Development and Testing of Pharmacological Tools to Evaluate the Role of Protease-Activated Receptor-2 in Allergic Asthma

Despite the variety of asthma pharmacotherapy options that exist, a large subset of asthma patients present with poorly controlled disease. Extensive studies supporting a central role for Protease-Activated Receptor-2 (PAR2) in allergic asthma pathophysiology have promoted drug development efforts towards this molecular target. I report here that PAR2 antagonist C391 blocks signaling and symptoms caused by the asthma causing allergen Alternaria alternata. I show C391 effectively inhibits A. alternata-induced PAR2-MAPK and -Ca2+ signaling in vitro and attenuates asthma symptom development (e.g. airway hyperresponsiveness, excessive mucus production, and inflammation) in a murine model of asthma. Use of in vitro studies enable the cell-specific examination of PAR2 contribution to A. alternata-induced airway inflammation. I show PAR2 signaling plays an important role in the initiation of asthma through the finding that PAR2 signaling is required for full cytokine and chemokine release from A. alternata-exposed epithelial cells when I compare release from parental cells and CRISPR-produced PAR2 knockout cells. C391 inhibition of PAR2 signaling in A. alternata-induced lymphocyte migration reveal PAR2 signaling is required for effective chemotaxis. Finally, I show development and screening of three novel PAR2 ligands, C732, C781 and C782. Cell impedance screening and PAR2 binding assessment highlight C781 as a lead compound for asthma therapy. Further PAR2 signaling characterization reveals C781 to be a PAR2-specific, biased MAPK antagonist. My studies demonstrate for the first time that a PAR2-specific antagonist provides pharmacological control of asthma in a pre-clinical model and presents novel lead compounds for improved drug development.

Name thesis:degree_name

Ph.D.

Level thesis:degree_level

doctoral

Discipline thesis:degree_discipline

Graduate College

Grantor dc:publisher

The University of Arizona.

Year dc:date.issued

2019

Author dc:creator

• Rivas, Candy Mavis

Advisor dc:contributor.advisor

• Boitano, Scott A.

Committee members dc:contributor.committeemember

• Burt, Janis M.
• Fregosi, Ralph F.
• Vanderah, Todd W.
• Ledford, Julie G.

• Airway
• Allergens
• Asthma
• Inflammation
• PAR2

Statement dc:rights

• Copyright © is held by the author. Digital access to this material is made possible by the University Libraries, University of Arizona. Further transmission, reproduction, presentation (such as public display or performance) of protected items is prohibited except with permission of the author.

Language dc:language.iso

en

Handle dc:identifier.uri

http://hdl.handle.net/10150/632588

OAI identifier oai:identifier

oai:repository.arizona.edu:10150/632588