Serum-Independent Nonviral Gene Delivery to Innate and Adaptive Immune Cells Using Immunoplexes

Area of Science:

• Biotechnology and Biomedical Engineering
• Immunology and Gene Therapy

Background:

• Genetic engineering of immune cells holds promise for treating immune-mediated diseases.
• Existing methods like electroporation and viral vectors have limitations, including toxicity and efficiency issues.
• Non-viral nanoparticles offer flexibility but face challenges in stability, toxicity, and transfection efficiency, especially in serum.

Purpose of the Study:

• To develop a simple, efficient, and serum-independent non-viral gene delivery platform for immune cells.
• To engineer nanoparticles with tunable properties for precise control over cellular interactions and gene expression.
• To address limitations of current gene delivery systems, including toxicity and short-term expression.

Main Methods:

• Synthesis of cationic acetylated polyethylenimine (Ac-PEI) complexed with plasmid DNA (pDNA).
• Envelopment of Ac-PEI/pDNA complexes with poly(ethylene-alt-maleic acid) (PEMA) to form immunoplexes (IPs).
• Evaluation of IP formulation, cellular interactions, gene expression, toxicity, and serum stability in various immune cell types (macrophages, dendritic cells, T cells).

Main Results:

• Optimized IPs demonstrated potent and serum-independent transfection of innate and adaptive immune cells.
• PEMA envelopment allowed precise control over cellular interactions and targeting.
• High N/P ratios in optimally formulated IPs led to enhanced stability, reduced toxicity, high gene expression, and prolonged expression (>3 days) compared to controls.

Conclusions:

• Engineered immunoplexes (IPs) represent a simple, modular, and targetable non-viral gene delivery platform.
• This platform efficiently alters gene expression in immune cells, overcoming key limitations of existing technologies.
• The developed IPs show significant potential for clinical applications in immune cell engineering and therapy.