Posters

      Human In Vitro Models for Respiratory Toxicology: Evaluation of Goblet Cell Hyperplasia, Mucus Production, and Ciliary Beating Assays

      Frentzel, S.; Aragon, M.; Hoeng, J.; Ito, S.; Ishikawa, S.; Budde, J.; Maione, A.; Hayden, P.; Fields, W.; Keyser, B.; Haswell, L.; Azzopardi, D.; Behrsing , H.

      Conference date
      Mar 15, 2018
      Conference name
      Society of Toxicology (SOT) 2018
      Link to more information
      Link to more information
      Topic
      Summary

      Robust non-animal models and assays for pulmonary toxicology are required to make competent product development and risk assessments for new materials requiring toxicity testing. Three in vitro assays (goblet cell hyperplasia [GCH], ciliary beat frequency [CBF], and MUC5AC quantitation) were evaluated for performance and reproducibility. To assess these assays, 6 laboratories contributed data using a common protocol utilizing IL-13 as an inducer of adverse mucociliary-relevant tissue changes. MatTek EpiAirway™ and Epithelix MucilAir™ 3D tissue models were used to evaluate endpoints using histology for GCH, software-based applications, Cilia FA and SAVA, for CBF, and ELISA assay for MUC5AC. Continuous 10 ng/mL IL-13 (GCH, MUC5AC) exposures or one hour 10 μM procaterol (CBF) exposures prior to day 7 and 14 time-points were included as positive controls. Quality control endpoints (e.g. adenylate kinase tissue content and trans-epithelial electrical resistance) were also evaluated. Multi-fold increases (ranging from 2.6 to 33-fold, and 1.5 to 238-fold) in MUC5AC-stained goblet cells were measured in both tissue models after exposure with IL-13 after 7 and 14 days induction, respectively. For CBF, procaterol caused a significant increase, and IL-13 elicited a significant decrease as expected. However, the MUC5AC ELISA did not yield consistent results when frozen apical rinse samples were thawed and assayed. These results suggest these non-animal test systems may provide consistent, human-relevant data corresponding to key events involved in respiratory disease. A streamlined protocol using these controls will be applied toward additional testing. These assays, utilized in a pragmatic manner with other in vitro assays have the potential to be included in a Reduced Risk Product assessment framework.

      PMIScience.com is operated by Philip Morris International for the purpose of publishing and disseminating scientific information about Philip Morris International’s efforts in support of its smoke-free product portfolio. This site is a global site for use by scientists, the public health and regulatory communities, and other stakeholders with an interest in tobacco policy. The purpose of this site is not advertising or marketing, nor is it directed at any specific market. It is not intended for use by consumers. New tobacco products sold in the United States are subject to FDA regulation; therefore the content of this site is not intended to make, and nor should it be construed as making, any product related claims in the United States without proper FDA authorization. ​
      ​
      Reduced Risk Products ("RRPs”) is the term we use to refer to products that present, are likely to present, or have the potential to present less risk of harm to smokers who switch to these products versus continuing smoking. PMI has a range of RRPs in various stages of development, scientific assessment and commercialization. All of our RRPs are smoke-free products that deliver nicotine with far lower quantities of harmful and potentially harmful constituents than found in cigarette smoke.