Cell engineering with microfluidic squeezing preserves functionality of primary immune cells in vivo

Area of Science:

• Cellular and Molecular Biology
• Immunotherapy
• Biotechnology

Background:

• Cell-based therapies hold great promise but face manufacturing challenges.
• Electroporation is a common method for genetic engineering but its cellular impact is not fully understood.

Purpose of the Study:

• To investigate the effects of electroporation on human T cell function and gene expression.
• To compare electroporation with cell squeezing as a delivery method for biomolecules into T cells.

Main Methods:

• Genome-wide transcriptomic analysis of human T cells after electroporation.
• Measurement of cytokine secretion (IL-2, IFN-γ) post-electroporation.
• In vivo assessment of T cell function, homing, and therapeutic efficacy in tumor models.
• Comparison of electroporation and cell squeezing for PD-1 editing in T cells.

Main Results:

• Electroporation caused widespread gene expression disruptions in T cells, leading to altered cytokine secretion (e.g., 648-fold IL-2 increase).
• Electroporated T cells showed functional deficiencies in vivo, failing to mount sustained antigen-specific responses.
• Cell squeezing resulted in minimal transcriptional changes and preserved T cell effector functions and therapeutic potential.
• T cells edited via cell squeezing demonstrated superior tumor-killing ability compared to those edited via electroporation.

Conclusions:

• The choice of delivery method significantly impacts cell function and therapeutic efficacy.
• Cell squeezing emerges as a superior method for engineering T cells, preserving their functional integrity.
• Further research into delivery mechanisms is crucial for advancing cell-based therapies.