NOn-VIral gene modified STEM cell therapy

Introduction

Despite the increasing awareness that cell and gene-therapy approaches have tremendous biomedical potential, their broad clinical application has been challenging due to prolonged and expensive production times and the emergence of severe immune- and gene-delivery dependent side effects.

Objective

In this proposal, we aim to establish a streamlined and high-throughput protocol for iPSC-based cell therapy by combining a novel technological platform for gene delivery with a breakthrough biological concept that will permit the manufacture of functional, gene-corrected blood-forming stem cells and CAR T cells.

Methodology

To achieve this, we will:

1. Use and optimize photoporation as a non-viral gene delivery method for CRISPR-mediated and site-specific gene editing.
2. Obtain controlled CAR expression and perform gene correction in iPSCs.
3. Generate CAR T cells and blood-forming stem cells from these gene-modified iPSCs by selectively targeting a signaling pathway that we established to be critical in human blood cell development and particularly T cell development.

Validation and Optimization

Following functional validation of the generated cell products, we will optimize the current protocols to increase the potential for clinical implementation.

High-Throughput Platform

We will establish a high-throughput photoporation platform to generate a large number of CAR-expressing iPSC lines from different ages, sexes, and ethnicities to demonstrate the population-wide implementation potential of our approach.

Impact

This will allow us to generate a bank of well-characterized, HLA-defined CAR-expressing iPSCs that can be used as off-the-shelf cell therapy products. This approach will significantly advance the currently implemented adaptive CAR T cell approaches by:

• Reducing production costs and time.
• Selectively targeting the CAR into a well-controlled location, which will prevent variability.
• Facilitating the production and evaluation of novel CARs for other cancer entities such as solid tumors.