The role of epithelial cells and fibroblasts in the pathogenesis of chronic rhinosinusitis

PhD ThesisChronic rhinosinusitis without nasal polyps (CRSsNP) is a heterogeneous condition with common symptoms, clinical and radiological findings. CRSsNP is typified by inflammation of the sinonasal epithelium and development of fibrosis, yet its precise pathophysiology remains elusive. Recently stromal cells have been shown to act like immune effector cells in orchestrating chronic inflammation. Histological analysis of tissue biopsies from patients with CRSsNP demonstrates recruitment of circulating inflammatory cells, though the precise role of structural cells such as epithelial and fibroblast cells in CRSsNP remains to be discovered. Aims 1. (a) Recruit phenotyped cohorts of control & CRSsNP participants. (b) Characterise recruited CRSsNP participants’ tissue samples and isolated epithelial & fibroblast cells. 2. Assay the sinonasal environment to determine any association between, infection, inflammation and remodelling. 3. Identify clusters of genes differentially expressed in CRSsNP & control participants. Methods Cohorts of healthy control and CRSsNP participants were recruited. Matched tissue biopsy, epithelial and fibroblast cells were harvested together with clinical, radiological, microbiological and mucosal swab data. Tissue and cellular samples were characterised to confirm their identity and disease status. The sinonasal environment was characterised from mucosal swabs and analysed for a range of 40 human disease biomarkers. Transcriptome analysis was performed using microarrays and RNA sequencing with downstream bioinformatics investigation of the data. Results 47 age and sex matched CRSsNP and control participants were recruited, differing significantly in symptom and radiological scores. Histological analysis of tissue biopsy specimens was consistent with CRSsNP and control samples. Matched epithelial and fibroblast cells were generated. Assay of the sinonasal microenvironment identified 13 discriminant mediators separating CRSsNP samples from controls using a novel, non-invasive technique. Transcriptomics identified 239 differentially expressed genes in CRSsNP tissue biopsy samples. Cellular samples differed significantly from their matched tissue biopsies. Conclusions This thesis characterises a cohort of tightly defined CRSsNP patients and healthy controls to investigate the potential role of epithelial and fibroblast cells in CRSsNP. Transcriptomics has demonstrated clusters of genes upregulated in CRSsNP, however changes were not consistent in matched cellular samples questioning the validity of cellular models in CRSsNP. Additionally, a straightforward, non-invasive measure of the CRSsNP cytokine profile has been demonstrated. The mediators identified in these assays could potentially be developed as biomarkers of sinonasal inflammation as an adjunct in patient management.Wellcome Trus

Molecular mechanisms underlying the pathogenesis of nasal polyposis and its response to steroid treatment

Ph.DDOCTOR OF PHILOSOPH

An immunological perspective of the mucosal inflammation in chronic rhinosinusitis – lymphoid neo-organogenesis and humoral immunity

Chronic rhinosinusitis (CRS) is defined by the symptomatic inflammation of the nose and paranasal sinuses longer than 12 weeks. These symptoms include nasal discharge, nasal obstruction, facial pain and pressure, leading to a substantial impact on the quality of life of CRS patients. CRS can be phenotypically classified into either CRS without nasal polyps (CRSsNP) or CRS with nasal polyps (CRSwNP), based on the presence of endoscopically visualized nasal polyps in the middle meatus. Interestingly, ectopic accumulations of lymphoid cells are often observed within the nasal polyps of CRSwNP. This raises the question as to whether these aberrant lymphoid cell aggregates play a role in orchestrating the perpetual inflammation in CRS. Studies in the past have identified the increased amount of local class-switched antibodies in nasal polyps, but few studies have investigated the source of these immunoglobulins and utilized specific markers to study the presence of the organized lymphoid structures and their relation to disease severity in the context of CRS. This thesis investigates the significance of organized lymphoid neo-organogenesis in CRS pathogenesis and its effect on humoral immunity within both CRSsNP and CRSwNP patients.Thesis (Ph.D.) (Research by Publication) -- University of Adelaide, Adelaide Medical School, 201

Genetics and Epigenetics of Nasal Polyposis: A Systematic Review

[EN] Chronic rhinosinusitis (CRS) is an inflammatory disease of the nose and paranasal sinuses that is often associated with nasal polyposis (CRSwNP) in the most severe cases. As in other complex diseases, genetic factors are thought to play an important role in the risk and development of the disease. Environment may also modulate the epigenetic signature in affected patients. In the present systematic review, we aimed to compile all published data on genetic and epigenetic variations in CRSwNP since 2000. We found 104 articles, 24 of which were related to epigenetic studies. We identified more than 150 genetic variants in 99 genes involved in the pathogenesis of nasal polyposis. These were clustered into 8 main networks, linking genes involved in inflammation and immune response (eg, MHC), cytokine genes (eg, TNF), leukotriene metabolism, and the extracellular matrix. A total of 89 miRNAs were also identified; these are associated mainly with biological functions such as the cell cycle, inflammation, and the immune response. We propose a potential relationship between genes and the miRNAs identified that may open new lines of investigation. An in-depth knowledge of gene variants and epigenetic traits could help us to design more tailored treatment for patients with CRSwNP.Thematic Network of Cooperative Research in Health - RETICS (Red temática de investigación en salud Asma, Reacciones Adversas y Alérgicas, ARADYAL) of the Instituto de Salud Carlos II

