Improving CAR-T cell therapies through AAV-mediated genetic engineering

Introduction

T cells expressing chimeric antigen receptors (CAR) have transformed cell therapies against some hematological cancers. As a postdoc, I evolved a synthetic adeno-associated virus (AAV) with tropism against murine T cells, providing a unique tool to study gene-targeted T cells in immunocompetent cancer models.

Proposal Overview

In this proposal, capitalizing on my breakthrough, I will develop novel strategies to generate gene-targeted CAR-T cells in vivo. This will involve:

1. Optimizing AAV delivery in immunocompetent mouse models.
2. Combining these methods with technologies for Cas9 delivery for T cell-specific gene editing.

The ultimate goal of my proposal is to develop methods that can be translated to clinical trials in humans.

Humanized Mouse Model

To achieve this, I will establish a humanized mouse model that allows for targeting of human T cells in vivo. Key findings from this research project will be translated for proof-of-concept experiments. As the first-ever study of gene-targeted T cells in vivo, this groundbreaking research will provide in-depth profiling of in vivo engineered CAR-T cells and their therapeutic potential.

This study is a necessary first step towards accessible and affordable in vivo generated CAR-T cell therapies in humans.

Extension to Solid Tumors

Furthermore, to extend the use of CAR-T cells against solid tumors, I have developed an AAV-based platform to perform pooled knock-in T cell screens in immunocompetent solid tumor mouse models.

For this research proposal, I have designed a library of synthetic costimulatory receptors to be expressed with a CAR at the Trac locus to improve T cell fitness and persistence.

Advanced T Cell Engineering

By combining advanced T cell engineering with analysis on a single-cell level, these pioneering experiments will answer crucial questions for T cell therapies and tumor biology.

Collaborative Environment

To succeed with my ambitious and unconventional proposal, I plan to join the Department of Medicine, Huddinge, at the Karolinska Institute. This will involve building a collaborative team in an excellent translational research environment.