Enhancing RNA-lipid nanoparticle delivery: Organ- and cell-specificity and barcoding strategies

Journal of Controlled Release, Journal Year: 2024, Volume and Issue: 375, P. 366 - 388

Published: Sept. 18, 2024

Recent advancements in RNA therapeutics highlight the critical need for precision gene delivery systems that target specific organs and cells. Lipid nanoparticles (LNPs) have emerged as key vectors delivering mRNA siRNA, offering protection against enzymatic degradation, enabling targeted cellular uptake, facilitating cargo release into cytosol. This review discusses development optimization of organ- cell-specific LNPs, focusing on their design, mechanisms action, therapeutic applications. We explore innovations such DNA/RNA barcoding, which facilitates high-throughput screening precise adjustments formulations. address major challenges, including improving endosomal escape, minimizing off-target effects, enhancing efficiencies. Notable clinical trials recent FDA approvals illustrate practical applications future potential LNP-based therapies. Our findings suggest while considerable progress has been made, continued research is essential to resolve existing limitations bridge gap between pre-clinical evaluation safety efficacy therapeutics. highlights dynamic LNP research. It outlines a roadmap RNA-based medicine.

Language: Английский

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Kinetics of RNA-LNP delivery and protein expression

European Journal of Pharmaceutics and Biopharmaceutics, Journal Year: 2024, Volume and Issue: 197, P. 114222 - 114222

Published: Feb. 20, 2024

Lipid nanoparticles (LNPs) employing ionizable lipids are the most advanced technology for delivery of RNA, notably mRNA, to cells. LNPs represent well-defined core–shell particles with efficient nucleic acid encapsulation, low immunogenicity and enhanced efficacy. While much is known about structure activity LNPs, less attention given timing LNP uptake, cytosolic transfer protein expression. However, kinetics a key factor determining efficiency. Hence quantitative insight into multi-cascaded pathway interest elucidate mechanism delivery. Here, we review experiments as well theoretical modeling mRNA-release We describe sequence stochastic processes mathematical model subsequent translation from mRNA. compile probabilities numbers obtained time resolved microscopy. Specifically, live-cell imaging on single cell arrays (LISCA) allows high-throughput acquisition thousands individual GFP reporter expression courses. The traces yield distribution mRNA life-times, rates onset. Correlation analysis reveals an inverse dependence gene efficiency transfection onset-times. Finally, discuss why release critical in context codelivery multiple species case co-expression or CRISPR/Cas editing.

Language: Английский

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Investigation of Cubosome Interactions with Liposomal Membranes Based on Time-Resolved Small-Angle X-ray Scattering and Laurdan Fluorescence Spectroscopy

The Journal of Physical Chemistry B, Journal Year: 2025, Volume and Issue: 129(9), P. 2461 - 2470

Published: Feb. 25, 2025

Nanosized dispersions of the bicontinuous cubic phase (cubosomes) are emerging carriers for drug delivery. These particles possess well-defined internal structures composed highly-curved lipid bilayers that can accommodate significant payloads. Although cubosomes present promising potential delivery, their physicochemical properties and interactions with cell membranes have not yet been fully understood. To clarify membranes, we investigated changes in structural monoolein (MO) when mixed model at different states using time-resolved small-angle X-ray scattering (TR-SAXS), cryogenic transmission electron microscopy (cryo-TEM), fluorescence spectroscopy. TR-SAXS results showed gradually transitioned from Im3m to lamellar after interacting liposomes. The time change was influenced by fluidity liposome bilayers. Mixing fluid membrane liposomes required less transition vice versa. Cryo-TEM images structure disappeared, leaving behind vesicles interaction, further confirming results. Laurdan probe used assess polarity occurring both during interaction. Examination normalized intensity probed decreasing intensity, followed a recovery which could indicate disintegration formation membrane. Also, kinetics did seem be composition liposomes, line SAXS Assessing generalized polarization (GP340) values mixing revealed hydration resemble counterpart, exchange material between particles. Over time, equilibrated toward an intermediate state two. needed reach final more particularly difference GP340 state. highlight importance examination conditions, such as polarity, implications on interaction liposomal membranes.