Estudio de IL5RA como biomarcador genético en Poliposis Nasal

[ES] La IL-5 se caracteriza por su actividad en distintas células diana, entre las que se incluyen los linfocitos B, eosinófilos o basófilos. Es producida por linfocitos T, sobre todo Th2, ILC2 y mastocitos, y ejerce efectos en la proliferación y diferenciación, a través de su receptor (IL-5R), un heterodímero compuesto por una subunidad α específica de ligando (IL-5Rα) y otra subunidad β, común a otros receptores de citocinas. Debido a la relación de la poliposis nasal con la inflamación T2, la evolución de la poliposis nasal asociada al asma con eosinofilia se ha considerado un biomarcador para predecir la respuesta a los anticuerpos antiIL-5 en el asma. La terapia antagonista de IL-5 ha demostrado ser de utilidad en pacientes asmáticos con un fenotipo T2, siendo por lo tanto extensible el potencial de dicho tratamiento al de la inflamación en RSCcPN. La hipótesis principal de este estudio es que los pacientes con RSCcPN, asociados o no a otras patologías de vías respiratorias, atopia, hábitos tóxicos y a más variables clínicopatológicas, presentan una sobreexpresión del gen IL5RA respecto a los controles. La hipótesis secundaria se extrapola de la anterior, y es la posibilidad de considerar IL5RA como un biomarcador de poliposis nasal que pueda ser útil tanto en el diagnóstico, como en el seguimiento y el tratamiento, facilitando una terapia dirigida a determinados grupos de pacientes. En el estudio se plantearon los siguientes objetivos: 1. Determinar los niveles de expresión del gen IL5RA en sangre periférica en una población de pacientes con poliposis nasal frente a un grupo control. 2. Analizar las diferencias de expresión entre distintos fenotipos clínicos. 3. Determinar los niveles de expresión de IL5RA en tejido polipoideo y estudiar su relación con los niveles de expresión en sangre periférica. 4. Estudiar la posible correlación entre la expresión de IL5RA y otros genes que caracterizan grupos patológicos de rinosinusitis crónica, asma y otras enfermedades alérgicas

European position paper on rhinosinusitis and nasal polyps 2020

The European Position Paper on Rhinosinusitis and Nasal Polyps 2020 is the update of similar evidence based position papers published in 2005 and 2007 and 2012. The core objective of the EPOS2020 guideline is to provide revised, up-to-date and clear evidence-based recommendations and integrated care pathways in ARS and CRS. EPOS2020 provides an update on the literature published and studies undertaken in the eight years since the EPOS2012 position paper was published and addresses areas not extensively covered in EPOS2012 such as paediatric CRS and sinus surgery. EPOS2020 also involves new stakeholders, including pharmacists and patients, and addresses new target users who have become more involved in the management and treatment of rhinosinusitis since the publication of the last EPOS document, including pharmacists, nurses, specialised care givers and indeed patients themselves, who employ increasing self-management of their condition using over the counter treatments. The document provides suggestions for future research in this area and offers updated guidance for definitions and outcome measurements in research in different settings. EPOS2020 contains chapters on definitions and classification where we have defined a large number of terms and indicated preferred terms. A new classification of CRS into primary and secondary CRS and further division into localized and diffuse disease, based on anatomic distribution is proposed. There are extensive chapters on epidemiology and predisposing factors, inflammatory mechanisms, (differential) diagnosis of facial pain, allergic rhinitis, genetics, cystic fibrosis, aspirin exacerbated respiratory disease, immunodeficiencies, allergic fungal rhinosinusitis and the relationship between upper and lower airways. The chapters on paediatric acute and chronic rhinosinusitis are totally rewritten. All available evidence for the management of acute rhinosinusitis and chronic rhinosinusitis with or without nasal polyps in adults and children is systematically reviewed and integrated care pathways based on the evidence are proposed. Despite considerable increases in the amount of quality publications in recent years, a large number of practical clinical questions remain. It was agreed that the best way to address these was to conduct a Delphi exercise . The results have been integrated into the respective sections. Last but not least, advice for patients and pharmacists and a new list of research needs are included

European Position Paper on Rhinosinusitis and Nasal Polyps 2020

The European Position Paper on Rhinosinusitis and Nasal Polyps 2020 is the update of similar evidence based position papers published in 2005 and 2007 and 2012. The core objective of the EPOS2020 guideline is to provide revised, up-to-date and clear evidence-based recommendations and integrated care pathways in ARS and CRS. EPOS2020 provides an update on the literature published and studies undertaken in the eight years since the EPOS2012 position paper was published and addresses areas not extensively covered in EPOS2012 such as paediatric CRS and sinus surgery. EPOS2020 also involves new stakeholders, including pharmacists and patients, and addresses new target users who have become more involved in the management and treatment of rhinosinusitis since the publication of the last EPOS document, including pharmacists, nurses, specialised care givers and indeed patients themselves, who employ increasing self-management of their condition using over the counter treatments. The document provides suggestions for future research in this area and offers updated guidance for definitions and outcome measurements in research in different settings. EPOS2020 contains chapters on definitions and classification where we have defined a large number of terms and indicated preferred terms. A new classification of CRS into primary and secondary CRS and further division into localized and diffuse disease, based on anatomic distribution is proposed. There are extensive chapters on epidemiology and predisposing factors, inflammatory mechanisms, (differential) diagnosis of facial pain, allergic rhinitis, genetics, cystic fibrosis, aspirin exacerbated respiratory disease, immunodeficiencies, allergic fungal rhinosinusitis and the relationship between upper and lower airways. The chapters on paediatric acute and chronic rhinosinusitis are totally rewritten. All available evidence for the management of acute rhinosinusitis and chronic rhinosinusitis with or without nasal polyps in adults and children is systematically reviewed and integrated care pathways based on the evidence are proposed. Despite considerable increases in the amount of quality publications in recent years, a large number of practical clinical questions remain. It was agreed that the best way to address these was to conduct a Delphi exercise. The results have been integrated into the respective sections. Last but not least, advice for patients and pharmacists and a new list of research needs are included.Peer reviewe