Identification of AAV serotypes for lung gene therapy in human embryonic stem cell-derived lung organoids
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(2020) 11:448
SHORT REPORT
Open Access
Identification of AAV serotypes for lung gene therapy in human embryonic stem cell-derived lung organoids Helena Meyer-Berg1 , Lucia Zhou Yang2 , María Pilar de Lucas3 , Alberto Zambrano2 , Stephen C. Hyde1 and Deborah R. Gill1*
Abstract Gene therapy is being investigated for a range of serious lung diseases, such as cystic fibrosis and emphysema. Recombinant adeno-associated virus (rAAV) is a well-established, safe, viral vector for gene delivery with multiple naturally occurring and artificial serotypes available displaying alternate cell, tissue, and species-specific tropisms. Efficient AAV serotypes for the transduction of the conducting airways have been identified for several species; however, efficient serotypes for human lung parenchyma have not yet been identified. Here, we screened the ability of multiple AAV serotypes to transduce lung bud organoids (LBOs)—a model of human lung parenchyma generated from human embryonic stem cells. Microinjection of LBOs allowed us to model transduction from the luminal surface, similar to dosing via vector inhalation. We identified the naturally occurring rAAV2 and rAAV6 serotypes, along with synthetic rAAV6 variants, as having tropism for the human lung parenchyma. Positive staining of LBOs for surfactant proteins B and C confirmed distal lung identity and suggested the suitability of these vectors for the transduction of alveolar type II cells. Our findings establish LBOs as a new model for pulmonary gene therapy and stress the relevance of LBOs as a viral infection model of the lung parenchyma as relevant in SARSCoV-2 research. Keywords: human embryonic stem cells, stem cell-based tissue model, gene therapy, rAAV, AAV capsids, AAV serotypes, lung organoids, viral infection model, alveolar type II cells
Background Recombinant adeno-associated virus (rAAV) is a wellestablished vector for gene delivery, currently in use clinically for gene therapy, with multiple, naturally occurring serotypes and artificial variants facilitating species-specific cell and tissue tropisms [1]. Engineering of new AAV capsids has been the focus of extensive research, but capsids selected in animal models and cancer cell lines often translate poorly to large animal models and humans. Clinical trials of gene therapy for cystic * Correspondence: [email protected] 1 Gene Medicine Research Group, Nuffield Division of Clinical Laboratory Sciences, Radcliffe Department of Medicine, University of Oxford, Oxford, UK Full list of author information is available at the end of the article
fibrosis lung disease using AAV serotype 2 failed to show efficacy [2], and of the many potential reasons for this, an important factor is the lack of serotype screening for airway tropism resulting in poor translation to the human airways [3]. The identification of AAV serotypes for gene delivery to the human lung has focused mainly on the transduction of the human airway epithelium [4, 5]. The lung parenchyma, however, is the target for treating genetic diseases
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