Cellular Uptake Mechanism of Nucleic Acid Nanocapsules and Their DNA-Surfactant Building Blocks

Nucleic acid nanocapsules (NANs) are enzyme-responsive DNA-functionalized micelles built for the controlled release of DNA-surfactant conjugates (DSCs) that present sequences with demonstrated therapeutic potential. Here, we investigate the mechanisms by which DSCs gain access to intracellular space...

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Bibliographic Details
Published in:	Bioconjugate chemistry Vol. 34; no. 6; pp. 1004 - 1013
Main Authors:	Pal, Suman, de la Fuente, Ina F., Sawant, Shraddha S., Cannata, Jenna N., He, Wu, Rouge, Jessica L.
Format:	Journal Article
Language:	English
Published:	United States American Chemical Society 21-06-2023
Subjects:	Biological Transport Caveolae Clathrin Controlled release Deoxyribonucleic acid DNA DNA - pharmacology Endocytosis Enzymes External stimuli Flow cytometry Gene sequencing Internalization Micelles Nanocapsules Nanomaterials Nanotechnology Nucleic Acids Nucleotide sequence Receptors Scavenger receptors Surface-Active Agents Surfactants
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Summary:	Nucleic acid nanocapsules (NANs) are enzyme-responsive DNA-functionalized micelles built for the controlled release of DNA-surfactant conjugates (DSCs) that present sequences with demonstrated therapeutic potential. Here, we investigate the mechanisms by which DSCs gain access to intracellular space in vitro and determine the effects of serum on the overall uptake and internalization mechanism of NANs. Using pharmacological inhibitors to selectively block certain pathways, we show, through confocal visualization of cellular distribution and flow cytometry quantification of total cellular association, that scavenger receptor-mediated, caveolae-dependent endocytosis is the major cellular uptake pathway of NANs in the presence and absence of serum. Furthermore, as NANs can be triggered to release DSCs by external stimuli such as enzymes, we sought to examine the uptake profile of particles degraded by enzymes prior to cell-based assays. We found that while scavenger receptor-mediated, caveolae-dependent endocytosis is still at play, energy-independent pathways as well as clathrin-mediated endocytosis are also involved. Overall, this study has helped to elucidate early steps in the cytosolic delivery and therapeutic activity of DSCs packaged into a micellular NAN platform while shedding light on the way in which DNA functionalized nanomaterials in general can be trafficked into cells both as nanostructures and as molecular entities. Importantly, our study also shows that the NAN design in particular is able to stabilize nucleic acids when delivered in the presence of serum, a critical step for effective therapeutic nucleic acid delivery.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 S.P. and I.F.d.l.F. contributed equally to this manuscript.
ISSN:	1043-1802 1520-4812 1520-4812
DOI:	10.1021/acs.bioconjchem.3c00104