Understanding Mechanisms Driving Lung Disease Caused by Environmental Particulate Matter

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Collectively, respiratory diseases are the leading cause of death worldwide. As a consequence respiratory research has increased by 500% and animal models have supported approximately 20% of this research. More than 900,000 mice have been used in the last decade alone to help understand human respiratory health, however, the physiological and anatomic differences between the human and mouse lungs have made it difficult to translate research findings into human therapies. Inhalation research focuses on the effect of inhaled chemicals, particles, bacteria, viruses, drug delivery systems and pollution in both healthy lung and diseased lungs. The most widely used human specific models of the human lung are cell-based models that focus on lung epithelial cells, the cells that make up the surface of the lungs. While these models are useful to model the response of epithelium to inhaled substances, the whole lung response is extremely important and encompasses complex cell-to-cell interactions and signalling, immune cells and extracellular matrix modifications that currently only mouse models can fully replicate, hence their over-use.We will address this gap in model platforms by creating a multi-cell type, immune competent model of human lung alveoli that is animal-free, capable of stretching and applicable to many areas of respiratory research. To achieve this we will use stem cell derived type II alveoli epithelial cells with macrophages, fibroblasts and endothelial cells in a synthetic hydrogel developed by our industrial partner Manchester BIOGEL. We will demonstrate application of the model to understand how atmospheric particulate matter, the most significant environmental pollutant in the world, affects lung health. The objectives below will provide an opportunity for a "next-generation" model of the human lung alveoli, capable of modelling epithelial response, fibrosis as well as that of the immune system and could provide important mechanistic, diagnostic and therapeutic information across many areas of respiratory research. 1): Combine hIPSC derived lung cell types into synthetic animal-free hydrogels.We will bring together our stem cell derived type II alveoli epithelial cells, with macrophages, fibroblasts and endothelial cells in a synthetic hydrogel to generate a 3D multi-cell-type, immune competent organoid model of the human lung alveoli. We will characterise structure of the organoid in different hydrogel formulations that mimic fibrosis both with and without stretch forces. We will then compare and validate individual cell behaviour to human precision cut lung slices cultured under equivalent conditions. 2) Understand the impact of particulate matter on lung alveoli tissueTo further demonstrate application of the model we will expose the alveoli organoids to environmental particulate matter in different hydrogel and stretch characteristics identified in Obj 1. We will then characterise the response of each cell type and compare and validate the response using human precision cut lung slices as in Obj 1. 3) Educate and train new users in development and manipulation of hIPSC derived alveoli organoids.We will host a conference focussed on current animal free models of human health and disease to educate current and future research leaders in use and development of animal-free models of human health. We will also host a 1-week training course for researchers that would like to adopt and use our model for their own research purposes.Together these objective will create a "next generation" model of human alveoli tissue applicable that could reduce and replace animal models across many different respiratory diseases. Importantly, there will be training opportunities provided to other researchers that will allow them to adopt and modify the model to their own experimental needs, further enhancing the possibility that the platform will have a positive impact in reducing animal use in research.