A Polymeric Nanoparticle Formulation for Targeted mRNA Delivery to Fibroblasts

Area of Science:

• Biotechnology
• Nanomedicine
• Drug Delivery

Background:

• Messenger RNA (mRNA)-based therapies require safe and effective delivery systems for clinical applications.
• Polymeric nanoparticles offer versatile design possibilities for targeted therapeutic protein production.

Purpose of the Study:

• To identify and optimize polymeric nanoformulations for efficient mRNA delivery to target cells.
• To evaluate the efficacy and specificity of novel nanoformulations in vitro and in vivo.

Main Methods:

• High-throughput screening of 152 formulations using a reporter fibroblast model.
• In vitro and in vivo transfection studies comparing polymeric nanoparticles with lipid-based agents.
• Structure-activity relationship analysis of monomer combinations for optimal mRNA delivery.

Main Results:

• A specific polymeric nanoformulation demonstrated superior fibroblast transfection efficiency compared to other skin cell types.
• The identified formulation outperformed lipid-based agents in delivering Cas9 mRNA and guide RNA.
• Fibroblast tropism was linked to receptor-mediated endocytosis involving CD26 and FAP, which are overexpressed in profibrotic fibroblasts.
• Optimal mRNA delivery depended on high buffering capacity and low mRNA binding affinity for endosomal escape.

Conclusions:

• High-throughput screening is effective for rapidly designing efficient mRNA delivery systems.
• The novel polymeric nanoformulation shows significant potential for targeting fibrotic diseases.
• Understanding structure-activity relationships is crucial for developing advanced mRNA delivery platforms.