Nanochannel Electro-Injection as a Versatile Platform for Efficient RNA/DNA Programming on Dendritic Cells.

Area of Science:

• Biotechnology
• Immunotherapy
• Nanotechnology

Background:

• Dendritic cell (DC) vaccines are crucial for cancer immunotherapy.
• Efficient and safe delivery of nucleic acids into DCs is vital for DC vaccine development but remains challenging.
• Current methods often struggle to balance transfection efficiency with biosafety and preventing DC maturation.

Purpose of the Study:

• To develop and optimize a novel nanochannel electro-injection (NEI) system for efficient and safe nucleic acid delivery into dendritic cells.
• To evaluate the NEI system's performance in transfecting various nucleic acid types (DNA, mRNA, circRNA) and its impact on DC viability and maturation.
• To assess the potential of NEI for in vitro transformation of DCs for cancer vaccine applications.

Main Methods:

• Utilized a nanochannel electro-injection (NEI) system based on track-etched nanochannel membranes.
• Localized electric fields on the cell membrane using nano-sized channels for low-voltage electroporation (<30 V).
• Optimized pulse conditions for transfecting DC2.4 cells with fluorescent dyes, plasmid DNA, messenger RNA, and circular RNA (circRNA).

Main Results:

• Achieved high transfection efficiency (>70%) and biosafety (viability >85%) for various nucleic acids in DC2.4 cells using NEI.
• Successfully transfected primary mouse bone marrow DCs with circRNA with 68.3% efficiency.
• Demonstrated that NEI did not significantly affect cellular viability or induce DC maturation in primary cells.

Conclusions:

• The nanochannel electro-injection (NEI) system provides a safe and efficient platform for in vitro dendritic cell transformation.
• NEI overcomes key challenges in nucleic acid delivery for DC vaccine development.
• This technology holds significant promise for advancing the creation of effective dendritic cell-based cancer vaccines.