Synthesis, characterization, and evaluation of low molecular weight poly(β-amino ester) nanocarriers for enhanced T cell transfection and gene delivery in cancer immunotherapy

Abstract

Cancer immunotherapy represents a revolutionary approach in cancer treatment by leveraging the body's immune system to target and eliminate cancer cells. An emerging strategy within this field is gene delivery, which can enhance the efficacy of immune cells. Nanocarrier-based gene delivery methods have gained prominence due to their ability to protect and transport genetic material into cells efficiently. Polymeric nanocarriers, in particular, offer significant advantages, such as customizable physical and chemical properties, biocompatibility, and the potential for targeted delivery. Among polymeric nanocarriers, poly(β-amino ester) (PBAE) polymers are notable for their biodegradability, low cytotoxicity, and high gene transfection efficiency. This study investigates the synthesis and characterization of low molecular weight PBAE nanocarriers, assessing their potential in gene delivery applications for Jurkat and primary T cells—both crucial in cancer immunotherapy. Our research involved synthesizing PBAE polymer and creating nanocarriers at various DNA-to-polymer ratios. We characterized these nanocarriers in terms of size, zeta potential, and encapsulation efficiency. Confocal microscopy and flow cytometry were utilized to evaluate cellular uptake and transfection efficiency. The results demonstrated appropriate transfection efficiency and significant gene expression in both hard-to-transfect cell types (jurkat up to 37% and primary T cell 5%), with optimized DNA-to-polymer ratios showing minimal cytotoxicity. This study highlights the potential of PBAE nanocarriers in enhancing gene delivery for cancer immunotherapy. By effectively transfecting T cells, these nanocarriers could improve the therapeutic outcomes of immunotherapy, offering a promising pathway for developing more effective cancer treatments.