Publicação
Application of antisense therapy to control Nakaseomyces glabratus infections
| Resumo: | Canclicla infections affect billions of people worlclwicle, with high morbidity ancl mortality rates (40-60CX,) [l] ancl require long hospital stays ancl increase healthcare costs, which places a significant socio-economic burclen on the economy [2]. Canclicla glabrata (now reclassified as Nakaseomyces glabratus) poses a significant clinical challenge, clue to its increasing levels of resistance to conventional anti fungal therapies, particularly azoles [3]. In this context, antisense therapy (AST) has emergecl as a promising approach in recent clecacles, demonstrating significant potential not only for treating human genetic diseases but also for combating infections [4]. This work aims to clevelop antisense oligonucleoticles (ASOs) targeting resistance-associated genes in N. glabratus, with the objective of restoring susceptibility to conventional antifungal treatments. Specifically, an ASO was clesignecl to target the PDRl gene, a crucial regulator of efflux pump expression ancl membrane permeability, which plays a significant role in antifungal resistance. The therapeutic potential of the anti-PDRl ASO was initially assessed in vitro to determine its effectiveness in reducing gene expression ancl enhancing susceptibility to fluconazole. To enhance bioavailability ancl stability within a living organism, the anti-PDRl ASO was encapsulated in lipicl nanoparticles (LPXs-ASOs), a delivery approach previously clemonstratecl to be effective for other Canclicla species [S]. Subsequently, the in vivo efficacy of the encapsulated ASO was valiclatecl using Galleria mellonella larvae as the moclel organism [6]. In vitro results showecl that the anti-PDRJ ASO was able to effectively recluce the PDRI expression by 40%, even uncler fluconazole-inclucecl conclitions. Aclclitionally, the ASO enhanced the growth inhibition of N. gla/Jratus by at least 20%, both in the presence ancl absence of fluconazole, ancl when ASO was free or encapsulatecl in LPXs. In vivo, the administration of LPXs-ASO significantly enhanced the survival rate ancl health inclex of the infectecl Galleria mellonella larvae moclel by approximately 20'Yc,. Overall, these finclings highlight AST as a promising strategy to resensitize N. gla/Jratus to traditional antifungal clrugs, provicling a new way of aclclressing the urgent issue of antifungal resistance. |
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| Autores principais: | Serra, Patrícia |
| Outros Autores: | Gonçalves, Bruna; Henriques, Mariana; Castro, Joana; Araújo, Daniela; Silva, Sónia Carina |
| Assunto: | Canc/icla infections Antifungal resistance Antisense oligonucleotides Lipid nanoparticles |
| Ano: | 2025 |
| País: | Portugal |
| Tipo de documento: | outro |
| Tipo de acesso: | acesso aberto |
| Instituição associada: | Universidade do Minho |
| Idioma: | inglês |
| Origem: | RepositóriUM - Universidade do Minho |
| Resumo: | Canclicla infections affect billions of people worlclwicle, with high morbidity ancl mortality rates (40-60CX,) [l] ancl require long hospital stays ancl increase healthcare costs, which places a significant socio-economic burclen on the economy [2]. Canclicla glabrata (now reclassified as Nakaseomyces glabratus) poses a significant clinical challenge, clue to its increasing levels of resistance to conventional anti fungal therapies, particularly azoles [3]. In this context, antisense therapy (AST) has emergecl as a promising approach in recent clecacles, demonstrating significant potential not only for treating human genetic diseases but also for combating infections [4]. This work aims to clevelop antisense oligonucleoticles (ASOs) targeting resistance-associated genes in N. glabratus, with the objective of restoring susceptibility to conventional antifungal treatments. Specifically, an ASO was clesignecl to target the PDRl gene, a crucial regulator of efflux pump expression ancl membrane permeability, which plays a significant role in antifungal resistance. The therapeutic potential of the anti-PDRl ASO was initially assessed in vitro to determine its effectiveness in reducing gene expression ancl enhancing susceptibility to fluconazole. To enhance bioavailability ancl stability within a living organism, the anti-PDRl ASO was encapsulated in lipicl nanoparticles (LPXs-ASOs), a delivery approach previously clemonstratecl to be effective for other Canclicla species [S]. Subsequently, the in vivo efficacy of the encapsulated ASO was valiclatecl using Galleria mellonella larvae as the moclel organism [6]. In vitro results showecl that the anti-PDRJ ASO was able to effectively recluce the PDRI expression by 40%, even uncler fluconazole-inclucecl conclitions. Aclclitionally, the ASO enhanced the growth inhibition of N. gla/Jratus by at least 20%, both in the presence ancl absence of fluconazole, ancl when ASO was free or encapsulatecl in LPXs. In vivo, the administration of LPXs-ASO significantly enhanced the survival rate ancl health inclex of the infectecl Galleria mellonella larvae moclel by approximately 20'Yc,. Overall, these finclings highlight AST as a promising strategy to resensitize N. gla/Jratus to traditional antifungal clrugs, provicling a new way of aclclressing the urgent issue of antifungal resistance. |
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