Cell-specific mRNA delivery via nanobody-functionalized lipid nanoparticles

Area of Science:

• Biotechnology
• Nanomedicine
• Molecular Biology

Background:

• Messenger RNA–lipid nanoparticle (mRNA-LNP) formulations are a powerful platform for various therapies.
• A key limitation is the absence of targeted delivery, hindering efficacy and safety.
• Cell-specific delivery is crucial for advancing mRNA therapeutic applications.

Purpose of the Study:

• To engineer a novel mRNA-LNP platform with targeted delivery capabilities.
• To functionalize lipid nanoparticles with nanobodies targeting aminopeptidase N (APN) on gut epithelial cells.
• To demonstrate enhanced cellular uptake and delivery of mRNA payloads via targeted functionalization.

Main Methods:

• Lipid nanoparticles were functionalized with nanobodies (VHH) specific to aminopeptidase N (APN).
• Nanobodies were engineered in E. coli to incorporate azido-phenylalanine for click chemistry conjugation.
• Targeted conjugation onto lipid nanoparticles was achieved using SPAAC and IEDDA click chemistry.
• The efficacy of targeted delivery was assessed in porcine intestinal organoids and in vivo models.

Main Results:

• APN-targeted mRNA-LNPs demonstrated selective uptake by APN-expressing cells.
• Functionalized LNPs enhanced mRNA delivery to target cells.
• Nanobody-mediated targeting promoted transcytosis across the gut epithelial barrier.
• The platform showed successful delivery in both in vitro and in vivo models.

Conclusions:

• The developed nanobody-functionalized mRNA-LNP platform enables targeted delivery of mRNA payloads.
• This programmable system enhances cellular uptake and delivery across the gut barrier.
• The approach offers a versatile solution for cell-specific mRNA delivery, adaptable across species.
• This technology holds significant potential for advancing mRNA-based vaccines and therapeutics.