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Regulation of Type-2 Airway Inflammation in Cystic Fibrosis and Asthma

Donnell, Mason (2020) Regulation of Type-2 Airway Inflammation in Cystic Fibrosis and Asthma. Doctoral Dissertation, University of Pittsburgh. (Unpublished)

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Asthma, chronic obstructive pulmonary disease (COPD) and cystic fibrosis (CF) patients can all experience type-2 airway inflammation. The aim of this research was to understand how type-2 airway inflammation is regulated in models of CF and asthma by the receptor for advanced glycation endproducts (RAGE). Previous studies indicate RAGE is crucial for induction of type-2 airway inflammation. Intriguingly, RAGE and type-2 proinflammatory cytokines are elevated in CF patient sputum and during infection with Pseudomonas aeruginosa. This led to the hypothesis that RAGE regulates IL-13 induced mucus production seen in CF patients.
Using a RAGE antagonist, this study shows RAGE regulates mucin gene MUC5AC expression, the expression of transcription factors regulating mucous metaplasia (CLCA1, SPDEF and FOXA3) and exotoxin-2 production in Non-CF and CF human bronchial epithelial cells (HBECs). By transfecting Non-CF and CF HBECs with dicer substrate siRNA (dsiRNA) to deplete RAGE expression, this study shows RAGE regulates these phenomena through activation of STAT6 upstream of mucous gene expression. CF HBECs were hyperresponsive to IL-13 treatment with elevated MUC5AC, CLCA1 and FOXA3 expression, which may be driven by sustained activation of STAT6. To test the role of RAGE in vivo, exotoxin derived from P. aeruginosa was used to model type-2 airway inflammation in WT and RAGE knock-out mice (RAGE-/-). WT animals treated with exotoxin were found to develop mucus production, though this was absent in RAGE-/- animals.
In a second study, RAGE regulation of IL-33 initiation of type-2 allergic airway inflammation (AAI) was investigated. Prior work shows WT mice exposed to allergens upregulate IL-33 in the lung, while RAGE-/- mice do not significantly upregulate the cytokine alarmin. To further understand how RAGE regulates IL-33 production, this study hypothesized RAGE expressing type-1 alveolar epithelial cells (ATIs) release IL-33 in response to allergens. After stimulation with Alternaria alternata (AA), RAGE expressing murine ATIs did not release IL-33. Human alveolar organoids were also tested but were unable to release IL-33 after stimulation with AA. Collectively, these studies show RAGE plays a role in IL-13 induced mucus production in CF HBECs and that ATIs are not solely responsible for IL-33 release.


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Item Type: University of Pittsburgh ETD
Status: Unpublished
CreatorsEmailPitt UsernameORCID
Donnell, Masonmld140@pitt.edumld140
ETD Committee:
TitleMemberEmail AddressPitt UsernameORCID
Committee ChairChu, Charleenctc4@pitt.eductc4
Thesis AdvisorOury, Timtdoury@pitt.edutdoury
Committee MemberAlder, Jonathanjalder@pitt.edujalder
Committee MemberKliment, Corrineckliment@pitt.educkliment
Committee MemberOrtiz, Luislao1@pitt.edulao1
Date: 2 September 2020
Date Type: Publication
Defense Date: 27 July 2020
Approval Date: 2 September 2020
Submission Date: 17 August 2020
Access Restriction: No restriction; Release the ETD for access worldwide immediately.
Number of Pages: 198
Institution: University of Pittsburgh
Schools and Programs: School of Medicine > Pathology
Degree: PhD - Doctor of Philosophy
Thesis Type: Doctoral Dissertation
Refereed: Yes
Uncontrolled Keywords: Asthma, Cystic Fibrosis, RAGE, Lung Inflammation, Type-2 Immunity, Th2 Inflammation
Date Deposited: 02 Sep 2020 20:08
Last Modified: 02 Sep 2020 20:08

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