Loading of Extracellular Vesicles with Chemically Stabilized Hydrophobic  siRNAs for the Treatment of Disease in the Central Nervous System

Haraszti, Reka A.; Coles, Andrew H.; Aronin, Neil; Khvorova, Anastasia; Didiot, Marie-Cécile

doi:10.21769/bioprotoc.2338

Cited by 10 publications

(6 citation statements)

References 28 publications

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“…However, we did not observe in situ assembly of unconjugated hsiRNAs with lipoproteins in mouse serum, suggesting that an entropic driving force, such as a lipid anchor, is required for binding. This constraint, which we and others have described for oligonucleotide loading into purified exosomes (Biscans et al, manuscript in press) [35][36][37] , provides a rationale for why hydrophobicity drives hsiRNA partitioning into distinct lipid transport pathways. VLDL and LDL carry highly-nonpolar saturated triglycerides and unsaturated cholesteryl esters, respectively, while HDL carries cholesterol and unsaturated phospholipids (relatively more polar).…”

Section: Discussionmentioning

confidence: 74%

Hydrophobicity drives the systemic distribution of lipid-conjugated siRNAs via lipid transport pathways

Osborn

Coles

Biscans

et al. 2018

Preprint

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Efficient delivery of therapeutic RNA is the fundamental obstacle preventing its clinical utility. Lipid conjugation improves plasma half-life, tissue accumulation, and cellular uptake of small interfering RNAs (siRNAs). However, the impact of conjugate structure and hydrophobicity on siRNA pharmacokinetics is unclear, impeding the design of clinically relevant lipid-siRNAs. Using a panel of biologically-occurring lipids, we show that lipid conjugation modulates siRNA hydrophobicity and governs spontaneous partitioning into distinct plasma lipoprotein classes in vivo. Lipoprotein binding influences siRNA distribution by delaying renal excretion and promoting uptake into lipoprotein receptor-enriched tissues. Lipid-siRNAs elicit mRNA silencing without causing toxicity in a tissue-specific manner. Lipid-siRNA internalization occurs independently of lipoprotein endocytosis, and is mediated by siRNA phosphorothioate modifications. Although biomimetic lipoprotein nanoparticles have been considered for the enhancement of siRNA delivery, our findings suggest that hydrophobic modifications can be leveraged to incorporate therapeutic siRNA into endogenous lipid transport pathways without the requirement for synthetic formulation. IntroductionFor over a decade, the underlying obstacle preventing the widespread clinical use of small interfering RNA (siRNA)-based therapies has been efficient and safe in vivo delivery. siRNAs are large (~14 kDa), polyanionic macromolecules that require extensive modifications to improve their inherently-poor pharmacological properties (e.g. plasma half-life of <5 min before renal excretion) 1,2 . Lipid-and polymer-based nanoparticles, which are effective transfection agents in vitro, can prolong circulation time and improve stability and bioavailability in vivo. However, nanoparticle delivery is typically limited to clearance organs with fenestrated and/or discontinuous endothelium (e.g. liver, spleen, and certain tumors). Moreover, nanoparticles can interact with opsonin proteins that enhance clearance by macrophage phagocytosis.

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Section: Discussionmentioning

confidence: 74%

Hydrophobicity drives the systemic distribution of lipid-conjugated siRNAs via lipid transport pathways

Osborn

Coles

Biscans

et al. 2018

Preprint

Self Cite

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“…Passive loading represents a non-invasive strategy to co-incubate therapeutic cargo with isolated EVs in a highly scalable manner, which does not involve invasive manipulation or engineering of parent cells or EVs. In many studies, nucleotides chemically modified with cholesterol or similar hydrophobic moieties were co-cultured with isolated EVs in solution at 37°C to passively associate with the EV membrane [ 28 , 136 , 137 ]. This passive affinity hypothesis was based upon naked nucleotide entry into cells via association with cholesterol [ 138 ].…”

Section: Techniques For Loading Evsmentioning

confidence: 99%

Therapeutic Potential of Nucleic Acids when Combined with Extracellular Vesicles

2021

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Extracellular vesicles (EVs), endogenous nanocarriers of proteins, lipids, and genetic material, have been harnessed as intrinsic delivery vectors for nucleic acid therapies. EVs are nanosized lipid bilayer bound vesicles released from most cell types responsible for delivery of functional biologic material to mediate intercellular communication and to modulate recipient cell phenotypes. Due to their innate biological role and composition, EVs possess several advantages as delivery vectors for nucleic acid based therapies including low immunogenicity and toxicity, high bioavailability, and ability to be engineered to enhance targeting to specific recipient cells in vivo. In this review, the current understanding of the biological role of EVs as well as the advancements in loading EVs to deliver nucleic acid therapies are summarized. We discuss the current methods and associated challenges in loading EVs and the prospects of utilizing the inherent characteristics of EVs as a delivery vector of nucleic acid therapies for genetic disorders.

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“…Post-loading can be subdivided into passive loading, such as by physical incubation, and active loading with instances of electroporation or sonication. Furthermore, the functional small RNAs delivery using electroporated EVs has been shown to be a success in several reports but it depends on the small RNA species [58][59][60][61][62]. Usman and colleagues used the electroporation method for post-loading of RNAs into RBCEVs [55].…”

Section: Using Red Blood Cell Extracellular Vesicles (Rbcevs) For Thementioning

confidence: 99%

The Biology and Therapeutic Applications of Red Blood Cell Extracellular Vesicles

Zhang¹,

Kiomourtzis²,

Lam³

et al. 2019

Erythrocyte

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This chapter focuses on the biology of red blood cell extracellular vesicles (RBCEVs) in normal and diseased conditions, and the potential application of RBCEVs in treatment. Extracellular vesicles (EVs) refer to membranous vesicles secreted by cells into the extracellular environment. EV biology belongs to a rapidly developing field in biomedical sciences. EVs represent a natural mode of cell-to-cell communication, which makes them suitable for delivery of therapeutic agents, such as nucleic acids and proteins, in the body. In particular, RBCEVs feature a wide range of benefits in drug delivery as compared to extracellular vesicles derived from other cell types. In comparison to other delivery systems currently available, RBCEVs are nontoxic, low immunogenic, conveniently obtainable, and easy to use and store. Therefore, RBCEVs boast promising and exceptional advantages in overcoming various limitations of conventional therapeutics.

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Loading of Extracellular Vesicles with Chemically Stabilized Hydrophobic siRNAs for the Treatment of Disease in the Central Nervous System

Cited by 10 publications

References 28 publications

Hydrophobicity drives the systemic distribution of lipid-conjugated siRNAs via lipid transport pathways

Hydrophobicity drives the systemic distribution of lipid-conjugated siRNAs via lipid transport pathways

Therapeutic Potential of Nucleic Acids when Combined with Extracellular Vesicles

The Biology and Therapeutic Applications of Red Blood Cell Extracellular Vesicles

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