Publicação
Synthesis, characterization, and biological evaluation of peptides capable of interfering with RANK-TRAF6 pathway using nuclear imaging
| Resumo: | The RANK-RANKL interaction, and consequently, the RANK-TRAF6 pathway, is involved in several fundamental biological cycles, such as cell growth, apoptosis, osteosteoclastogenesis and inflammatory responses. As a result of partaking in such important processes, it is also implied that this pathway is related to severe pathologies, such as osteoporosis and breast or prostate cancer-induced bone metastasis. Currently, one of the therapeutic approaches to treat such diseases lies in the inhibition of RANK-RANKL interaction. Nonetheless, considerable interest arose in an alternative route, namely the inhibition of the RANK-TRAF6 pathway. This process is achievable, for example, by the interference of RANK’s interaction with TRAF6 using decoy peptides (e.g., L-T6DP-1). 8 Thus, the objective of this project was to synthesize, purify and biologically evaluate two peptides (3A and its negative control, 3B) containing a cell penetrating sequence (KLFMALVAFLRFLT), combined with sequences designed in silico to interfere with the RANK-TRAF6 pathway (RQMATADEA and RQMPTEDEY). Furthermore, biodistribution studies of peptides 3A and 3B, modified with a gallium-67 chelator were aimed to be achieved, in in vivo animal models, by micro-PET-SPECT-CT imaging. The aforementioned studies were not performed, as during the process, several issues arose. Some include long amino acid coupling times and difficulty radiolabeling peptide 3A. These adversities led to an overall unsuccessful procedure, requiring further optimization. Although the image acquisition of the synthesized peptides was impossible at this time, it was deemed that the equipment validation should still be made using an alternative compound that is currently being investigated by the C2TN-IST group: a 67Ga-radiolabeled gold nanoparticle named 67Ga-AuNP-BBN-Pt1. |
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| Autores principais: | Gonçalves, Kyle Manuel Ferreira |
| Assunto: | Recetor ativador do fator nuclear kappa-B (RANK) Recetor-associado 6 do fator de necrose tumoral (TRAF6) Fator Nuclear-Kappa B (NF-κB) Síntese Peptídica em Fase Sólida (SPPS) Imagiologia Nuclear Teses de mestrado - 2023 |
| Ano: | 2023 |
| 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: | The RANK-RANKL interaction, and consequently, the RANK-TRAF6 pathway, is involved in several fundamental biological cycles, such as cell growth, apoptosis, osteosteoclastogenesis and inflammatory responses. As a result of partaking in such important processes, it is also implied that this pathway is related to severe pathologies, such as osteoporosis and breast or prostate cancer-induced bone metastasis. Currently, one of the therapeutic approaches to treat such diseases lies in the inhibition of RANK-RANKL interaction. Nonetheless, considerable interest arose in an alternative route, namely the inhibition of the RANK-TRAF6 pathway. This process is achievable, for example, by the interference of RANK’s interaction with TRAF6 using decoy peptides (e.g., L-T6DP-1). 8 Thus, the objective of this project was to synthesize, purify and biologically evaluate two peptides (3A and its negative control, 3B) containing a cell penetrating sequence (KLFMALVAFLRFLT), combined with sequences designed in silico to interfere with the RANK-TRAF6 pathway (RQMATADEA and RQMPTEDEY). Furthermore, biodistribution studies of peptides 3A and 3B, modified with a gallium-67 chelator were aimed to be achieved, in in vivo animal models, by micro-PET-SPECT-CT imaging. The aforementioned studies were not performed, as during the process, several issues arose. Some include long amino acid coupling times and difficulty radiolabeling peptide 3A. These adversities led to an overall unsuccessful procedure, requiring further optimization. Although the image acquisition of the synthesized peptides was impossible at this time, it was deemed that the equipment validation should still be made using an alternative compound that is currently being investigated by the C2TN-IST group: a 67Ga-radiolabeled gold nanoparticle named 67Ga-AuNP-BBN-Pt1. |
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