Plasschaert, Lindsey W, Stutz, Cian, Otarola, Ester, Shang, Jianyu, Valdez Misiolek, Rachel, Choo-Wing, Rayman, Quigley, Catherine, Ruggeri, Lionello, Taddese, Bruck, Decock, Arnaud, Nuccio, Arthur, Watson, Rebecca, Kubek-Luck, Katie, Ibrahim Aibo, Daher, Magnifico, Maria, Buchs, Mirjam, Tan, Ge, Lin, Amy-1, Ashley, Louise, Wang, Hsu-Kun, Bardroff, Michael and Wetzel, Kristie (2025) A systemically delivered AAV-CFTR gene therapy approach for cystic fibrosis. A systemically delivered AAV-CFTR gene therapy approach for cystic fibrosis. ISSN 2692-8205

Abstract

Cystic fibrosis (CF) is the most common monogenic lung disease and results from mutations in the Cystic Fibrosis Transmembrane Conductance Regulator (CFTR). There have been over 2000 variants identified in patients that can result in loss of function of the CFTR protein leading to systemic disease and respiratory failure in adolescence. While some variants encode proteins with residual activity that can be corrected or potentiated by CFTR modulators, at least 10% of CF individuals cannot tolerate the modulators or have nonsense mutations which fail to make any protein. For all people with CF, a mutation agnostic gene replacement strategy could provide a cure for CF lung disease. Here, we propose using a systemic route of administration to deliver a functional CFTR minigene cargo with a lung tropic AAV capsid. This would serve to reach multiple organs, most importantly the lung epithelium, and would provide a functional CFTR transgene that could be expressed in any cell type with a ubiquitous promoter. To achieve this, we generated the smallest CFTR minigene tested in an AAV delivery to date. We demonstrate it is expressed and functions following transfection in cell-based assays and restores function to primary CF airway cells after viral delivery. Furthermore, we identify an AAV capsid that can transduce alveolar and airway epithelium with systemic delivery in non-human primates. These data provide tools for delivering a functional CFTR minigene that fits within the packaging capacity of an AAV and demonstration of lung transduction following systemic delivery in a large animal model. This strategy would serve to reach target airway cells while circumventing the strong mucosal barrier in CF airways and has the potential to restore CFTR function in additional CF affected organs.

Item Type:	Article
Keywords:	we generated the smallest CFTR minigene tested in an AAV delivery to date. We demonstrate its expression and function following transfection in cell-based assays and restoration of function in primary CF airway cells after viral delivery. We identify an AAV capsid that can transduce alveolar and airway epithelium with systemic delivery in non-human primates. Providing tools for delivering a functional CFTR minigene that fits within the packaging capacity of an AAV and demonstrate lung transduction with an AAV following systemic delivery in a large animal model.
Date Deposited:	08 Apr 2025 00:45
Last Modified:	08 Apr 2025 00:45
URI:	https://oak.novartis.com/id/eprint/56451