Abstract
Application of in vitro transcribed (IVT) messenger ribonucleic acid (mRNA) is an increasingly popular strategy to transiently produce proteins as therapeutics in a tissue or organ of choice. Here, we focused on the skin and aimed to test if whole human skin tissue explant technology can be used to evaluate the expression efficacy of different IVT Interferon alpha (IFN-α) mRNA constructs in situ, after biolistic delivery. Skin explants were viable and intact for at least five days based on histologic analysis and TUNEL staining. Using GFP reporter mRNA formulations, we found mostly epidermal expression after biolistic delivery. Two out of five sequence-optimized IFN-α mRNA variants resulted in significantly improved IFN-α protein expression in human skin compared to native IFN-α mRNA transfection. IFN-α secretion analysis of the surrounding culture media confirmed these results. We provide a proof-of-concept that IFN-α mRNA delivery into intact human full thickness skin explants can be utilized to test mRNA sequence modifications ex vivo. This approach could be used to develop novel mRNA-based treatments of common epidermal skin conditions including non-melanoma skin cancer, where IFN-α protein therapy has previously shown a strong therapeutic effect.
Highlights
Recent developments in nucleic acid-based therapeutics, especially ribonucleic acid (RNA)-based therapies, have shown significant potential for a wide range of human diseases
Clinical applicability of in vitro transcribed (IVT) messenger ribonucleic acid (mRNA) for treatments other than vaccination still faces several challenges including, most notably, limited availability of custom-tailored delivery systems and lack of well-tolerated formulations reproducibly allowing for efficient mRNA expression
We showed that IVT IFN-α mRNA can be successfully delivered to human skin by biolistic application
Summary
Recent developments in nucleic acid-based therapeutics, especially ribonucleic acid (RNA)-based therapies, have shown significant potential for a wide range of human diseases (reviewed in[1]). Delivery of nucleic acids to the skin is advantageous because it is able to bypass many of these challenges associated with systemic administration, including simplifying application and increasing local bioavailabilty (reviewed in3), making it an ideal target for the development of RNA-based therapeutics. Surgical removal of NMSC lesions remains the gold-standard for treatment, it is not able to address the field-cancerization that is commonly associated with the condition, where a large surface area can be affected. Its broad implementation in the clinic for NMSC therapy has been hindered for several reasons including the need of daily applications over 10–14 days as well as high costs of IFN-α
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