SENOLYTIC CRISPR CAR T CELLS PRODUCED BY CRISPR-CAS9 GENOME EDITING

Smart overview of the Invention

Utilizing CRISPR-Cas9 genome editing, methods have been developed to create chimeric antigen receptors (CARs) on T cells. These CAR T cells specifically target the urokinase Plasminogen Activator Receptor (uPAR) found on senescent cells. This approach aims to improve therapies for conditions such as neurodegenerative diseases, stroke, and trauma, particularly in elderly patients.

Challenges of Senescent Cells

Cellular senescence is a response to stress that results in cell cycle arrest, contributing to tissue dysfunction. Senescent cells secrete factors that can attract immune cells; however, their clearance is often hindered in aged tissues. Current treatments, known as senolytic therapies, show promise but require enhanced strategies for effectively eliminating these cells.

Innovative CAR T Cell Design

The new approach incorporates a DNA homology-directed repair (HDR) template designed to insert a CAR gene into T cell genomes. This process enables the generation of CAR T cells with specific targeting capabilities against uPAR+ senescent cells. The template includes various components that facilitate the expression and functionality of the CAR, enhancing the therapeutic potential of these engineered T cells.

Ex Vivo Production Method

A detailed method outlines how to produce these CAR T cells without using viruses. By introducing a Cas9 ribonucleoprotein complex along with the HDR template into unmodified T cells, researchers can achieve precise genomic integration of the CAR transgene. The resulting CAR T cells are cultured in a xeno-free medium, ensuring safety and efficacy in therapeutic applications.

Applications in Neurodegenerative Diseases

The engineered CAR T cell therapies are particularly relevant for treating neurodegenerative conditions like Alzheimer's Disease and Parkinson's Disease, where senescent cell accumulation is prevalent. By targeting and eliminating these cells, the therapies aim to reduce inflammation and restore tissue function, representing a significant advancement in treating age-related diseases.