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

Metabolites ◽  
2021 ◽  
Vol 11 (9) ◽  
pp. 637
Author(s):  
Isaac Quiros-Fernandez ◽  
Lucía Figueroa-Protti ◽  
Jorge L. Arias-Arias ◽  
Norman Brenes-Cordero ◽  
Francisco Siles ◽  
...  

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.

2019 ◽  
Vol 3 (Supplement_1) ◽  
Author(s):  
Violet Kiesel ◽  
Stephen Hursting ◽  
Dorothy Teegarden

Abstract Objectives Prevention of metastasis is of utmost importance for increasing survival in breast cancer patients. Oxygen tension is variable throughout tumors, creating regions of hypoxia that have been linked with poor cancer prognosis. Hypoxia increases glycolytic flux via hypoxia-inducible factor-1α (HIF1α), and can therefore alter growth and survival of cancer cells. Our objectives are to (1) characterize changes in metabolism and survival that occur when metastatic and non-metastatic mammary cancer cell lines are cultured in hypoxia, and (2) determine whether 1,25-dihydroxyvitamin D (1,25(OH)2D) reduces overall survival in hypoxia. Methods We utilized Wnt oncogene-driven murine mammary cancer cells that are non-metastatic (M-Wnt) or that preferentially metastasize to the lung in vivo (metM-Wntlung). Viability of M-Wnt and metM-Wntlung cells treated with 10 nM 1,25(OH)2D and/or 20 mM 2-deoxyglucose (2DG, an inhibitor of glycolysis) was measured with MTT. Expression of HIF1α protein was determined by Western blotting. Results We show that 1,25(OH)2D treatment significantly decreased viability of metastatic metM-Wntlung cells grown in hypoxia by 41%, whereas viability of M-Wnt cells was not significantly impacted by 1,25(OH)2D treatment. Furthermore, treating cells with 2DG significantly decreased viability of both cells lines in hypoxia, with metM-Wntlung cells being more sensitive to 2DG. Interestingly, 1,25(OH)2D treatment partially rescued M-Wnt cells by 22% and metM-Wntlung cells by 24% when treated with 2DG in hypoxia. Finally, we show that M-Wnt cells have 1.9-fold increased expression of HIF1α protein compared to metM-Wntlung cells when grown in hypoxia. Conclusions Our results collectively suggest that non-metastatic M-Wnt cells are less sensitive to treatment with 1,25(OH)2D and 2DG in hypoxia than metastatic metM-Wntlungcells. These data may be explained, in part, by elevated expression of HIF1α in M-Wnt cells, which may contribute to their improved survival in hypoxia. Funding Sources National Institute of Health and USDA.


Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 2557-2557
Author(s):  
Oxana Norkina ◽  
Archana Thakur ◽  
Maxim Norkin ◽  
Elyse Paul ◽  
Zaid Al-Kadhimi ◽  
...  

