Development of T cell Targeting CRISPR Knock-in Lipid Nanoparticles (CAR T LNPs) for In Vivo CAR T Cell Generation

• Applicant: Dr. Michael Launspach
• Subject Area: Hematology, Oncology ; Pediatric and Adolescent Medicine
• Term: since 2025
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 571675766

This project aims to develop a new, non-viral technology that enables the generation of cancer-fighting immune cells – known as CAR T cells – through a simple injection. CAR T cells are a promising form of immunotherapy where a patient’s own T cells are genetically modified to recognize and destroy cancer cells. Currently, making CAR T cells is a complicated process: T cells are removed from the patient, modified in the lab, and reinfused. This “ex vivo” approach is expensive, time-consuming, and only available at select centers. Our goal is to replace this complex process with a single, injectable therapy. To do this, we are developing tiny delivery systems called lipid nanoparticles (LNPs), similar to those used in mRNA COVID-19 vaccines. These LNPs carry the tools needed to rewrite the DNA of T cells inside the body using CRISPR–Cas9 gene editing technology. Specifically, the LNPs deliver: Cas9 – the “molecular scissors,” guide RNA – which directs Cas9 to the correct spot in the genome, and a DNA repair template (HDRT) – which inserts the genetic code for the cancer-targeting receptor (CAR). The aim is to insert this CAR precisely into a specific location in the T cell genome (the TRAC locus), allowing long-lasting and tightly controlled CAR expression. To improve the efficiency and safety of this process, we use innovative features: DNA packaged in special nanostructures (“DNA origami”) for better stability and delivery into the cell nucleus, biodegradable polymers with nuclear targeting signals, and chemical modifications that help the nanoparticles reach T cells specifically, while avoiding detection by the immune system. The LNP surface is engineered with targeting molecules (called scFvs) that bind only to T cells, and “cloaking” peptides to minimize unwanted immune responses. These innovations come from world-leading labs at UC Berkeley and UCSF, including optimized lipids and delivery chemistries. The gene editing machinery delivered by the LNPs is active only for a short time, reducing the risk of long-term side effects. First, we will test LNP versions in human T cells and immune cells in the lab to identify the most effective and safest combinations. The best candidates will then be tested in mice with human immune cells to assess whether they can safely generate CAR T cells inside the body and eliminate leukemia cells. The project builds on extensive prior work by the applicant and a strong collaboration network. If successful, this research could transform cancer treatment by allowing personalized cell therapies to be delivered as a single injection – without hospitalization or complex lab procedures. It lays the foundation for safe, scalable, and globally accessible cell-based immunotherapies of the future.

• DFG Programme: WBP Fellowship
• International Connection: USA
• Host: Professor Ross Wilson, Ph.D.