Selective Transfection of a Transferrin Receptor-Expressing Cell Line With DNA-Lipid Nanoparticles and Synthesis of Parasite-Derived Glycans as Biomarkers for Leishmaniasis
Abstract
Despite notable progress in lipid nanoparticle (LNP)-mediated gene delivery, achieving selective transfection of specific cell types, such as cancer cells, remains a significant hurdle, hindering the advancement of innovative gene therapies. In this study, we engineered an LNP formulation encapsulating plasmid DNA (pDNA) encoding the monomeric Green Lantern (mGL) fluorescent reporter protein. The DT7 peptide ligand targeting human transferrin receptor 1 (hTfR1) was also conjugated to the LNP surface for targeted delivery to hTfR1-expressing cells. Optimization of LNP composition yielded favorable particle diameter, ζ-potential, yield, and pDNA encapsulation efficiency. Evaluation of transfection selectivity using a panel of two engineered cell lines, CHO-TRVb-hTfR1 expressing hTfR1 and CHO-TRVb-neo lacking this receptor, was accomplished through flow cytometry, measuring the expression of the fluorescent mGL protein. Our experiments revealed a dose-dependent mGL expression in the evaluated range of LNP concentrations, with up to 20% transfection efficiency in CHO-TRVb-hTfR1 cells and markedly lower efficiency (up to 4%) in CHO-TRVb-neo cells, indicating a five-fold enhanced selectivity for hTfR1-expressing cells. These findings suggest the potential of this LNP platform, or similar ones modified with DT7, for in vivo applications, particularly in the selective targeting of malignant cells overexpressing hTfR1. Further work aims to enhance LNP selectivity, including the development of dual-targeting LNPs and exploring additional ligands for improved therapeutic precision.
Subject Area
Chemistry|Biomedical engineering|Immunology|Genetics
Recommended Citation
Vinales Lozano, Irodiel, "Selective Transfection of a Transferrin Receptor-Expressing Cell Line With DNA-Lipid Nanoparticles and Synthesis of Parasite-Derived Glycans as Biomarkers for Leishmaniasis" (2024). ETD Collection for University of Texas, El Paso. AAI31292522.
https://scholarworks.utep.edu/dissertations/AAI31292522