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Heat up, silence on
Publication . Ferreira, Daniela; Asín, Laura; Idiago-López, Javier; Grazú, Valeria; Fuente, Jesús M. de la; Fratila, Raluca M.; Baptista, Pedro V.; Fernandes, Alexandra R.; DCV - Departamento de Ciências da Vida; UCIBIO - Applied Molecular Biosciences Unit; Springer Science and Business Media Deutschland GmbH
Dendritic cells (DCs) are well-known antigen-presenting cells which have an important role in cancer immunomodulation due to the effective regulation of immune responses in the tumor microenvironment (TME). Indoleamine 2,3-dioxygenase 1 gene (IDO1) is upregulated in many types of cancers and associated with a poor prognosis, contributing to an immunosuppressive TME. IDO1 silencing in DCs is considered a promising new strategy in gene therapy owing to their capability to regulate T cells function and activation. This study focuses on the use of magnetic hyperthermia (MH) combined with bioorthogonal chemistry to promote siRNA transfection against IDO1 in THP-1-derived DCs. Magnetic nanoparticles (MNPs) functionalized with cyclooctyne moieties were attached by strain-promoted azide-alkyne cycloaddition to DCs membranes engineered to express artificial azide receptors. Upon the application of an alternating magnetic field, the MNPs generate heat and trigger the thermal disruption of the cell membrane. Results show that IDO1 gene expression decreases around 70% in THP-1-derived DCs, and that the MH-promoted transfection presents a silencing effect comparable to that attained with a gold standard Lipofectamine reagent, but with less cytotoxicity. Additionally, IDO1 silencing promotes the upregulation of mRNA levels of pro-inflammatory cytokines IL-6, TNF-α and IL-12A, and the downregulation of anti-inflammatory cytokine IL-10, providing a more immunogenic state which may lead to THP-1-derived DCs activation for future T cells antitumor response. Our findings reveal the potential of MH-mediated transfection to enhance the intracellular delivery of silencing moieties in cells difficult to transfect, such as DCs, as well as demonstrate the possibility of silencing IDO1 gene to overcome the immunosuppressive barrier imposed by the TME for cancer therapy.
Gold nanoparticles for vectorization of nucleic acids for cancer therapeutics
Publication . Ferreira, Daniela; Fontinha, David; Martins, Catarina; Pires, David; Fernandes, Alexandra R.; Baptista, Pedro V.; DCV - Departamento de Ciências da Vida; UCIBIO - Applied Molecular Biosciences Unit; MDPI - Multidisciplinary Digital Publishing Institute
Cancer remains a complex medical challenge and one of the leading causes of death worldwide. Nanomedicines have been proposed as innovative platforms to tackle these complex diseases, where the combination of several treatment strategies might enhance therapy success. Among these nanomedicines, nanoparticle mediated delivery of nucleic acids has been put forward as key instrument to modulate gene expression, be it targeted gene silencing, interference RNA mechanisms and/or gene edition. These novel delivery systems have strongly relied on nanoparticles and, in particular, gold nanoparticles (AuNPs) have paved the way for efficient delivery systems due to the possibility to fine-tune their size, shape and surface properties, coupled to the ease of functionalization with different biomolecules. Herein, we shall address the different molecular tools for modulation of expression of oncogenes and tumor suppressor genes and discuss the state-of-the-art of AuNP functionalization for nucleic acid delivery both in vitro and in vivo models. Furthermore, we shall highlight the clinical applications of these spherical AuNP based conjugates for gene delivery, current challenges, and future perspectives in nanomedicine.
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Fundação para a Ciência e a Tecnologia
Programa de financiamento
3599-PPCDT
Número da atribuição
M-ERA-NET2/0008/2016