Publicação
Targeting necroptosis in human disease: potential novel modulators
| Resumo: | Necroptosis is an alternative pathway of regulated cell death, occurring independently of caspases, and mediated by receptor interacting serine/threonine-protein kinase-1 (RIP1), RIP3, and mixed lineage kinase domain-like (MLKL) protein. Necroptosis participates in the pathogenesis of several human conditions and activation may be beneficial for treating cancer. Despite this information, there are currently no pharmacological inhibitors or activators of necroptosis available with therapeutic properties. Colon cancer is the second most prevalent cancer worldwide and the third with the highest mortality rate. Cancer therapy has improved in recent years, but pro-apoptotic therapies still face cancer resistance. In this thesis, we started by screening three libraries of newly synthesized compounds for its ability to modulate necroptosis, using the murine fibrosarcoma L929 cell line as an in vitro model. We identified 7 compounds with the ability to inhibit TNF-α-induced necroptosis L929 cells by more than 70%. To further characterize compound activity, the half maximal effective concentration (EC50) for inhibiting necroptosis was determined. The compound with the lowest EC50 - MS-PSR90 - was further shown to reduce TNF-α-induced MLKL phosphorylation. Interestingly, one of the compounds - SAS9 - strongly induced necroptosis in both L929 cells and in human colorectal adenocarcinoma cells HT29; IC50 values were 16.96 and 49.73 μM, respectively. Interestingly, SAS9 concomitantly induced apoptosis in both cell lines, with increased caspase-3/7-like activity. In L929 cells, SAS9-induced necroptosis encompassed MLKL phosphorylation but not increased RIP1 expression. In conclusion, several novel modulators of necroptosis were identified in our screening, including a potential inducer of necroptosis. A better understanding of its mechanisms of action, as well as the investigation of pharmacokinetic properties, should help to determine their potential utility in cancer treatment. |
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| Autores principais: | Poim, Ana Rita Teixeira |
| Assunto: | Cancer therapy Drug screening Molecular targeting Necroptosis Teses de mestrado - 2017 |
| Ano: | 2017 |
| País: | Portugal |
| Tipo de documento: | dissertação de mestrado |
| Tipo de acesso: | acesso aberto |
| Instituição associada: | Universidade de Lisboa |
| Idioma: | inglês |
| Origem: | Repositório da Universidade de Lisboa |
| Resumo: | Necroptosis is an alternative pathway of regulated cell death, occurring independently of caspases, and mediated by receptor interacting serine/threonine-protein kinase-1 (RIP1), RIP3, and mixed lineage kinase domain-like (MLKL) protein. Necroptosis participates in the pathogenesis of several human conditions and activation may be beneficial for treating cancer. Despite this information, there are currently no pharmacological inhibitors or activators of necroptosis available with therapeutic properties. Colon cancer is the second most prevalent cancer worldwide and the third with the highest mortality rate. Cancer therapy has improved in recent years, but pro-apoptotic therapies still face cancer resistance. In this thesis, we started by screening three libraries of newly synthesized compounds for its ability to modulate necroptosis, using the murine fibrosarcoma L929 cell line as an in vitro model. We identified 7 compounds with the ability to inhibit TNF-α-induced necroptosis L929 cells by more than 70%. To further characterize compound activity, the half maximal effective concentration (EC50) for inhibiting necroptosis was determined. The compound with the lowest EC50 - MS-PSR90 - was further shown to reduce TNF-α-induced MLKL phosphorylation. Interestingly, one of the compounds - SAS9 - strongly induced necroptosis in both L929 cells and in human colorectal adenocarcinoma cells HT29; IC50 values were 16.96 and 49.73 μM, respectively. Interestingly, SAS9 concomitantly induced apoptosis in both cell lines, with increased caspase-3/7-like activity. In L929 cells, SAS9-induced necroptosis encompassed MLKL phosphorylation but not increased RIP1 expression. In conclusion, several novel modulators of necroptosis were identified in our screening, including a potential inducer of necroptosis. A better understanding of its mechanisms of action, as well as the investigation of pharmacokinetic properties, should help to determine their potential utility in cancer treatment. |
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