Glycopolycation-DNA Polyplex Formulation N/P Ratio Affects Stability, Hemocompatibility, and in Vivo Biodistribution

Area of Science:

• Biomaterials Science
• Gene Therapy
• Nanotechnology

Background:

• Nonviral gene delivery methods are crucial for genome editing therapies but face limitations in biocompatibility and understanding carrier interactions in vivo.
• Developing safe and effective nonviral vectors is essential for advancing gene therapy applications for genetic diseases.

Purpose of the Study:

• To characterize novel diblock polycations, poly(methacrylamido glucopyranose- block-2-methylaminoethyl methacrylate) [P(MAG- b-MAEMt)], for nonviral gene delivery.
• To evaluate the hemocompatibility and in vivo biodistribution of P(MAG- b-MAEMt) based polyelectrolyte complexes (polyplexes) compared to control polymers.

Main Methods:

• Synthesized P(MAG- b-MAEMt) glycopolymers and control polymers [P(EG- b-MAEMt) and P(MAEMt)].
• Formulated polymers with plasmid DNA to create polyplexes at various N/P ratios.
• Assessed ex vivo hemocompatibility (red blood cell lysis, coagulation) and in vivo biodistribution of polyplexes.

Main Results:

• P(MAG- b-MAEMt) showed significantly improved hemocompatibility, with minimal red blood cell lysis compared to controls.
• Hemocompatibility was influenced by both polymer structure and N/P ratio, with N/P ratio being a stronger determinant of biodistribution.
• P(MAG- b-MAEMt) at N/P=15 demonstrated enhanced in vivo biodistribution in the liver, lungs, and spleen, indicating improved polyplex stability.

Conclusions:

• Glucose-containing diblock polycations offer a promising platform for safer nonviral gene delivery.
• Optimizing the N/P ratio is critical for tuning polyplex stability and achieving desired in vivo biodistribution for gene therapy applications.