A hybridized mechano-electroporation technique for efficient immune cell engineering

Dorsa Morshedi Rad; William P Hansen; Sareh Zhand; Charles Cranfield; Majid Ebrahimi Warkiani

doi:10.1016/j.jare.2023.11.009

A hybridized mechano-electroporation technique for efficient immune cell engineering

J Adv Res. 2023 Nov 11:S2090-1232(23)00346-6. doi: 10.1016/j.jare.2023.11.009. Online ahead of print.

Authors

Dorsa Morshedi Rad¹, William P Hansen², Sareh Zhand¹, Charles Cranfield², Majid Ebrahimi Warkiani³

Affiliations

¹ School of Biomedical Engineering, University of Technology Sydney, Sydney, NSW 2007, Australia.
² School of Life Sciences, University of Technology Sydney, Sydney, NSW 2007, Australia.
³ School of Biomedical Engineering, University of Technology Sydney, Sydney, NSW 2007, Australia; Institute for Biomedical Materials and Devices (IBMD), Faculty of Science, University of Technology Sydney, Sydney, NSW 2007, Australia. Electronic address: majid.warkiani@uts.edu.au.

PMID: 37956863
DOI: 10.1016/j.jare.2023.11.009

Abstract

Immune cell engineering, which involves genetic modification of T cells, natural killer cells, and macrophages, is shifting the paradigm in immunotherapy for treating hematologic malignancies. These modified cells can be viewed as living drugs and offer advantages, including dynamic functionality, active local trafficking, and boosting the immune system while recognizing and eliminating malignant cells. Among the current technologies employed for the modification of immune cell functions, electroporation stands as a predominant approach, but it suffers from heterogeneity arising from the treatment of a bulk population of immune cells during the manufacturing procedures. To address this challenge of the field, here we present a hybrid approach to induce consecutive gentle mechanical and electric shocks. This approach enhances the treatment homogeneity and improves outcomes in difficult-to-load immune cells. The hybrid approach aims to enhance the treatment homogeneity by passing individual immune cells through a microengineered filter membrane with micropores smaller than the cell diameter. This facilitates the creation of transient pores in the cell membrane, followed by efficient delivery of biomolecules through the complementary use of a gentle electric shock. Using this hybrid mechano-electroporation (HMEP) system, we could successfully deliver fluorescein isothiocyanate (FITC) dextran molecules from the smallest (4 kDa) to the largest (2000 kDa) size and EGFP expressing plasmid DNA into different immune cell types. We also provide insight into the delivery performance of the HMEP system in comparison with the benchtop electroporation since both methods hinge on membrane disruption as their permeabilization mechanism. Immune cells treated with the HMEP protocol demonstrated higher delivery efficiencies while maintaining cell viability compared to those experiencing conventional electroporation. Therefore, membrane-based mechanoporation can be a cost-effective and efficient approach to pre-treat the hard-to-deliver immune cells before electroporation, elevating the treatment homogeneity and delivery of exogenous cargoes to a higher level.

Keywords: Cancer immunotherapy; Hybrid mechano-electroporation protocol; Immune cell engineering; Membrane disruption.