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Formulating and Characterizing Lipid Nanoparticles for Gene Delivery using a Microfluidic Mixing Platform
Published on: February 25, 2021
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PBAE-PEG based lipid nanoparticles for lung cell-specific gene delivery
Biorxiv : the Preprint Server for Biology
|September 4, 2024
Summary
This study developed novel lipid nanoparticles (LNPs) for delivering messenger RNA (mRNA) to lung cells. Different LNP formulations showed cell-specific delivery via intravenous or intratracheal routes, highlighting potential for pulmonary gene therapy.
Area of Science:
- Biotechnology
- Nanomedicine
- Pulmonary Medicine
Background:
- Modified messenger RNA (mRNA) delivered via lipid nanoparticles (LNPs) shows promise for treating genetic and acquired disorders.
- The COVID-19 pandemic highlighted the potential of mRNA-LNP technology.
Purpose of the Study:
- To develop and evaluate PEGylated (PBAE-PEG) and non-PEGylated (PBAE) lipid nanoparticles for mRNA delivery to pulmonary cells in vivo.
- To assess the cell-specific transfection efficiency of different LNP formulations and administration routes in the lungs.
Main Methods:
- Development of PEGylated and non-PEGylated PBAE lipids formulated into LNPs with 4A3-SC8/DOPE/cholesterol/DOTAP.
- In vitro transfection assays using HEK293T and H441 cells.
- In vivo studies in mice involving intravenous and intratracheal administration of LNPs carrying Cre-recombinase mRNA.
- Evaluation of Cre-recombinase expression and recombination in various lung cell types and endothelial cells.
Main Results:
- PBAE-PEG/LNPs demonstrated high transfection efficiency in HEK293T and H441 cells in vitro.
- Intravenous administration of PBAE-PEG/LNPs resulted in high transfection of pulmonary vascular endothelial cells.
- Intratracheal administration led to efficient and selective transfection of lung epithelial cells, including club cells and alveolar type 2 cells.
- PBAE-PEG/LNPs were more effective for alveolar type 2 cells, while PBAE/LNPs favored secretory airway cells after intratracheal delivery.
- LNPs failed to cross the pulmonary endothelial-to-epithelial barrier.
- 5-methoxyuridine modified mRNA showed improved in vivo efficiency compared to unmodified mRNA.
Conclusions:
- Developed LNP formulations enable targeted mRNA delivery to specific pulmonary cell types.
- Administration route dictates cell tropism, with intravenous targeting endothelial cells and intratracheal targeting epithelial cells.
- These findings support the potential of LNP-mediated mRNA delivery for pulmonary gene therapies.
