which has not been previously studied. After validating an mCHR-GFP tandem LC3 sensor capacity to report dynamic changes of the autophagic metabolic flux in response to external stimuli and demonstrating that both basal autophagy levels and response to diverse autophagy regulators fluctuate among different cell lines, we explored the interaction between autophagy modulators and chemotherapeutic agents in regards of cytotoxicity and ICD using three different breast cancer cell lines. Since these interactions are very complex and variable throughout different cell lines, we designed a perturbation-based model in which we propose specific modes of action of chemotherapeutic agents on the autophagic flux and the corresponding strategies of modulation to enhance the response to chemotherapy. Our results point towards a promising therapeutic potential of the metabolic regulation of autophagy to overcome chemotherapy resistance by eliciting ICD.Universidad de Costa Rica/[803-B6-600]/UCR/Costa RicaUCR::Vicerrectoría de Investigación::Unidades de Investigación::Ciencias de la Salud::Centro de Investigación en Enfermedades Tropicales (CIET)UCR::Vicerrectoría de Docencia::Salud::Facultad de Microbiologí

Perturbation-Based Modeling Unveils the Autophagic Modulation of Chemosensitivity and Immunogenicity in Breast Cancer Cells

In the absence of new therapeutic strategies, chemotherapeutic drugs are the most widely used strategy against metastatic breast cancer, in spite of eliciting multiple adverse effects and having low responses with an average 5-year patient survival rate. Among the new therapeutic targets that are currently in clinical trials, here, we addressed the association between the regulation of the metabolic process of autophagy and the exposure of damage-associated molecular patterns associated (DAMPs) to immunogenic cell death (ICD), which has not been previously studied. After validating an mCHR-GFP tandem LC3 sensor capacity to report dynamic changes of the autophagic metabolic flux in response to external stimuli and demonstrating that both basal autophagy levels and response to diverse autophagy regulators fluctuate among different cell lines, we explored the interaction between autophagy modulators and chemotherapeutic agents in regards of cytotoxicity and ICD using three different breast cancer cell lines. Since these interactions are very complex and variable throughout different cell lines, we designed a perturbation-based model in which we propose specific modes of action of chemotherapeutic agents on the autophagic flux and the corresponding strategies of modulation to enhance the response to chemotherapy. Our results point towards a promising therapeutic potential of the metabolic regulation of autophagy to overcome chemotherapy resistance by eliciting ICD