Neutrophil-targeted therapy for severe asthma - PROJECT SUMMARY/ABSTRACT This Mentored Research Scientist Development Award (K01) will support Dr. Rosemary Bayless, DVM, MS, PhD, DACVIM (LAIM), as she establishes her independent research career studying novel therapies for neutrophilic asthma and other neutrophil-mediated diseases in human patients. Dr. Bayless’ specialized training in veterinary comparative medicine, equine internal medicine and inflammatory pathophysiology make her uniquely qualified to advance translational research by leveraging both experimental and spontaneous models of human inflammatory diseases. The proposed aims and training plan, focusing on the mechanisms of a novel therapeutic for neutrophilic asthma, were carefully designed to provide Dr. Bayless with advanced training in 3D in vitro models, co-culture systems, and preclinical murine models, as well as study design and grant writing. Through mentorship from basic and translational research experts, established physician- scientists, and an experienced biostatistician, Dr. Bayless will develop key skills that will help launch a successful research-intensive career as an independent R01-funded translational clinician-scientist. The proposed training project addresses asthma, a chronic, progressive, debilitating respiratory disease. Asthmatic patients with neutrophilic airway inflammation tend to have more severe and treatment-resistant clinical signs compared to those with eosinophilic asthma. Novel asthma therapies that target neutrophilic inflammation would benefit millions of patients around the world. Withaferin A (WFA), derived from Withania somnifera, is a promising drug candidate for neutrophilic asthma. WFA has beneficial effects in models of other respiratory diseases, and we have shown that WFA directly inhibits neutrophilic inflammation in vitro and mitigates airway neutrophilia in a neutrophilic asthma mouse model. However, therapeutic mechanism(s) of WFA are not well understood, and continued research is needed to elucidate cellular processes impacted by WFA. The research objective of this K01 is to investigate the mechanisms that underlie therapeutic effects of WFA for neutrophilic asthma, utilizing advanced 3D in vitro systems to model key steps in neutrophil recruitment, co- culture to model processes that are central to inflammation resolution, and a published mouse model of neutrophilic asthma to evaluate these processes in vivo. In Aim 1, we will establish the effect of WFA on neutrophil migration across lung endothelium and airway epithelium using microfluidic devices and air-liquid interface culture, respectively. In Aim 2, we will determine the effect of WFA on neutrophil efferocytosis by alveolar macrophages and on alveolar macrophage phenotype. In Aim 3, we will investigate whether WFA mitigation of airway inflammation in a mouse model of neutrophilic asthma is mediated by decreased neutrophil recruitment to the airway and/or increased neutrophil clearance from the airway. Successful completion of these aims will provide valuable research training for Dr. Bayless and will inform future therapeutic studies for neutrophilic asthma, both in a naturally occurring equine model of neutrophilic asthma and in human patients.