Language: Английский

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Small-angle X-ray and neutron scattering applied to lipid-based nanoparticles: Recent advancements across different length scales

Advances in Colloid and Interface Science, Journal Year: 2024, Volume and Issue: 327, P. 103156 - 103156

Published: April 12, 2024

Lipid-based nanoparticles (LNPs), ranging from nanovesicles to non-lamellar assemblies, have gained significant attention in recent years, as versatile carriers for delivering drugs, vaccines, and nutrients. Small-angle scattering methods, employing X-rays (SAXS) or neutrons (SANS), represent unique tools unveil structure, dynamics, interactions of such particles on different length scales, spanning the nano molecular scale. This review explores state-of-the-art methods applied structure lipid-based their with drugs bioactive molecules, inform rational design formulation medical applications. We will focus complementary information accessible neutrons, insights colloidal processes at a nanoscale level details lipid organization LNPs (SANS). In addition, we new opportunities offered by Time-resolved (TR)-SAXS -SANS investigation dynamic involving LNPs. These span real-time monitoring structural evolution response endogenous external stimuli (TR-SANS), kinetics diffusion exchange upon interaction biomolecules (TR-SANS). Finally, spotlight novel combinations SAXS SANS on-line techniques, recently enabled Large Scale Facilities neutrons. emerging technology enables synchronized multi-method investigation, offering exciting simultaneous characterization chemical mechanical properties

Language: Английский

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Extracellular vesicles versus lipid nanoparticles for the delivery of nucleic acids

Advanced Drug Delivery Reviews, Journal Year: 2024, Volume and Issue: unknown, P. 115461 - 115461

Published: Oct. 1, 2024

Language: Английский

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Quantitative Visualization of Lipid Nanoparticle Fusion as a Function of Formulation and Process Parameters

ACS Nano, Journal Year: 2024, Volume and Issue: 18(28), P. 18191 - 18201

Published: July 5, 2024

Lipid nanoparticles (LNPs) have proven to be promising delivery vehicles for RNA-based vaccines and therapeutics, particularly in LNP formulations containing ionizable cationic lipids that undergo protonation/deprotonation response buffer pH changes. These are typically formulated using a rapid mixing technique at low pH, followed by return physiological triggers LNP-LNP fusion. A detailed understanding of these dynamic processes is crucial optimize the overall performance efficiency LNPs. However, knowledge gaps persist regarding how particle formation mechanisms impact drug loading functions. In this work, we employ single-molecule Convex Lens-induced Confinement (CLiC) microscopy combination with Förster resonance energy transfer (FRET) measurements study fusion dynamics relation various formulation parameters, including lipid concentration, conditions, ratio, PEG-lipid concentrations, selection. Our results reveal strong correlation between measured parameters used; findings consistent DLS Cryo-TEM-based assays. offer cost-effective method characterizing screening potential candidates can provide additional insights into their design, opportunities optimization.

Language: Английский

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Inverse Cubic and Hexagonal Mesophase Evolution within Ionizable Lipid Nanoparticles Correlates with mRNA Transfection in Macrophages

Journal of the American Chemical Society, Journal Year: 2023, Volume and Issue: unknown

Published: Oct. 23, 2023

mRNA lipid nanoparticle (LNP) technology presents enormous opportunities to prevent and treat various diseases. Here, we developed a novel series of LNPs containing ionizable amino-lipids showing remarkable array tunable pH-sensitive lyotropic liquid crystalline mesophases including the inverse bicontinuous cubic hexagonal phases characterized by high-throughput synchrotron radiation X-ray scattering. Furthermore, with an interest in developing therapeutics for lung macrophage targeting, discovered that there is strong correlation between mesophase transition during acidification association/transfection efficiency mRNAs. The slight molecular structural differences SM-102 ALC-0315 lipids are linked LNP's ability transform their internal structures from amorphous state micellar, hexagonal, finally endosomal maturation. showed exceptionally improved transfection due form structure at lower pH than analogues, which remained within structure, previously attributed promoting escape LNPs. Overall, new knowledge draws our attention important role escape, LNP libraries reported herein have broad prospects advancing therapeutics.

Language: Английский

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Stable and inhalable powder formulation of mRNA-LNPs using pH-modified spray-freeze drying

International Journal of Pharmaceutics, Journal Year: 2024, Volume and Issue: 665, P. 124632 - 124632

Published: Sept. 7, 2024

Language: Английский

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Ether bond-modified lipid nanoparticles for enhancing the treatment effect of hepatic fibrosis

International Journal of Pharmaceutics, Journal Year: 2025, Volume and Issue: 671, P. 125192 - 125192

Published: Jan. 16, 2025

Language: Английский

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Choice of organic solvent affects function of mRNA-LNP; pyridine produces highly functional mRNA-LNP

International Journal of Pharmaceutics, Journal Year: 2025, Volume and Issue: unknown, P. 125367 - 125367

Published: Feb. 1, 2025

Language: Английский

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