RT Journal Article
T1 Surfactant Protein B promotes cytosolic SiRNA delivery by adopting a Virus-like mechanism of action
A1 Guagliardo, Roberta
A1 Herman, Lore
A1 Penders, Jelle
A1 Zamborlin, Agata
A1 Keersmaecker, Herlinde de
A1 Van de Vyver, Thijs
A1 Verstraeten, Sandrine
A1 Merckx, Pieterjan
A1 Mingeot-Leclercq, Marie-Paule
A1 Echaide Torreguitar, Mercedes
A1 Pérez Gil, Jesús
A1 Stevens, Molly M.
A1 De Smedt, Stefaan
A1 Raemdonck, Koen
AB RNA therapeutics are poised to revolutionize medicine. To unlock the full potential of RNA drugs, safe and efficient (nano)formulations to deliver them inside target cells are required. Endosomal sequestration of nanocarriers represents a major bottleneck in nucleic acid delivery. Gaining more detailed information on the intracellular behavior of RNA nanocarriers is crucial to rationally develop delivery systems with improved therapeutic efficiency. Surfactant protein B (SPB) is a key component of pulmonary surfactant (PS), essential for mammalian breathing. In contrast to the general belief that PS should be regarded as a barrier for inhaled nanomedicines, we recently discovered the ability of SP-B to promote gene silencing by siRNA-loaded and lipid-coated nanogels. However, the mechanisms governing this process are poorly understood. The major objective of this work was to obtain mechanistic insights into the SP-B-mediated cellular delivery of siRNA. To this end, we combined siRNA knockdown experiments, confocal microscopy, and focused ion beam scanning electron microscopy imaging in an in vitro non-small-cell lung carcinoma model with lipid mixing assays on vesicles that mimic the composition of (intra)cellular membranes. Our work highlights a strong correlation between SP-B-mediated fusion with anionic endosomal membranes and cytosolic siRNA delivery, a mode of action resembling that of certain viruses and virus-derived cell-penetrating peptides. Building on these gained insights, we optimized the SP-B proteolipid composition, which dramatically improved delivery efficiency. Altogether, our work provides a mechanistic understanding of SP-B-induced perturbation of intracellular membranes, offering opportunities to fuel the rational design of SP-B-inspired RNA nanoformulations for inhalation therapy.
PB American Chemical Society
SN 1936-0851; Electronic: 1936-086X
YR 2021
FD 2021-03-16
LK https://hdl.handle.net/20.500.14352/8462
UL https://hdl.handle.net/20.500.14352/8462
LA eng
NO Unión Europea. Horizonte 2020
NO Ministerio de Ciencia e Innovación ((MICINN)
NO Comunidad de Madrid
NO Ghent University Special Research Fund
NO Research Foundation Flanders (Belgium)
NO Belgian Fonds de la Recherche Scientifique
NO Action de Recherche Concertée (ARC)
NO Flanders Innovation & Entrepreneurship Agency
DS Docta Complutense
RD 13 abr 2025