Investigating the expression profile of AAV and AAV retro via different CNS routes of administration
Graduation Year
2025
Document Type
Master's Thesis
Degree
Master of Science
Program
Biological Science
Partner Organization
Biomarin
Program Director
Patti Culross, MD, MPH
First Reader
Kathryn Davidson, MS, MA, PMP, RAC
Second Reader
Kerui Gong, PhD
Abstract
AAV gene therapy uses a modified non-replicating virus to deliver genetic material into cells. This method has allowed targeted and sustained expression of genetic material for therapeutic purposes. Many AAV serotypes have been identified with varying neuronal expression profiles, yet one of the main challenges with CNS gene therapy is brain delivery and controlled distribution of AAV to relevant cells. To understand the impact of delivery route on AAV expression profile, we investigated various combinations of direct CNS injections with different AAV serotypes. Retro-orbital (RO), Intracerebral ventricle (ICV), Intrathalamic, (ITh), and Intraputamen (IPu) injections were explored using 4 different AAV capsids at varying doses in adult C57BL/6 mice. Each delivery route at 3 weeks post-injection demonstrated a distinct expression profile of the capsid injected, with IPu injections producing the most extensive and robust reporter expression, particularly within forebrain areas. Furthermore, an AAV with retrograde transport capability delivered by ITh injection labelled specific layer 5 cortical circuitry, which may be advantageous. A matrix of data mapping the role of route of administration and capsid on expression profile is reported here.