Bottom-up manufacturing of artificial anti-tumor T cells

Introduction

T cells play a central role in anti-tumor immune protection. While their ability to target and eliminate emerging tumor cells is increasingly recognized, fully-established tumors can efficiently evade T cell response.

Background

Significant efforts spanning several decades of research have been made to develop T cell-based therapies manufactured from donor-derived T cells. The use of tumor-directed T cells engineered to express chimeric antigen receptors (CARs) represents, to date, one of the most successful applications for the treatment of chemoresistant cancers.

Challenges

However, several major drawbacks are still hindering the full potential of T cell-based therapies, including:

1. Economic factors
2. Suboptimal functioning
3. Life-threatening side effects

Objective

To address these issues, we aim to generate Artificial T cells (ArTCell) that will mimic the anti-tumor function of a T cell-based therapy but in a safer, more efficient, and less expensive product.

Design and Features

ArTCells will incorporate two key features of activated T cells into Giant Unilamellar Vesicles (GUVs):

• The specificity of tumor cell recognition
• The cytotoxic activity achieved through death ligands and cytolytic proteins

Methodology

Functionalization of the GUVs will be confirmed by:

• Immunofluorescent labeling of membrane proteins (i.e., TRAIL, LFA-1, and CAR)
• Detection with flow cytometry

The morphology of ArTCell will be monitored via:

• Cryo-EM
• SEM
• Confocal microscopy

Validation

The ability of ArTCell to target and kill tumor cells will be thoroughly validated in vitro by a combination of functional and high-resolution live imaging assays, as well as in vivo with two cell line- and patient-xenograft mouse models.

Conclusion

The ArTCell could allow us to circumvent many of the current technological limitations that hinder a more widespread applicability of cell-based therapies, without being subject to tumor-mediated inactivation.