scholarly journals Illuminating endosomal escape of polymorphic lipid nanoparticles that boost mRNA delivery

2021 ◽  
Author(s):  
Marco Herrera ◽  
Jeonghwan Kim ◽  
Yulia Eygeris ◽  
Antony Jozic ◽  
Gaurav Sahay
Keyword(s):  
Lipid Nanoparticles ◽  
Endosomal Escape

Galectin8-GFP cytosolic redistribution to bright puncta serve as sensor for LNP escape from endosomal compartments.

scholarly journals Cytosolic protein delivery using pH-responsive, charge-reversible lipid nanoparticles

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Yusuke Hirai ◽  
Hisaaki Hirose ◽  
Miki Imanishi ◽  
Tomohiro Asai ◽  
Shiroh Futaki
Keyword(s):  
Protein Delivery ◽  
Electrostatic Interactions ◽  
Fluorescent Protein ◽  
Lipid Nanoparticles ◽  
Endosomal Escape ◽  
Ph Responsive ◽  
Cargo Proteins ◽  
Green Fluorescent ◽  
Intracellular Protein Delivery ◽  
Endosomal Release

AbstractAlthough proteins have attractive features as biopharmaceuticals, the difficulty in delivering them into the cell interior limits their applicability. Lipid nanoparticles (LNPs) are a promising class of delivery vehicles. When designing a protein delivery system based on LNPs, the major challenges include: (i) formulation of LNPs with defined particle sizes and dispersity, (ii) efficient encapsulation of cargo proteins into LNPs, and (iii) effective cellular uptake and endosomal release into the cytosol. Dioleoylglycerophosphate-diethylenediamine (DOP-DEDA) is a pH-responsive, charge-reversible lipid. The aim of this study was to evaluate the applicability of DOP-DEDA-based LNPs for intracellular protein delivery. Considering the importance of electrostatic interactions in protein encapsulation into LNPs, a negatively charged green fluorescent protein (GFP) analog was successfully encapsulated into DOP-DEDA-based LNPs to yield diameters and polydispersity index of < 200 nm and < 0.2, respectively. Moreover, ~ 80% of the cargo proteins was encapsulated into the LNPs. Cytosolic distribution of fluorescent signals of the protein was observed for up to ~ 90% cells treated with the LNPs, indicating the facilitated endocytic uptake and endosomal escape of the cargo attained using the LNP system.

scholarly journals Linkage between endosomal escape of LNP-mRNA and loading into EVs for transport to other cells

2019 ◽  
Vol 10 (1) ◽  
Author(s):  
Marco Maugeri ◽  
Muhammad Nawaz ◽  
Alexandros Papadimitriou ◽  
Annelie Angerfors ◽  
Alessandro Camponeschi ◽  
...  
Keyword(s):  
Inflammatory Cytokines ◽  
Lipid Nanoparticles ◽  
Endosomal Escape ◽  
Molar Ratio ◽  
Great Promise ◽  
Human Erythropoietin ◽  
First Time ◽  
In Vivo Delivery ◽  
Lower Expression

Abstract RNA-based therapeutics hold great promise for treating diseases and lipid nanoparticles (LNPs) represent the most advanced platform for RNA delivery. However, the fate of the LNP-mRNA after endosome-engulfing and escape from the autophagy-lysosomal pathway remains unclear. To investigate this, mRNA (encoding human erythropoietin) was delivered to cells using LNPs, which shows, for the first time, a link between LNP-mRNA endocytosis and its packaging into extracellular vesicles (endo-EVs: secreted after the endocytosis of LNP-mRNA). Endosomal escape of LNP-mRNA is dependent on the molar ratio between ionizable lipids and mRNA nucleotides. Our results show that fractions of ionizable lipids and mRNA (1:1 molar ratio of hEPO mRNA nucleotides:ionizable lipids) of endocytosed LNPs were detected in endo-EVs. Importantly, these EVs can protect the exogenous mRNA during in vivo delivery to produce human protein in mice, detected in plasma and organs. Compared to LNPs, endo-EVs cause lower expression of inflammatory cytokines.

scholarly journals Endosomal escape of delivered mRNA from endosomal recycling tubules visualized at the nanoscale

2021 ◽  
Vol 221 (2) ◽  
Author(s):  
Prasath Paramasivam ◽  
Christian Franke ◽  
Martin Stöter ◽  
Andreas Höijer ◽  
Stefano Bartesaghi ◽  
...  
Keyword(s):  
Super Resolution ◽  
Lipid Nanoparticles ◽  
Endosomal Escape ◽  
Human Adipocytes ◽  
Promising Strategy ◽  
Cargo Transport ◽  
Endosomal Recycling ◽  
Endosomal Acidification ◽  
Poor Understanding ◽  
Super Resolution Microscopy

