Mingling life imaging and cell subpopulations expression analysis in NHP lungs targeted with exosome associated double AAV vectors

PROJECT SUMMARY
Gene therapy provides a universal therapeutic option for patients with genetic or acquired disorders that can
potentially benefit a broad patient populations. However, up to date several gene therapy clinical trials for lung-
specific diseases, while well addressing safety concerns, have failed to demonstrate clear therapeutic benefits
among the participated patients. This dismal reality is most likely attributed to the inability to achieve clinically-
relevant gene transfer efficacy in the airway epithelium, and particularly to airway basal stem cell or basal cell.
Recently, we have demonstrated the ability of AAV serotype 6 (AAV6) vectors to achieve widespread and
efficient transgene expression in the human airway epithelial cell (AEC) in in vitro and in the mouse models in
vivo. Importantly, subsets of the lung cell, such as basal cells, were targeting that provides anticipation for
possible long-term gene expression in highly renewable layer of airways. Additionally we showed in our most
recent published collaborative studies higher transduction efficiency over commonly used AAVs by
extracellular vesicles associated AAV (EV-AAV) to the mouse lungs and human AEC cultured in air-liquid
interface conditions. The large cargo of average EVs (100-200 nm) allows accumulation of multiple AAV
particles (20-25 nm), and, unlike AAV, EVs enter cells multiple by ways that do not depend on receptor
abundance. These important findings lead to the development of current proposal that combines non-invasive
in vivo imaging with post-necropsy analysis of AAV-mediated gene expression in subpopulations of airway
cells in non-human primates (NHP) model. NHP models play a critical role in feasibility and safety modeling for
the development of therapeutics. The mechanistic underpinning is expected to provide critical information for
the design of next-generation gene therapy vectors targeting lung, which can be used for wide range of
pulmonary diseases required reliable expression of therapeutic modalities across airways.

Grant Number: 1R21AI182508-01A1
NIH Institute/Center: NIH
Principal Investigator: George Aslanidi