Editing Hematopoietic Stem Cells for HIV Treatment

Abstract

Advances in genome editing technologies have opened the door to one-time treatments for previously incurable, chronic diseases. This has accelerated interest in delivering gene therapies and other biologic drugs with durable therapeutic effects. Hematopoietic stem and progenitor cells (HSPC) are a promising target for gene therapy as they provide a steady supply of cells throughout the body over long periods of time. In this dissertation, I explore the use of CRISPR/Cas systems in HSPC to achieve site-specific gene knock-in and to produce antibodies against HIV in vivo. The editing approach described here has been optimized to target the endogenous locus for antibody expression in primates and does not rely on viral vectors. Following transplantation in an animal model, edited HSPC are capable of self-renewal and development into mature hematopoietic cell types. Anti-HIV antibodies are detected in these animals, demonstrating productive knock-in of our gene encoding DNA template. These findings are a proof-of-concept for non-viral HSPC gene editing as a platform for durable biologics production in the treatment of chronic diseases.