Publicação
Development of novel antibody-drug conjugated molecules for treatment of B-cell maligancies
| Resumo: | Cancer is one of the leading causes of death worldwide, and is expected to continue increasing, with a projected 28.4 million cases by 2040. Dogs are also significantly affected by cancer, including NHL, which is one of the most common hematological malignancies in both species. Dogs are considered excellent models to accelerate the translation of treatments for human patients due to their similarities with humans. While immunotherapies, particularly monoclonal antibodies (mAbs), have brought specificity to cancer therapies, they are still mostly used in combination with conventional chemotherapy, which remains the standard of care for both species. However, conventional treatments still do not fully cure and have numerous adverse effects, highlighting the need for further improvement. To address this need, using the promising advantages of rabbit-derived sdAbs, we aimed to develop a platform for a novel sdAb drug delivery system for NHL treatment using the dog as an animal model. For this purpose, we explored three different drug delivery methods: ADCs, immunotoxins and immunoliposomes. For the development of ADC, we explored the potential of rabbit derived single-domain antibodies (sdAb) to selectively conjugate a payload towards cysteine at position 80. First, a rabbit-derived sdAb library against canine B-cell lymphoma receptors was subjected to in vitro and in vivo phage display. Then, VL sdAb that specifically targeted canine lymphoma cells in vitro and presented a good tumor uptake was selected for SN-38 site selective payload conjugation via its Cys80 and generated a stable and homogeneous C5- DAB-SN-38. This study validated a platform to develop novel ADCs that combine rabbit sdAbs benefits with the advantages of canine lymphoma model. Furthermore, this previously characterized and validated VL sdAb was also used to develop a new immunotoxin for the treatment of canine B-cell lymphoma. For that purpose, VL sdAb was conjugated with the PE38 toxin truncated form and tested in vitro in a canine B-cell lymphoma cells and in in vivo in a xenograft mouse model of canine lymphoma. This study validated immunotoxins as a potential treatment for canine lymphoma. Lastly, to validate a new liposome for canine lymphoma, we aimed to develop a liposome-based nanocarrier for panobinostat, using folate-targeted and non-targeted formulations. Both formulations were evaluated in canine lymphoma cells, validating liposomes as an effective treatment for canine lymphoma. In the future, our goal is to conjugate our VL sdAb to the liposome to obtain an immunoliposome for canine B-cell lymphoma. In conclusion, all the work developed contributed to the understanding of the importance of using the dog as a model and how these animals can contribute for clinical translation in the immune-oncology field |
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| Autores principais: | André, Ana Filipa Santos |
| Assunto: | Single-domain antibodies Lymphoma Antibody-drug conjugates Immunotoxins Immunoliposomes Anticorpos de pequeno domínio Linfoma Anticorpos conjugados a fármacos Imunotoxinas Imunolipossomas |
| Ano: | 2023 |
| País: | Portugal |
| Tipo de documento: | tese de doutoramento |
| Tipo de acesso: | acesso aberto |
| Instituição associada: | Universidade de Lisboa |
| Idioma: | inglês |
| Origem: | Repositório da Universidade de Lisboa |
| Resumo: | Cancer is one of the leading causes of death worldwide, and is expected to continue increasing, with a projected 28.4 million cases by 2040. Dogs are also significantly affected by cancer, including NHL, which is one of the most common hematological malignancies in both species. Dogs are considered excellent models to accelerate the translation of treatments for human patients due to their similarities with humans. While immunotherapies, particularly monoclonal antibodies (mAbs), have brought specificity to cancer therapies, they are still mostly used in combination with conventional chemotherapy, which remains the standard of care for both species. However, conventional treatments still do not fully cure and have numerous adverse effects, highlighting the need for further improvement. To address this need, using the promising advantages of rabbit-derived sdAbs, we aimed to develop a platform for a novel sdAb drug delivery system for NHL treatment using the dog as an animal model. For this purpose, we explored three different drug delivery methods: ADCs, immunotoxins and immunoliposomes. For the development of ADC, we explored the potential of rabbit derived single-domain antibodies (sdAb) to selectively conjugate a payload towards cysteine at position 80. First, a rabbit-derived sdAb library against canine B-cell lymphoma receptors was subjected to in vitro and in vivo phage display. Then, VL sdAb that specifically targeted canine lymphoma cells in vitro and presented a good tumor uptake was selected for SN-38 site selective payload conjugation via its Cys80 and generated a stable and homogeneous C5- DAB-SN-38. This study validated a platform to develop novel ADCs that combine rabbit sdAbs benefits with the advantages of canine lymphoma model. Furthermore, this previously characterized and validated VL sdAb was also used to develop a new immunotoxin for the treatment of canine B-cell lymphoma. For that purpose, VL sdAb was conjugated with the PE38 toxin truncated form and tested in vitro in a canine B-cell lymphoma cells and in in vivo in a xenograft mouse model of canine lymphoma. This study validated immunotoxins as a potential treatment for canine lymphoma. Lastly, to validate a new liposome for canine lymphoma, we aimed to develop a liposome-based nanocarrier for panobinostat, using folate-targeted and non-targeted formulations. Both formulations were evaluated in canine lymphoma cells, validating liposomes as an effective treatment for canine lymphoma. In the future, our goal is to conjugate our VL sdAb to the liposome to obtain an immunoliposome for canine B-cell lymphoma. In conclusion, all the work developed contributed to the understanding of the importance of using the dog as a model and how these animals can contribute for clinical translation in the immune-oncology field |
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