Multilayered Cultures of NSCLC cells grown at the Air-Liquid Interface allow the efficacy testing of inhaled anti-cancer drugs

Dania Movia,Yuri Volkov,Despina Bazou,Adriele Prina-Mello

doi:10.1038/s41598-018-31332-6

Dania Movia, Yuri Volkov + Show 2 more

Open Access

https://doi.org/10.1038/s41598-018-31332-6

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Abstract

Evidence supports the advantages of inhalation over other drug-administration routes in the treatment of lung diseases, including cancer. Although data obtained from animal models and conventional in vitro cultures are informative, testing the efficacy of inhaled chemotherapeutic agents requires human-relevant preclinical tools. Such tools are currently unavailable. Here, we developed and characterized in vitro models for the efficacy testing of inhaled chemotherapeutic agents against non-small-cell lung cancer (NSCLC). These models recapitulated key elements of both the lung epithelium and the tumour tissue, namely the direct contact with the gas phase and the three-dimensional (3D) architecture. Our in vitro models were formed by growing, for the first time, human adenocarcinoma (A549) cells as multilayered mono-cultures at the Air-Liquid Interface (ALI). The in vitro models were tested for their response to four benchmarking chemotherapeutics, currently in use in clinics, demonstrating an increased resistance to these drugs as compared to sub-confluent monolayered 2D cell cultures. Chemoresistance was comparable to that detected in 3D hypoxic tumour spheroids. Being cultured in ALI conditions, the multilayered monocultures demonstrated to be compatible with testing drugs administered as a liquid aerosol by a clinical nebulizer, offering an advantage over 3D tumour spheroids. In conclusion, we demonstrated that our in vitro models provide new human-relevant tools allowing for the efficacy screening of inhaled anti-cancer drugs.

Highlights

Evidence supports the potential advantages of inhalation over intravenous/oral drug administration routes in the treatment of respiratory diseases[3] such as lung cancer[4]
In Air-Liquid Interface (ALI) in vitro models, drug deposition and dissolution occur in a small volume of cell lining fluid and mimic closely the delivery of liquid drug aerosol on the lung surface of human patients
Our study aimed at creating novel in vitro models that incorporated both the ALI culturing conditions and the 3D architecture of the tumour tissue, which is generally mimicked in in vitro cancer research experiments through the adoption of 3D tumour spheroids

Summary

Introduction

Evidence supports the potential advantages of inhalation over intravenous/oral drug administration routes in the treatment of respiratory diseases[3] such as lung cancer[4]. Despite suffering from poor lung deposition[5], which may cause inadequate patient compliance, inhalation allows for the administration of lower drug doses than the systemic delivery. The state-of-the-art in vivo and in vitro lung tumour models presented above highlight the compelling need for the development of preclinical models ensuring that the data generated bears a higher relevance to humans than animal studies or conventional in vitro testing based on 2D cultures. This will minimize the technical and physiological gap impacting the translation of inhaled anticancer drugs into clinical practice[16]. These in vitro platforms are better models of the biological and biochemical characteristics of human tissues than conventional 2D cell cultures

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