Determining the effects of airborne particulates on immune and barrier epithelial cell function.

Abstract:
Approximately 24% of the particulate matter that humans breathe in each day consists of biological substances such as bacteria and fungi. These biological particulates make up the earth's microbiome. The microbiome plays important roles in human health and spread of disease as well as environmental processes, such as the water cycle. The microbiome is highly susceptive to the
environment and has been shown to be significantly different depending on the environment and bioaerosols sources present. Most research in to bioaerosols is on non-biological components, or of infectious agents in high risk environments. Whereas this study, will focus mainly on non-pathogenic bacteria and fungi. Although, the exact extent of how airborne biological pollutants impacts human health is unknown. Recent studies have shown that there are significant health risks associated with
particulate matter, as both biological and non-biological airborne particulates can trigger damage response pathways and lead to an immune response.
The immune response is comprised of extensive mechanisms that protect the host from infectious agents. Two major components of the innate immune response are epithelial cells that provide a physical barrier against microorganisms and macrophages, that aid in killing of pathogens, as well as the initiation of a greater more specific immune response. When these cells come into contact with pathogenic organisms, it triggers a damage response and leads to the activation of the inflammasome.
The inflammasome is a multimeric protein complex that is formed due to the presences of foreign or damage associated molecules. This leads to the activation of proteins within the macrophages and epithelial cells that combine in order to, secrete pro-inflammatory cytokines and cause pyroptosis, a form of inflammatory cell death, leading to the release of inflammatory mediators and the subsequent death of the invading microorganism. It has been shown in the previous studies that pathogenic bacteria, fungi and viruses can induce inflammasome activation. However, it is unknown whether non2 pathogenic bacteria and other airborne biological particulates can initiate the activation of the inflammasome and lead to an inflammatory state within the host. Therefore, determining what is present within the air with breathe in and how it affects the immune response is crucial and will be the aim of this project.

In order to examine the microbiome and its effect on immune cells and epithelial cells. Particulate matter samples will be recorded in different environments, during different seasons, where the biological component of the samples will be collected onto a filter. A novel technique will be developed in order to remove the biological contents from the filters, so identification of what is present can occur. With the extracted biological material examined using sequencing techniques such as 18s/16s and illumina. This will used to identify what is present within the atmosphere. Macrophages and epithelial cells will then be cultured with the biological components that have been collected. Expression of proteins associated with formation of the inflammasome will be examined by immunofluorescence and wester blotting, and expression of cell surface markers by flow cytometry.
Whilst expression of cytokines associated with production of the inflammasome and are known mediators of an inflammatory immune response will be examined by ELISA. These tests will allow for the analysis of the immune response in relation to the airborne particulates and will act as an indicator as to how the environment is affecting our health.

Aerosol science has a significant impact on a broad range of disciplines, extending from inhaled drug delivery, to combustion science and its health impacts, aerosol assisted routes to materials, climate change, and the delivery of agricultural and consumer products. Estimates of the global aerosol market size suggest it will reach $84 billion/year by 2024 with products in the personal care, household, automotive, food, paints and medical sectors. Air pollution leads to an estimated 30-40,000 premature deaths each year in the UK, and aerosols transmit human and animal infections. More than 12 million people in the UK live with lung disease such as asthma, and the NHS spends ~£5 billion/year on respiratory therapies. Many of the technological, societal and health challenges central to these areas rely on core skills and knowledge of aerosol science. Despite this, an Industrial Workshop and online survey (held in preparation for this bid) highlighted the current doctoral skills gap in aerosol science in the UK. Participating industries reported that only 15% of their employees working with aerosol science at doctoral-level having received any formal training. A CDT in aerosol science, CAS, will fill this skills gap, impacting on all areas of science where core training in aerosol science is crucial.

Impact on the UK aerosol community: Aerosol scientists work across governmental policy, industrial research and innovation, and in academia. Despite the considerable overlap in training needs for researchers working in these diverse sectors, current doctoral training in aerosol science is fragmentary and ad hoc (e.g. the annual Fundamentals of Aerosol Science course delivered by the Aerosol Society). In addition, training occurs within the context of individual disciplines, reinforcing artificial subject boundaries. CAS will bring coherence to training in the core physical and engineering science of aerosols, catalysing new synergies in research, and providing a focal point for training a multidisciplinary community of researchers. Working with the Aerosol Society, we will establish a legacy by providing training resources for future researchers through an online training portal.

Impact on industry and public-sector partners: 45 organisations have indicated they will act as CAS partners with interests in respiratory therapies, public health, materials manufacturing, consumer and agricultural products, instrumentation, emissions and environment. Establishing CAS will deliver researchers with the necessary skills to ensure the UK establishes and sustains a scientific and technical lead in their sectors. Further, it will provide an ideal mechanism for delivering Continuing Professional Development for the existing workforce practitioners. The activity of CAS is aligned to the Industrial Strategy Challenge Fund (e.g. through developing new healthcare technologies and new materials) and the EPSRC Prosperity Outcomes of a productive, healthy (e.g. novel treatments for respiratory disease) and resilient (e.g. adaptations to climate change, air quality) nation, with both the skilled researchers and their science naturally translating to long-lasting impact. Additionally, rigorous training in responsible innovation and ethical standards will lead to aerosol researchers able to contribute to developing: regulatory standards for medicines; policy on air quality and climate geoengineering; and regulations on manufactured nano-materials.

Public engagement: CAS will provide a focal point for engaging the public on topics in aerosol science that affect our daily lives (consumer products, materials) through to our health (inhalation therapeutics, disease transmission and impacts of pollution) and the future of our planet (geoengineering). Supported by a rigorous doctoral level training in aerosol science, this next generation of researchers will be ideally positioned to lead debates on all of these societal and technological challenges.