Intradermally delivered mRNA-encapsulating extracellular vesicles for collagen-replacement therapy

Abstract

The success of messenger RNA therapeutics largely depends on the availability of delivery systems that enable the safe, effective and stable translation of genetic material into functional proteins. Here we show that extracellular vesicles (EVs) produced via cellular nanoporation from human dermal fibroblasts, and encapsulating mRNA encoding for extracellular-matrix α1 type-I collagen (COL1A1) induced the formation of collagen-protein grafts and reduced wrinkle formation in the collagen-depleted dermal tissue of mice with photoaged skin. We also show that the intradermal delivery of the mRNA-loaded EVs via a microneedle array led to the prolonged and more uniform synthesis and replacement of collagen in the dermis of the animals. The intradermal delivery of EV-based COL1A1 mRNA may make for an effective protein-replacement therapy for the treatment of photoaged skin.Recent developments in messenger RNA-modification techniques have enhanced the therapeutic efficiency of mRNA delivery and its potential for near-term clinical applications, including protein-replacement therapy and vaccination against the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus 1,2 . However, the intrinsic inability and potential immunogenicity of mRNAs require that they be encapsulated within delivery vehicles. Current mRNA-delivery modalities centre on the usage of lipid nanoparticle (LNP) carriers for encapsulation and

Small extracellular vesicles: Yields, functionalization and applications in diabetic wound management

“…With this approach, the yield and exosomal mRNA transcripts in collected engineered sEVs were both augmented markedly by 50 folds and 1000 folds, respectively 91 . Subsequently, the CNP method was utilized to create sEVs enriched with COL1A1 mRNA, serving the purpose of collagen‐replacement therapy for photoaged skin, and potentially offering insights into diabetic wound management 82 . Additionally, a brand‐new method called EXPLORs (sEVs for protein‐loading by optically reversible protein‐protein interactions) was developed to load soluble proteins inside sEVs with high efficacy 92,93 .…”

“…Additionally, the CNP method‐based electroporation was employed to fabricate COL1A1 mRNA‐enriched sEVs towards collagen‐replacement therapy in photoaged skin, shedding light on optimizing collagen deposition at diabetic wounds 82 . Hydrophobic drugs can be loaded into the lumen of sEVs using electroporation.…”

“…serving the purpose of collagen-replacement therapy for photoaged skin, and potentially offering insights into diabetic wound management. 82 Additionally, a brandnew method called EXPLORs (sEVs for protein-loading by optically reversible protein-protein interactions) was developed to load soluble proteins inside sEVs with high efficacy. 92,93 As exemplified, super-repressor IκB (srIκB, the dominant active form of IκBα) was encapsuled massively into the lumens of sEVs to alleviate sepsisrelated organ damage and inhibiting the secretion of proinflammatory cytokines.…”

Lipid Nanoparticle (LNP) Enables mRNA Delivery for Cancer Therapy

“…So far, various biologicallyderived (bio-derived) and chemically-based (chemical-based) materials have been developed to deliver mRNA, some of which have shown promising performance. [2,20] Bio-derived vehicles, including exosomes, [21][22][23] bacteria outer membrane vesicles, [24] and virus-like particles (VLPs, e.g., pseudotyped PEG10 VLPs), [25] are attractive in certain applications. Among chemical-based delivery systems, polymer-and lipid-based nanoparticles are most widely used due to their high potency and diversity.…”

Dendritic Cell‐Mimicking Nanoparticles Promote mRNA Delivery to Lymphoid Organs

“…[ 7 ] Although these formulations have feasibility for mRNA therapy, their cytotoxicity, poor biological distribution, and lack of specific targeting remain challenges for LNPs. [ 8 ] These problems may be due to LNPs requiring surface PEGylation to improve their cycle half‐life and reduce non‐specific clearance. [ 9 ] Additionally, non‐immune cells at the injection site, including muscle cells and epithelial cells, take up the majority of the injected LNPs but do not participate in the antigen presentation that occurs in the draining lymph nodes.…”