PEN photoporation for the genetic engineering of therapeutic mesenchymal stromal cells and T cells

Introduction

Adoptive cell therapy has emerged as a promising strategy to treat cancer. It relies on patient-derived cells, such as T cells and mesenchymal stromal cells (MSCs), which are genetically engineered to become better equipped to fight cancer cells.

Challenges with Traditional Methods

While ex vivo genetic modification of T cells and MSCs has traditionally been performed with viral vectors, they come with concerns about:

• Safety
• Sustainable production
• High development costs

Alternative Transfection Technologies

Electroporation is a non-viral alternative transfection technology, but it can lead to significant gene expression changes, phenotypic alterations, and decreased therapeutic potency.

Introduction of PEN Photoporation

Recently, photoporation with electrospun photothermal nanofibers (PEN photoporation) was demonstrated to provide a safer alternative with minimal impact on the cell’s functionality and phenotype.

How PEN Photoporation Works

The technology makes use of photothermal nanofibers which, upon stimulation with laser light, can transiently permeabilize cells to allow gene-modifying effector molecules to enter the cells.

Project Objectives

Having been thoroughly demonstrated and validated in a research setting (TRL4), this project aims to bring the PEN photoporation technology to TLR6 by:

1. Developing hard- and software for automated high-throughput transfections of T cells (>1B cells/h) and MSCs (>10M cells/h).
2. Extensively testing and validating the technology in the cGMP compliant laboratories of the project partners for the genetic engineering of T cells and MSCs.
3. Preparing for commercialization and market deployment.

Expected Outcomes

By the end of the project, a fully automated and validated high-throughput prototype system will be available for:

• Installation at centralized cell production facilities
• Integration in point-of-care cell manufacturing equipment

Alignment with Micro-Nano-Bio Challenge

This project aligns with the Micro-Nano-Bio challenge as it combines nanotechnology with microfluidics to enhance genetically engineered cell therapy products.