Publicação
Nanoheaters – Localized hyperthermia for precise gene delivery
| Resumo: | Gene therapy relies on the precise transfection of nucleic acid effectors into cancer cells, such as small interfering RNA (siRNA) and antisense oligonucleotides (ASO). Therefore, novel therapeutic approaches are widely needed to deliver silencing moieties with maximal trans- fection efficiency and minimal toxicity. This thesis explored the use of mild hyperthermia me- diated by gold nanoparticles (AuNPs) or magnetic nanoparticles (MNPs) to enhance nucleic acids delivery with spatiotemporal control over laser irradiation or magnetic modulation, re- spectively. First, the photothermal effect of AuNPs under visible light irradiation was verified, leading to enhanced cellular uptake. The potential of mild photothermy via AuNPs was demonstrated by effectively silencing the GFP gene in colorectal carcinoma cell line (HCT116) and breast adenocarcinoma cell line (MCF-7), with comparable gene silencing efficiency to commercial transfection reagent, but without cytotoxicity. Improving gene silencing strategies in 3D cell cultures is important since it provides in vitro models that closely resemble the in vivo tumor microenvironment (TME). Then, it was shown that mild photothermy mediated by AuNPs functionalized with ASO decreases c-MYC oncogene expression in HCT116 cells and 7-day spheroids, respectively. Localized magnetic hyperthermia mediated by immobilized MNPs on the cell mem- brane through bioorthogonal chemistry improves the transfection of anti-GFP in MCF-7 cells, with similar efficacy and less cytotoxicity compared to standard transfection reagent. Taking advantage of this approach, an effective IDO1 gene silencing was obtained through the trans- fection of siRNA in dendritic cells derived from an acute monocytic leukemia cell line (THP- 1). Moreover, the upregulation of pro-inflammatory cytokines IL6, TNFA, and IL12 genes and the downregulation of anti-inflammatory IL10 gene might contribute to a more immunogenic state in a TME context. In summary, this thesis highlights nanoparticle-mediated mild hyperthermia as a promising strategy for controlled and efficient nucleic acids delivery that might pave the way for improved gene therapy applications in more complex cancer models. |
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| Autores principais: | Ferreira, Daniela Filipa Cardoso |
| Assunto: | cancer therapy nucleic acids transfection gene Silencing mild hyperthermia gold nanoparticles magnetic nanoparticles |
| Ano: | 2025 |
| País: | Portugal |
| Tipo de documento: | tese de doutoramento |
| Tipo de acesso: | acesso aberto |
| Instituição associada: | Universidade Nova de Lisboa |
| Idioma: | inglês |
| Origem: | Repositório Institucional da UNL |
| Resumo: | Gene therapy relies on the precise transfection of nucleic acid effectors into cancer cells, such as small interfering RNA (siRNA) and antisense oligonucleotides (ASO). Therefore, novel therapeutic approaches are widely needed to deliver silencing moieties with maximal trans- fection efficiency and minimal toxicity. This thesis explored the use of mild hyperthermia me- diated by gold nanoparticles (AuNPs) or magnetic nanoparticles (MNPs) to enhance nucleic acids delivery with spatiotemporal control over laser irradiation or magnetic modulation, re- spectively. First, the photothermal effect of AuNPs under visible light irradiation was verified, leading to enhanced cellular uptake. The potential of mild photothermy via AuNPs was demonstrated by effectively silencing the GFP gene in colorectal carcinoma cell line (HCT116) and breast adenocarcinoma cell line (MCF-7), with comparable gene silencing efficiency to commercial transfection reagent, but without cytotoxicity. Improving gene silencing strategies in 3D cell cultures is important since it provides in vitro models that closely resemble the in vivo tumor microenvironment (TME). Then, it was shown that mild photothermy mediated by AuNPs functionalized with ASO decreases c-MYC oncogene expression in HCT116 cells and 7-day spheroids, respectively. Localized magnetic hyperthermia mediated by immobilized MNPs on the cell mem- brane through bioorthogonal chemistry improves the transfection of anti-GFP in MCF-7 cells, with similar efficacy and less cytotoxicity compared to standard transfection reagent. Taking advantage of this approach, an effective IDO1 gene silencing was obtained through the trans- fection of siRNA in dendritic cells derived from an acute monocytic leukemia cell line (THP- 1). Moreover, the upregulation of pro-inflammatory cytokines IL6, TNFA, and IL12 genes and the downregulation of anti-inflammatory IL10 gene might contribute to a more immunogenic state in a TME context. In summary, this thesis highlights nanoparticle-mediated mild hyperthermia as a promising strategy for controlled and efficient nucleic acids delivery that might pave the way for improved gene therapy applications in more complex cancer models. |
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