Engineering Human Circulating Monocytes/Macrophages by Systemic Deliverable Gene Editing.

Area of Science:

• Biomedical Engineering
• Cell Biology
• Gene Therapy

Background:

• Macrophage genetic engineering is crucial for treating diseases like cancer and infections.
• Plasmid DNA (pDNA) delivery into terminally differentiated macrophages is challenging due to nuclear membrane barriers.

Purpose of the Study:

• To develop an effective method for delivering plasmid DNA into macrophages for genetic engineering.
• To engineer macrophages using a novel nanoparticle (NP) system for enhanced gene delivery.

Main Methods:

• Developed core-shell nanoparticles (NPs) using a novel cationic lipid to encapsulate IL-4 pDNA (IL-4pDNA-NPs).
• Transfected human blood monocyte-derived macrophages (MDM) with IL-4pDNA-NPs.
• Assessed NP internalization, nuclear delivery, exogenous IL-4 expression, and M2 macrophage polarization in vitro and in vivo.

Main Results:

• IL-4pDNA-NPs were efficiently internalized by MDM and delivered to the nucleus within 2 hours.
• Exogenous IL-4 expression was detected within 1-2 days and persisted for up to 30 days.
• Functional IL-4 expression induced M2 macrophage polarization in vitro and in a mouse model of inflammation.

Conclusions:

• The developed NPs protect pDNA from degradation and facilitate nuclear transport, enhancing gene delivery in macrophages.
• This nanoparticle-mediated gene delivery system offers a promising strategy for macrophage-based gene therapy.
• The approach provides a new solution for gene editing in monocytes and macrophages for therapeutic applications.