Airway epithelial cell model for efficacy studies with inhaled anti-inflammatory drugs


Figure 1: Calu-3 cells were stimulated with 8 mg/ml LPS in 300 µl apical medium for 24h on das 11 in culture. Prior to LPS treatment, the cells were pre-incubated for 30 min with the MAPK inhibitor SB203580 (A) or with the IKK-2 inhibitor TPCA-1 (B, C). The secretion of IL-6 (A, B) or IL-8 (C) was quantified by ELISA.

The Calu-3 cell line is a well characterized model of the airway epithelial barrier and has been frequently used to study the uptake and transepithelial transport of drugs. To resemble a chronic inflammatory state, as observed in many pulmonary diseases, such as Asthma and COPD, the cells are treated with LPS. The efficacy of anti-inflammatory drugs is then measured by means of functionality, i.e. reduction of proinflammatory cytokine secretion. Further endpoints addressing toxicity or specific cellular effects (e.g. mucus production) can be included in the model.

Species

human

Fields of application

  • Screening of new drug candidates for anti-inflammatory efficacy
  • Absorption studies
  • Metabolism studies
  • Toxicity studies

…in a metabolically competent model of the human airway epithelium resembling an inflammatory state.

Endpoints/Outcome parameter

Calu-3 cells are cultured on permeable filter supports at the air liquid interface or under submersed conditions. LPS is added to the apical surface in order to stimulate a pro-inflammatory response, which is then quantified by measuring of cytokines (e.g. IL-6 and IL-8). Tight junction integrity is checked by measuring the transepithelial electrical resistance (TEER). The model is suitable as a screening tool to study the uptake and efficacy of different drug formulations by airway epithelial cells.

Readout parameter

  • Secretion of proinflammatory cytokines (e.g. IL-8, IL-4, IL-6, Il-1ß, TGF-alpha, RANTES, GM-CSF, MCP-1)
  • Transepithelial electrical resistance (TEER)
  • Mucus production
  • Cytotoxicity (e.g. WST-test, LDH release, ATP-content)
  • Gene expression (qRT-PCR, transcriptome analysis with Affymetrix GeneChip® )
  • Absorption and metabolism of the test compound (chemical analysis)

Quality management and validation

  • Internal quality management;
  • Controls
  • Blinded data collection and evaluation
  • Randomization
  • Others (e.g. historic controls)

References

  • Hansen, T. Knebel, J., Calu-3 cells as a model to estimate the pulmonary absorption of inhaled drugs for the treatment of COPD. In: The Toxicologist 51 (2012), 126/1, page 397, Abstract 1848