Pseudomonas aeruginosa is the major respiratory pathogen in the pathogenesis of Cystic Fibrosis (CF) and the ineffective immune response to this pathogen is thought to cause the majority of the lung damage characteristic of this disease. In the later stages of CF, P. aeruginosa reside in biofilm communities in the lung, accounting for their resistance to antibiotic therapies. To date, little is known about host factors that promote the transition of P. aeruginosa from acute to chronic infection in CF. It has been reported that CF patients show a reduced ability to clear P. aeruginosa acquired during respiratory viral infections and 85% of new pseudomonal colonization in CF patients followed a respiratory viral infection within 3 weeks. We have demonstrated that virus co-infection, and the subsequent antiviral type III interferon response, promote biofilm conversion by P. aeruginosa. Type III interferon has potent antiviral activity, but in addition, interferon stimulated gene (ISG) effecto functions have been reported to promote pathogen replication, suggesting that pathogens have evolved to subvert and even benefit from the interferon response. A fundamental aspect in microbial interactions is the relentless battle for nutrients, including iron, where the host makes every effort to restrict access to pathogens and the pathogens have developed sophisticated strategies to acquire the nutrients they require for growth from the host. Using a unique model to culture P. aeruginosa biofilms in association with human CF airway epithelial cells, we will examine whether the host response to viral infection, namely type III interferon signaling, promotes biofilm conversion by P. aeruginosa through a mechanism of inappropriate iron secretion. To this end, we will (1) define the iron regulatory pathways that are altered by virus infection and type III interferon signaling in the airway epithelium, (2) define the mechanism by which iron is mobilized into the airway surface liquid and how P. aeruginosa acquires host iron and (3) define a role for the ISG, viperin/RSAD2, in promoting iron secretion from airway epithelial cells to enhance bacterial biofilm growth. The proposed experiments would provide a novel link between viral co-infection and the establishment of chronic bacterial colonization, with important implications in the progression of CF lung disease. Our goal is to elucidate the molecular mechanism for virus-stimulated bacterial biofilms and thus, identify new targets that could delay acquisition and chronic bacterial colonization, or work in conjunction with existing therapies, to eradicate P. aeruginosa in CF patients.

Public Health Relevance

Pseudomonas aeruginosa is the major cause of morbidity and mortality for patients with Cystic Fibrosis (CF). Using a unique model system to culture P. aeruginosa biofilms in association with human CF airway epithelial cells, we have made the novel observation that virus co-infections promote chronic infection by P. aeruginosa. The studies proposed in this application will elucidate the molecular mechanism for viral-bacterial co-infections in the CF airway and likely identify new therapeutic targets to delay acquisition and chronic bacterial colonization, or work in conjunction with existing therapies, to eradicate P. aeruginosa in CF patients.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL123771-02
Application #
9041677
Study Section
Lung Cellular, Molecular, and Immunobiology Study Section (LCMI)
Program Officer
Caler, Elisabet V
Project Start
2015-09-01
Project End
2019-07-31
Budget Start
2016-08-01
Budget End
2017-07-31
Support Year
2
Fiscal Year
2016
Total Cost
Indirect Cost
Name
University of Pittsburgh
Department
Genetics
Type
Schools of Medicine
DUNS #
004514360
City
Pittsburgh
State
PA
Country
United States
Zip Code
15213
Hvorecny, Kelli L; Dolben, Emily; Moreau-Marquis, Sophie et al. (2018) An epoxide hydrolase secreted by Pseudomonas aeruginosa decreases mucociliary transport and hinders bacterial clearance from the lung. Am J Physiol Lung Cell Mol Physiol 314:L150-L156
Kiedrowski, Megan R; Gaston, Jordan R; Kocak, Brian R et al. (2018) Staphylococcus aureus Biofilm Growth on Cystic Fibrosis Airway Epithelial Cells Is Enhanced during Respiratory Syncytial Virus Coinfection. mSphere 3:
Purcaro, Giorgia; Rees, Christiaan A; Melvin, Jeffrey A et al. (2018) Volatile fingerprinting of Pseudomonas aeruginosa and respiratory syncytial virus infection in an in vitro cystic fibrosis co-infection model. J Breath Res 12:046001
Melvin, Jeffrey A; Gaston, Jordan R; Phillips, Shawn N et al. (2017) Pseudomonas aeruginosa Contact-Dependent Growth Inhibition Plays Dual Role in Host-Pathogen Interactions. mSphere 2:
Flitter, Becca A; Hvorecny, Kelli L; Ono, Emiko et al. (2017) Pseudomonas aeruginosa sabotages the generation of host proresolving lipid mediators. Proc Natl Acad Sci U S A 114:136-141
Lashua, Lauren P; Melvin, Jeffrey A; Deslouches, Berthony et al. (2016) Engineered cationic antimicrobial peptide (eCAP) prevents Pseudomonas aeruginosa biofilm growth on airway epithelial cells. J Antimicrob Chemother 71:2200-7
Melvin, Jeffrey A; Lashua, Lauren P; Kiedrowski, Megan R et al. (2016) Simultaneous Antibiofilm and Antiviral Activities of an Engineered Antimicrobial Peptide during Virus-Bacterium Coinfection. mSphere 1:
Hendricks, Matthew R; Bomberger, Jennifer M (2016) Digging through the Obstruction: Insight into the Epithelial Cell Response to Respiratory Virus Infection in Patients with Cystic Fibrosis. J Virol 90:4258-4261
Hendricks, Matthew R; Lashua, Lauren P; Fischer, Douglas K et al. (2016) Respiratory syncytial virus infection enhances Pseudomonas aeruginosa biofilm growth through dysregulation of nutritional immunity. Proc Natl Acad Sci U S A 113:1642-7
Melvin, Jeffrey A; Bomberger, Jennifer M (2016) Compromised Defenses: Exploitation of Epithelial Responses During Viral-Bacterial Co-Infection of the Respiratory Tract. PLoS Pathog 12:e1005797

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