Delivery of exogenous mRNA using lipid nanoparticles (LNPs) is a promising strategy for therapeutics. However, a bottleneck remains in the poor understanding of the parameters that correlate with endosomal escape versus cytotoxicity. To address this problem, we compared the endosomal distribution of six LNP-mRNA formulations of diverse chemical composition and efficacy, similar to those used in mRNA-based vaccines, in primary human adipocytes, fibroblasts, and HeLa cells. Surprisingly, we found that total uptake is not a sufficient predictor of delivery, and different LNPs vary considerably in endosomal distributions. Prolonged uptake impaired endosomal acidification, a sign of cytotoxicity, and caused mRNA to accumulate in compartments defective in cargo transport and unproductive for delivery. In contrast, early endocytic/recycling compartments have the highest probability for mRNA escape. By using super-resolution microscopy, we could resolve a single LNP-mRNA within subendosomal compartments and capture events of mRNA escape from endosomal recycling tubules. Our results change the view of the mechanisms of endosomal escape and define quantitative parameters to guide the development of mRNA formulations toward higher efficacy and lower cytotoxicity.

scholarly journals DNA-inspired nanomaterials for enhanced endosomal escape

2021 ◽  
Vol 118 (19) ◽  
pp. e2104511118
Author(s):  
Jinhyung Lee ◽  
Ian Sands ◽  
Wuxia Zhang ◽  
Libo Zhou ◽  
Yupeng Chen
Keyword(s):  
Noncovalent Interactions ◽  
Lipid Nanoparticles ◽  
Endosomal Escape ◽  
Side Chain ◽  
Amino Acid Side Chain ◽  
Proof Of Concept ◽  
Cationic Polymers ◽  
Rnai Therapeutics ◽  
Delivery Vehicles ◽  
Interfering Rna

To realize RNA interference (RNAi) therapeutics, it is necessary to deliver therapeutic RNAs (such as small interfering RNA or siRNA) into cell cytoplasm. A major challenge of RNAi therapeutics is the endosomal entrapment of the delivered siRNA. In this study, we developed a family of delivery vehicles called Janus base nanopieces (NPs). They are rod-shaped nanoparticles formed by bundles of Janus base nanotubes (JBNTs) with RNA cargoes incorporated inside via charge interactions. JBNTs are formed by noncovalent interactions of small molecules consisting of a base component mimicking DNA bases and an amino acid side chain. NPs presented many advantages over conventional delivery materials. NPs efficiently entered cells via macropinocytosis similar to lipid nanoparticles while presenting much better endosomal escape ability than lipid nanoparticles; NPs escaped from endosomes via a “proton sponge” effect similar to cationic polymers while presenting significant lower cytotoxicity compared to polymers and lipids due to their noncovalent structures and DNA-mimicking chemistry. In a proof-of-concept experiment, we have shown that NPs are promising candidates for antiviral delivery applications, which may be used for conditions such as COVID-19 in the future.

Enhancement of oral bioavailability of pentoxifylline by solid lipid nanoparticles

2009 ◽  
Vol 00 (00) ◽  
pp. 090820062440031-9 ◽  
Author(s):  
Jaleh Varshosaz ◽  
Mohsen Minayian ◽  
Elaheh Moazen
Keyword(s):  
Solid Lipid Nanoparticles ◽  
Oral Bioavailability ◽  
Lipid Nanoparticles ◽  
Solid Lipid

Development and optimization of solid lipid nanoparticles of amikacin by central composite design

2009 ◽  
Vol 00 (00) ◽  
pp. 090721051030036-8
Author(s):  
Jaleh Varshosaz ◽  
Solmaz Ghaffari ◽  
Mohammad Reza Khoshayand ◽  
Fatemeh Atyabi ◽  
Shirzad Azarmi ◽  
...  
Keyword(s):  
Central Composite Design ◽  
Solid Lipid Nanoparticles ◽  
Lipid Nanoparticles ◽  
Solid Lipid ◽  
Central Composite

Solid lipid nanoparticles for oral delivery of curcumin

Planta Medica ◽  
2013 ◽  
Vol 79 (13) ◽  
Author(s):  
C Righeschi ◽  
M Bergonzi ◽  
B Isacchi ◽  
A Bilia
Keyword(s):  
Solid Lipid Nanoparticles ◽  
Oral Delivery ◽  
Lipid Nanoparticles ◽  
Solid Lipid

Development of Solid Lipid Nanoparticles of Lamivudine for Brain Targeting

2009 ◽  
Vol 1 (1) ◽  
Author(s):  
Pravin Patil ◽  
Anil Sharma ◽  
Subhash Dadarwal ◽  
Vijay Sharma
Keyword(s):  
Drug Delivery ◽  
Drug Release ◽  
Solid Lipid Nanoparticles ◽  
Rotation Speed ◽  
Lipid Nanoparticles ◽  
Brain Targeting ◽  
Brain Penetration ◽  
Solid Lipid ◽  
Diffusion Technique

The objective of present investigation was to enhance brain penetration of Lamivudine, one of the most widely used drugs for the treatment of AIDS. This was achieved through incorporating the drug into solid lipid nanoparticles (SLN) prepared by using emulsion solvent diffusion technique. The formulations were characterized for surface morphology, size and size distribution, percent drug entrapment and drug release. The optimum rotation speed, resulting into better drug entrapment and percent yield, was in the range of 1000-1250 r/min. In vitro cumulative % drug release from optimized SLN formulation was found 40-50 % in PBS (pH-7.4) and SGF (pH-1.2) respectively for 10 h. After 24 h more than 65 % of the drug was released from all formulations in both mediums meeting the requirement for drug delivery for prolong period of time.

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