Transduction of Pig Small Airway Epithelial Cells and Distal Lung Progenitor Cells by AAV4.

Oliver G Chen,Mahmoud Abou Alaiwa,Steven E Mather,Raul Villacreses,Reda E Girgis,Joseph Zabner,Christian M Brommel,Xiaopeng Li,Annie Ehler,David A Stoltz,Bradley A Hamilton

doi:10.3390/cells10051014

Oliver G Chen, Mahmoud Abou Alaiwa + Show 9 more

Open Access

https://doi.org/10.3390/cells10051014

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Journal: Cells	Publication Date: Apr 25, 2021
Citations: 4	License type: CC BY 4.0

Affiliation: Michigan State University, University of Iowa, Spectrum Health

Abstract

Cystic fibrosis (CF) is caused by genetic mutations of the CF transmembrane conductance regulator (CFTR), leading to disrupted transport of Cl− and bicarbonate and CF lung disease featuring bacterial colonization and chronic infection in conducting airways. CF pigs engineered by mutating CFTR develop lung disease that mimics human CF, and are well-suited for investigating CF lung disease therapeutics. Clinical data suggest small airways play a key role in the early pathogenesis of CF lung disease, but few preclinical studies have focused on small airways. Efficient targeted delivery of CFTR cDNA to small airway epithelium may correct the CFTR defect and prevent lung infections. Adeno-associated virus 4 (AAV4) is a natural AAV serotype and a safe vector with lower immunogenicity than other gene therapy vectors such as adenovirus. Our analysis of AAV natural serotypes using cultured primary pig airway epithelia showed that AAV4 has high tropism for airway epithelia and higher transduction efficiency for small airways compared with large airways. AAV4 mediated the delivery of CFTR, and corrected Cl− transport in cultured primary small airway epithelia from CF pigs. Moreover, AAV4 was superior to all other natural AAV serotypes in transducing ITGα6β4+ pig distal lung progenitor cells. In addition, AAV4 encoding eGFP can infect pig distal lung epithelia in vivo. This study demonstrates AAV4 tropism in small airway progenitor cells, which it efficiently transduces. AAV4 offers a novel tool for mechanistical study of the role of small airway in CF lung pathogenesis in a preclinical large animal model.

Highlights

We previously developed a method to selectively isolate the different regions of the pig airway: large airways, small airways, and distal parenchyma (Figure S1) [6]
associated virus 4 (AAV4) was detected in cells harvested from the small airways and the bronchoalveolar junction area, but rarely in the large airways (Figure 9). These results suggest that AAV4 has much higher tropism for small airways than for large airways
Our analysis of associated virus (AAV) natural serotypes using cultured primary pig airway epithelia showed that AAV4 has high tropism for pig airway epithelia, with higher transduction efficiency in small airways compared to large airways

Summary

Introduction

Cystic fibrosis (CF) is caused by genetic mutations of the CF transmembrane conductance regulator (CFTR), leading to disrupted transport of Cl− and bicarbonate [1,2]. CF lung disease manifests as chronic airway infection and inflammation, the leading causes of CF morbidity and mortality. Though CFTR potentiators and correctors have been approved to treat CF patients with certain CFTR defects [3], new therapeutic strategies are still urgently needed for CF patients not responsive to current therapy. The CF pig model, engineered with deletion or mutation of CFTR, develops pathologies that mimic human

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