Optimizations of In Vitro Mucus and Cell Culture Models to Better Predict In Vivo Gene Transfer in Pathological Lung Respiratory Airways: Cystic Fibrosis as an Example

Abstract

The respiratory epithelium can be affected by many diseases that could be treated using aerosol gene therapy. Among these, cystic fibrosis (CF) is a lethal inherited disease characterized by airways complications, which determine the life expectancy and the effectiveness of aerosolized treatments. Beside evaluations performed under in vivo settings, cell culture models mimicking in vivo pathophysiological conditions can provide complementary insights into the potential of gene transfer strategies. Such models must consider multiple parameters, following the rationale that proper gene transfer evaluations depend on whether they are performed under experimental conditions close to pathophysiological settings. In addition, the mucus layer, which covers the epithelial cells, constitutes a physical barrier for gene delivery, especially in diseases such as CF. Artificial mucus models featuring physical and biological properties similar to CF mucus allow determining the ability of gene transfer systems to effectively reach the underlying epithelium. In this review, we describe mucus and cellular models relevant for CF aerosol gene therapy, with a particular emphasis on mucus rheology. We strongly believe that combining multiple pathophysiological features in single complex cell culture models could help bridge the gaps between in vitro and in vivo settings, as well as viral and non-viral gene delivery strategies.

Muscarinic receptor antagonists and airway inflammation: A systematic review on pharmacological models

“…Immortalized cell lines originating from human respiratory tract represent a simple and valuable tool to investigate airway cellular responses [ 77 ], however due to the transformation process and homogenous clonality [ 78 ], they lack all the underlying genetic or epigenetic hallmarks characterizing the original tissue [ 79 ].…”

Apparent Yield Stress of Sputum as a Relevant Biomarker in Cystic Fibrosis

“…Interestingly, the apparent yield stress, rather than the linear viscoelastic moduli G and G and even the Newtonian viscosity, turned out to be the most relevant biomarker for the development and the monitoring of mucolytic agents acting on the DNA/actin network. This could also be used as a key parameter to study the efficiency of new pharmacological therapies such as Trikafta ® or prior to gene therapy delivery, as well as in the development of in vitro mucus models for the screening of new drugs or the improvement of their formulations [38,39].…”

Aerosol-Mediated Non-Viral Lung Gene Therapy: The Potential of Aminoglycoside-Based Cationic Liposomes

“…Altogether, the results obtained from a dose escalation study (Figure S7) suggest that, although they are more sensitive than GL67A, CholP-based lipoplexes may retain some transfection efficiency under CF relevant conditions. This also suggests the requirement to further optimize CholP formulation, especially as regards its content in DP5K, and to investigate more complex cell culture models [70,71]. (ii) Considering bacterial infections that occur in the airways of CF patients, additional antibacterial properties of cationic lipids [18] or polymers [72] might be useful for CF lung gene therapy applications.…”