Development and evaluation of novel nanoparticle formulations for therapeutic genome editing

Genetic diseases are a leading cause of death and disability in Canada and worldwide with immense economic and societal burdens. CRISPR has improved our ability to treat these diseases; however, conventional CRISPR introduces DNA breaks and delivery to affected tissues remains a challenge. Our team has engineered safer and more effective genome editors (base & prime editors) that do not introduce DNA breaks. However, the delivery of genome editors to affected cells remains a challenge. To address this challenge, in Aim 1 we will develop new nanoparticle formulations that improve the delivery of genome editors to muscle. We will start with selected, proven, nanoparticle formulations and improve their delivery to muscle through iterative rounds of optimization by varying key components and production processes and by incorporating surface peptides to target muscle. Each new formulation will first be evaluated in cell culture (in vitro) to characterize and confirm functionality. To efficiently monitor the delivery of functional genome editors in vivo, which will guide the development of improved nanoparticles, we've developed a mouse model (LumA mice) carrying a mutation in a reporter gene that emits light when repaired, allowing us to precisely monitor genome editing in living animals. In Aim 2, we will evaluate the efficacy and safety of genome editing in small and large animal disease models of a well-characterized genetic disease, Lipoprotein Lipase Deficiency. This disease is well-suited because of the severe disease features and because we can directly compare the efficacy with a previously approved viral-mediated (AAV) gene therapy. The findings from this project will be directly applicable to the development of new therapeutics for thousands of genetic diseases because the modular components can be readily tailored to treat many different diseases. Ultimately, this research will help translate new therapeutics for genetic diseases into mainstream healthcare.