Publicação
Development of dual-targeting immunoliposomes against canine B cell lymphoma
| Resumo: | Non-Hodgkin lymphoma (NHL) is responsible for numerous cancer-related deaths worldwide, both in human and in veterinary medicine. Even though treatment options have improved, refractory/relapsed disease remains a clinical challenge, urging the development of safer and more efficient novel treatment strategies. Knowing that canine lymphoma (cNHL) shares many similarities with human NHL, both species would benefit from the use of dogs with spontaneous lymphoma as animal models for cancer research. In this context, in a recent study, we allied the use of liposomes, nanocarriers with great promise in drug delivery that increase selectivity and thus reduce adverse effects, to a powerful cytotoxic molecule, panobinostat, for the treatment of cNHL. In the present study, with the intention of enhancing this therapy’s selectivity, the liposomes were targeted through dual targeting with folic acid (FA) and a highly specific single domain antibody (sdAb) against cNHL, named C5. The single targeted (FA and C5) and nontargeted liposome formulations where simultaneously tested as reference. After optimization of the development of liposomes targeted with sdAb C5, the liposome formulations (Lip, Lip-Fa, Lip-C5, Lip-FA-C5) were evaluated for their targeting properties against a canine lymphoma cell line (CLBL-1). The preliminary results showed that dual targeting did not enhance the cell internalization in comparison with single targeting, nonetheless all targeted formulations presented a higher targeting compared with nontargeted liposomes. Subsequently, the cytotoxic activity of the encapsulated panobinostat was evaluated in vitro. Results showed a half maximal inhibitory concentration (IC50) in the low nanomolar range: 20,8 nM for dual-targeted liposomes and 30,4 nM for the single targeted C5 formulation. Additionally, this study confirmed that the encapsulated panobinostat’s cytotoxicity mechanism is associated with histone H3 acetylation. To evaluate the targeting properties in vivo of the studied liposomes, a preliminary in vivo biodistribution assay using indium-111 was performed for untargeted and folate targeted panobinostat liposomes. Targeted liposomes showed a tumor uptake two-fold higher than the nontargeted formulation (2,5% and 0,91% ID/g respectively), confirming the promise of this safer and more effective treatment approach. This study demonstrated the targeting and activity of dual-targeted Panobinostat liposomes against CL, confirming this strategy’s promise |
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| Autores principais: | Leonardo, Ana Sofia Rodrigues |
| Assunto: | Canine lymphoma Liposomes Single-domain antibodies Folic acid Panobinostat Linfoma canino Lipossomas Anticorpos de domínio único Ácido fólico Panobinostat |
| Ano: | 2022 |
| 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: | Non-Hodgkin lymphoma (NHL) is responsible for numerous cancer-related deaths worldwide, both in human and in veterinary medicine. Even though treatment options have improved, refractory/relapsed disease remains a clinical challenge, urging the development of safer and more efficient novel treatment strategies. Knowing that canine lymphoma (cNHL) shares many similarities with human NHL, both species would benefit from the use of dogs with spontaneous lymphoma as animal models for cancer research. In this context, in a recent study, we allied the use of liposomes, nanocarriers with great promise in drug delivery that increase selectivity and thus reduce adverse effects, to a powerful cytotoxic molecule, panobinostat, for the treatment of cNHL. In the present study, with the intention of enhancing this therapy’s selectivity, the liposomes were targeted through dual targeting with folic acid (FA) and a highly specific single domain antibody (sdAb) against cNHL, named C5. The single targeted (FA and C5) and nontargeted liposome formulations where simultaneously tested as reference. After optimization of the development of liposomes targeted with sdAb C5, the liposome formulations (Lip, Lip-Fa, Lip-C5, Lip-FA-C5) were evaluated for their targeting properties against a canine lymphoma cell line (CLBL-1). The preliminary results showed that dual targeting did not enhance the cell internalization in comparison with single targeting, nonetheless all targeted formulations presented a higher targeting compared with nontargeted liposomes. Subsequently, the cytotoxic activity of the encapsulated panobinostat was evaluated in vitro. Results showed a half maximal inhibitory concentration (IC50) in the low nanomolar range: 20,8 nM for dual-targeted liposomes and 30,4 nM for the single targeted C5 formulation. Additionally, this study confirmed that the encapsulated panobinostat’s cytotoxicity mechanism is associated with histone H3 acetylation. To evaluate the targeting properties in vivo of the studied liposomes, a preliminary in vivo biodistribution assay using indium-111 was performed for untargeted and folate targeted panobinostat liposomes. Targeted liposomes showed a tumor uptake two-fold higher than the nontargeted formulation (2,5% and 0,91% ID/g respectively), confirming the promise of this safer and more effective treatment approach. This study demonstrated the targeting and activity of dual-targeted Panobinostat liposomes against CL, confirming this strategy’s promise |
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