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@ARTICLE{Philipp:603710,
author = {Philipp, Julian and Dabkowska, Aleksandra and Reiser, Anita
and Frank, Kilian and Krzysztoń, Rafał and Brummer,
Christiane and Nickel, Bert and Blanchet, Clement E. and
Sudarsan, Akhil and Ibrahim, Mohd and Johansson, Svante and
Skantze, Pia and Skantze, Urban and Östman, Sofia and
Johansson, Marie and Henderson, Neil and Elvevold, Kjetil
and Smedsrød, Bård and Schwierz, Nadine and Lindfors,
Lennart and Rädler, Joachim O.},
title = {p{H}-dependent structural transitions in cationic ionizable
lipid mesophases are critical for lipid nanoparticle
function},
journal = {Proceedings of the National Academy of Sciences of the
United States of America},
volume = {120},
number = {50},
issn = {0027-8424},
address = {Washington, DC},
publisher = {National Acad. of Sciences},
reportid = {PUBDB-2024-00898},
pages = {e2310491120},
year = {2023},
abstract = {Lipid nanoparticles (LNPs) are advanced core-shell
particles for messenger RNA (mRNA) based therapies that are
made of polyethylene glycol (PEG) lipid,
distearoylphosphatidylcholine (DSPC), cationic ionizable
lipid (CIL), cholesterol (chol), and mRNA. Yet the mechanism
of pH-dependent response that is believed to cause endosomal
release of LNPs is not well understood. Here, we show that
eGFP (enhanced green fluorescent protein) protein expression
in the mouse liver mediated by the ionizable lipids
DLin-MC3-DMA (MC3), DLin-KC2-DMA (KC2), and DLinDMA (DD)
ranks MC3 ≥ KC2 > DD despite similar delivery of mRNA per
cell in all cell fractions isolated. We hypothesize that the
three CIL-LNPs react differently to pH changes and hence
study the structure of CIL/chol bulk phases in water. Using
synchrotron X-ray scattering a sequence of ordered CIL/chol
mesophases with lowering pH values are observed. These
phases show isotropic inverse micellar, cubic Fd3m inverse
micellar, inverse hexagonal and bicontinuous cubic Pn3m
symmetry. If polyadenylic acid, as mRNA surrogate, is added
to CIL/chol, excess lipid coexists with a condensed nucleic
acid lipidphase. The next-neighbor distance in the excess
phase shows a discontinuity at the Fd3m inverse micellar to
inverse hexagonal transition occurring at pH 6 with
distinctly larger spacing and hydration for DD vs. MC3 and
KC2. In mRNA LNPs, DD showed larger internal spacing, as
well as retarded onset and reduced level of DD-LNP-mediated
eGFP expression in vitro compared to MC3 and KC2. Our data
suggest that the pH-driven Fd3m- transition in bulk phases
is a hallmark of CIL-specific differences in mRNA LNP
efficacy.},
cin = {EMBL-User / EMBL},
ddc = {500},
cid = {I:(DE-H253)EMBL-User-20120814 / I:(DE-H253)EMBL-20120731},
pnm = {6G3 - PETRA III (DESY) (POF4-6G3)},
pid = {G:(DE-HGF)POF4-6G3},
experiment = {EXP:(DE-H253)P-P12-20150101},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:38055742},
UT = {WOS:001498765900001},
doi = {10.1073/pnas.2310491120},
url = {https://bib-pubdb1.desy.de/record/603710},
}