Abstract In our ongoing phase I dose-escalation trial in women with metastatic breast cancer, multiple infusions of ATC armed with Her2Bi (aATC) induced elevated levels of IL-2, IFNγ TNFα, GM-CSF, and IL-12 as well as the development of cytotoxic activity directed at breast cancer cell lines lasting up to 4 months (Grabert et al. Clin Cancer Res. 2006). These results suggested that aATC infusions “vaccinated” the endogenous immune system of the patients. In this study, we explored mechanisms of the increased cytotoxicity after immunotherapy with armed ATC by testing for cell-mediated cytotoxicity and in vitro anti-tumor antibody synthesis. We co-cultured irradiated aATC with fresh PBMC in presence or absence of SKBR3 for 7 days, collected the PBMC, and tested for cytotoxicity in MTT and Cr51 release assays directed at SKBR3, Daudi and A-431 cell lines. PBMC mediated high levels of non-specific cytotoxicity against all tested cell lines and there were no phenotypic changes in the PBMC after 7 days of co-culture. When PBMC were co-cultured with irradiated aATC and SKBR3 for 21 days in presence of IL-2, the B cells in the PBMC produced significantly higher amounts of specific Abs directed SKBR3 (ELISA for antibodies binding to SKBR3) compared to PBMC co-cultured with SKBR3 alone. CpG ODN type C augmented in vitro anti-SKBR3 Ab synthesis. These studies show that Her2Bi-armed ATCco-cultured with PBMC enhanced nonspecific cytotoxicity and induced in vitro specific antibody synthesis directed at SKBR3 cells. Evidence that Her2Bi armed ATC can induce a vaccination response is supported by dendritic cell (DC) loading experiments in which aATC were transiently mixed with SKBR3 cells to generate tumor lysate. IL-4 and GM-CSF generated DC were exposed to the lysate for 24 h, washed, and co-cultured with fresh PBMC for 14 and 21 days. At the end of co-culture, cytotoxicity assays against SKBR3 increased markedly whereas cytotoxicity directed at Daudi targets was low. In addition, there were considerable levels of Abs directed to SKBR3 in the supernatants of PBMC co-cultured with DC loaded with SKBR3 lysate, but not with DC loaded with SKBR3-culture media or RPMI alone. These studies established that the lysate produced as a result of aATC cytotoxicity against SKBR3 is immunogenic for DC. In summary, when PBMC were co-cultured with DC exposed to SKBR3 lysate, there was induction of specific cytotoxicity and in vitro tumor specific Abs synthesis. Together with experiments involving primary co-cultures of irradiated aATC, PBMC, and SKBR3, our studies show that there are both non-specific and specific cellular and humoral responses generated as a result of co-culture with Her2Bi armed ATC. These studies provide evidence that aATC infusions can induce both specific and non-specific cellular, humoral, and cytokine responses from the endogenous immune systems of patients. Please credit the grants R01CA92344, 5P30CA022453-25, 1819 from Michigan Economic Development Corporation and 6066-06 from Leukemia and Lymphoma Society


2002 ◽  
Vol 96 (Sup 2) ◽  
pp. A75
Author(s):  
Michiko Yamaguchi ◽  
Toshiya Tsujita ◽  
Hiroshi Miyoshi ◽  
Sachiko Todoroki ◽  
Koji Sumikawa

2020 ◽  
Author(s):  
Joy Ifunanya Odimegwu ◽  
Olukemi Abiodun Odukoya ◽  
Alejandro Español ◽  
Maria Elena Sales

ABSTRACTObjectiveWe aim to test the efficacy of edible Dioscorea species grown and consumed in Nigeria, Africa on two breast cancer cell lines; MCF-7 and MDA-MB231 derived from a luminal and a triple-negative breast cancer (TNBC) respectively and to confirm safety in non-tumour cells MCF-10A using a well established cytotoxic compound paclitaxel as a standard. Metastatic breast cancer is a prevalent cause of mortality in women around the world. Breast cancer therapies have greatly advanced in recent years, but many patients develop cancer re-occurrence and metastasis and subsequently yield to the disease because of chemoresistance.MethodsEthanolic extracts of Dioscorea rotundata boiled and raw (DiosB and DiosR) respectively were chemically analysed for the presence of diosgenin using HPLC and the cytotoxic activity of the extracts were tested on MCF-7, MDA-MB-231 and MCF-10A cells In vitro by MTT assay.ResultsDiosB and DiosR extracts showed a higher maximal effect on MCF-7 cells than on MDA-MB231 after 24 h and 48 h treatments (p<0.0001 and p<0.05 respectively). DiosR, if applied at a range between 50-70 g/ml, can be effective to reduce breast tumor cell viability without affecting non tumorigenic MCF-10A cells either at 24 h or at 48 h. DiosB showed an IC50 of 38.83μg/ml while DiosR showed an IC50 of 41.80μg/ml.ConclusionThese results show that ethanolic extracts of Dioscorea rotundata tubers could be used effectively to treat breast cancer tumors and this is in sync with its diosgenin content as other Dioscorea species applied for similar treatments in Asia and elsewhere.


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