Identification of transfected cell types following non‐viral gene transfer to the murine lung

Abstract

Direct visualisation of GFP expression was used to detect transfected cell types in the mouse lung. In contrast with observations made using beta-galactosidase as a reporter, gene expression from several non-viral GTAs was readily demonstrated and no false GFP-positive cells were ever detected in untreated lung tissues. Lung delivery of different GTAs resulted in GFP expression in different cell types, confirming the importance of identification of transfected cells when screening and selecting GTAs for disease targets.

Efficient in vivo transfection and safety profile of a CpG-free and codon optimized luciferase plasmid using a cationic lipophosphoramidate in a multiple intravenous administration procedure

“…The route of administration can lead to the transfection of different cell types. Indeed, the nebulization of non-viral gene delivery systems often led to the transfection of epithelial cells such as ciliated epithelial cells [36,37] or alveolar type 1 pneumocytes [37,38] which are both slowly dividing or terminally differentiated [6]. Although we have shown that IV injection of lipoplexes could also conduct to a gene transfer in type 1 and type 2 pneumocytes [4], the systemic delivery procedure often leads to a transgene expression in various cell populations including endothelial cells [39,40], macrophages and others that may have higher rates of turnover.…”

Synthetic vectors for gene delivery: An overview of their evolution depending on routes of administration.

“…Low MW PEI does not induce inhibition of luciferase expression compared to PEI polymers with a high MW which can been explained by the cytotoxicity associated with high MW PEI [54]. Significant gene expression in lungs was detected in rodents [55] and in sheep [56], specifically in ciliated epithelial cells of the whole murine respiratory system [39], and alveolar type I pneumocytes [39,57]. In parallel, the 22 kDa linear PEI used for systemic administration can effectively transfect several organs in neonatal mice [58].…”

“…In numerous preclinical studies, GL67A was delivered by nasal instillation, and transgene expression was mainly localized in type I pneumocytes [39]. Administered in the same way, naked pDNA and pDNA-PEI resulted in the expression of the transgene in ciliated and Clara cells of the conducting airway [39]. BGTC is efficient for gene delivery into primary cells for human lung epithelium and mice airway cells after intratracheal administration of lipoplexes [40].…”

“…The GL67A formulation, associated with DOPE and a steric stabilizer (DMPE-PEG 5000 ) is used for running aerosolization multidose clinical assays for CF in United Kingdom [38] [http://www.cfgenetherapy.org.uk]. In numerous preclinical studies, GL67A was delivered by nasal instillation, and transgene expression was mainly localized in type I pneumocytes [39]. Administered in the same way, naked pDNA and pDNA-PEI resulted in the expression of the transgene in ciliated and Clara cells of the conducting airway [39].…”

Nanocomplexes for gene therapy of respiratory diseases: Targeting and overcoming the mucus barrier

“…Administration of the PEI-DNA complexes through the airways resulted in transfection of various cell types such as epithelial cells, macrophages, and endothelial cells [26,27]. Due to the lack of specific mechanisms involved in PEI polyplexes uptake, i.e., occurring by adsorptive endocytosis, PEI has been modified (see below Section 3.1.1) in order to increase its targeting to the airways.…”