Publicação

In vitro transcribed mRNA optimization for gene therapy in the eye

Ver documento

Detalhes bibliográficos
Resumo:Inherited retinal diseases (IRDs) represent a heterogeneous group of degenerative diseases, caused by around 300 gene mutations, without an effective treatment. IRDs constitute the leading cause of blindness worldwide, with a prevalence of 1 in 2000 individuals. Gene therapy emerges as a promis- ing therapeutic option for the possible treatment of IRDs. Recent advancements in mRNA therapeutics motivated the exploration of in vitro transcribed mRNA for therapeutic applications including the COVID- 19 vaccines. Despite the success of mRNA-based vaccination, little research has been done on the application of mRNA technology for ocular diseases. Delivering mRNA to the retina holds substantial advantages, as mRNA is completely functional in the cytoplasm and does not require nuclear entry, hence does not integrate with the genome. Furthermore, it presents a transient nature, low-cost produc- tion, and low immunogenicity. This project aims to explore mRNA efficiency in two retinal pigment epi- thelium (RPE) cell models: ARPE-19, human induced Pluripotent Stem Cells-derived RPE (hiPSCs- RPE) and also retinal organoids. In this work, the delivery of a mRNA (GFPmRNA) using commercial lipid-based nanoparticles (LNPs) was explored. Results demonstrated that mRNA is efficiently delivered to RPE in a short time (1h-3h). Moreover, mRNA complexed with LNPs, successfully transfect both ARPE-19 and hiPSCs- RPE, that lasted up to 30 days without toxicity. Notably, complexed mRNA demonstrated superior transfection efficiency compared to pDNA and AAV2 strategies. On the other hand, AAV6 exhibited similar efficiency compared to mRNA delivery in hiPSCs-RPE and higher in ARPE-19. Despite this, the increased expression of MCP1 was only observed in viral delivery compared to mRNA transfection, suggesting potential immunogenicity. Furthermore, in retinal organoids only few Müller glia cells were transfected. In summary, this study showed a clear advantage of mRNA delivery compared to other strate- gies in RPE cells and highlights the potential for developing mRNA-based gene therapies for the treatment of IRDs.
Autores principais:Castro, Mariana Parreiras Dias Urbano de
Assunto:lARPE-19 Gene therapy human induced Pluripotent Stem Cells mRNA Lipid Nanoparticles Retinal Organoids
Ano:2023
País:Portugal
Tipo de documento:dissertação de mestrado
Tipo de acesso:acesso embargado
Instituição associada:Universidade Nova de Lisboa
Idioma:inglês
Origem:Repositório Institucional da UNL

Registos relacionados

Registos relacionados