TitleCholesterol Sequestration from Caveolae/Lipid Rafts Enhances Cationic Liposome-Mediated Nucleic Acid Delivery into Endothelial Cells.
Publication TypeJournal Article
Year of Publication2021
AuthorsMaddila SChandar, Voshavar C, Arjunan P, Chowath RPrakash, Rachamalla HKrishna Re, Balakrishnan B, Balasubramanian P, Banerjee R, Marepally S
Date Published2021 Jul 30
KeywordsAnimals, Caveolae, Caveolin 1, Cell Line, Transformed, Cholesterol, Clathrin, DNA, Endocytosis, Endothelial Cells, Filipin, Gene Expression, Liposomes, Membrane Microdomains, Nystatin, Phosphatidylethanolamines, Pinocytosis, Plasmids, Rats, RNA, Small Interfering, Transfection

Delivering nucleic acids into the endothelium has great potential in treating vascular diseases. However, endothelial cells, which line the vasculature, are considered as sensitive in nature and hard to transfect. Low transfection efficacies in endothelial cells limit their potential therapeutic applications. Towards improving the transfection efficiency, we made an effort to understand the internalization of lipoplexes into the cells, which is the first and most critical step in nucleic acid transfections. In this study, we demonstrated that the transient modulation of caveolae/lipid rafts mediated endocytosis with the cholesterol-sequestrating agents, nystatin, filipin III, and siRNA against Cav-1, which significantly increased the transfection properties of cationic lipid-(2-hydroxy--methyl-,-bis(2-tetradecanamidoethyl)ethanaminium chloride), namely, amide liposomes in combination with 1,2-Dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) (AD Liposomes) in liver sinusoidal endothelial cells (SK-Hep1). In particular, nystatin was found to be highly effective with 2-3-fold enhanced transfection efficacy when compared with amide liposomes in combination with Cholesterol (AC), by switching lipoplex internalization predominantly through clathrin-mediated endocytosis and macropinocytosis.

Alternate JournalMolecules
PubMed ID34361779
PubMed Central IDPMC8346983
Grant ListU54 MD007582 / MD / NIMHD NIH HHS / United States
BT/PR25841/GET/119/162/2017 / / Department of Biotechnology, Ministry of Science and Technology, India /