A human surfactant B deficiency air-liquid interface cell culture model suitable for gene therapy applications

Altar M Munis,Stephen C Hyde,Deborah R Gill

doi:10.1016/j.omtm.2020.11.013

Altar M Munis, Stephen C Hyde + Show 1 more

Open Access

https://doi.org/10.1016/j.omtm.2020.11.013

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Abstract

Surfactant protein B (SPB) deficiency is a severe monogenic interstitial lung disorder that leads to loss of life in infants as a result of alveolar collapse and respiratory distress syndrome. The development and assessment of curative therapies for the deficiency are limited by the general lack of well-characterized and physiologically relevant in vitro models of human lung parenchyma. Here, we describe a new human surfactant air-liquid interface (SALI) culture model based on H441 cells, which successfully recapitulates the key characteristics of human alveolar cells in primary culture as evidenced by RNA and protein expression of alveolar cell markers. SALI cultures were able to develop stratified cellular layers with functional barrier properties that are stable for at least 28 days after air-lift. A SFTPB knockout model of SPB deficiency was generated via gene editing of SALI cultures. The SFTPB-edited SALI cultures lost expression of SPB completely and showed weaker functional barrier properties. We were able to correct this phenotype via delivery of a lentiviral vector pseudotyped with Sendai virus glycoproteins F/HN expressing SPB. We believe that SALI cultures can serve as an important in vitro research tool to study human alveolar epithelium, especially for the development of advanced therapy medicinal products targeting monogenic disorders.

Highlights

In the lungs, surfactant protein B (SPB) is produced by alveolar type II (ATII) and club cells.[1]
H441 cells demonstrate ATII cell characteristics A549 and H441 lung adenocarcinoma cells have been widely used as cell culture models for the lung parenchyma in drug discovery and epithelial transport studies,[23,24,25,26,27] and we investigated their potential to serve as a model for surfactant deficiencies
air-liquid interface (ALI) cultures were established from A549 cells, H441 cells, and co-culture of both lines grown in either “base” or “polarization” media.[28]

Summary

Introduction

Surfactant protein B (SPB) is produced by alveolar type II (ATII) and club cells.[1]. We propose a curative gene therapy approach for the treatment of SPB using our simian immunodeficiency virus (SIV)-based lentiviral vector (LV) pseudotyped with Sendai virus glycoproteins F and HN (rSIV.F/HN).[9] non-viral gene therapy approaches have achieved correction of the defect in mouse models,[10,11] the therapeutic effect was short-lived and inefficient. Using rSIV.F/HN, which is optimized for pulmonary gene transfer, curative therapy can be achieved lasting the patient’s lifetime or until a suitable donor organ becomes available

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