In Cystic Fibrosis (CF), lung disease arises due to multiple layers of dysfunction (infection, inflammation, and mucus dehydration). Impairment of bronchial epithelial cells, which play a major role in airway surface liquid modulation, and of phagocytes, including macrophages, which play a major role in the inflammatory response and bacterial clearance, both contribute to CF-related immune dysfunctions. Little is known about the mechanisms mediating the crosstalk between the cystic fibrosis transmembrane regulator (CFTR) protein, which is responsible for CF when mutated, and receptors involved in the immune regulation of the cells. We have identified a pathway that is disrupted in CF macrophages and interferes with the tightly regulated innate immune response to inflammatory triggers. We have discovered that CF macrophages fail to induce expression of the scaffold protein caveolin 1 (CAV1) in response to LPS and Pseudomonas aeruginosa, which, ultimately, leads to impaired negative regulation of Toll like receptor 4 (TLR4) signaling. More recently, we found that decreased CAV1 expression is due to high levels of microRNA-199a-5p, which persists in CF macrophages during TLR4 signaling, and targets CAV1. Furthermore, high levels of miR-199a-5p in CF M?s are due to blunted phosphatidylinositol 3-kinase (PI3K)/protein kinase B-1 (Akt1) signaling in response to inflammatory triggers. Now we have preliminary data showing that this pathway is also dysfunctional in CF epithelium. Thus, we have identified a novel dysregulated cellular pathway in CF that affects immune regulation. We hypothesize that CFTR mediates the assembly of a regulatory platform at the plasma membrane, which is necessary for the induction of the protective PI3K/Akt1/miR199-5p/CAV1 pathway in response to activation of TLRs. Moreover, inefficient downstream induction of this protective pathway in response to inflammation and infection reduces the ability of CF cells to respond to inflammatory stressors, leading to persistent hyper-inflammation, reduced anti-oxidative response, and, ultimately, the inability of the cells to re-establish cellular homeostasis and to survive. In this proposal we will: characterize the mechanism/s by which miR-199a-5p levels are dysregulated in CF cells (Aim 1); investigate the mechanism/s by which lack of functional CFTR alters Akt1/miR-199a-5p axis induction during TLR4/MyD88 signaling (Aim 2); test drugs that modulate the AKT/miR-199a/CAV1 pathway (Aim 3). Our findings have the potential to identify new molecular targets for therapeutic intervention in CF cells, which could halt the progression of chronic fatal airway hyper-inflammation and infection and delay lung deterioration in CF patients.

Public Health Relevance

Lung disease, characterized by bacterial infection and strong inflammation, is the major cause of death in patients with Cystic Fibrosis (CF). We have identified a novel cellular pathway that regulates the response to inflammatory stimuli, and is disrupted in cells of CF patients and in mouse models of the disease. Here, we will elucidate the mechanism that leads to dysfunction of this pathway in CF, which ultimately will determine specific molecular targets that can be used in novel therapeutic approaches to treat CF.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL123851-04
Application #
9747972
Study Section
Lung Cellular, Molecular, and Immunobiology Study Section (LCMI)
Program Officer
Lachowicz-Scroggins, Marrah Elizabeth
Project Start
2016-09-01
Project End
2020-06-30
Budget Start
2019-07-01
Budget End
2020-06-30
Support Year
4
Fiscal Year
2019
Total Cost
Indirect Cost
Name
Yale University
Department
Pediatrics
Type
Schools of Medicine
DUNS #
043207562
City
New Haven
State
CT
Country
United States
Zip Code
06520
Jin, Huiyan; Ciechanowicz, Andrzej K; Kaplan, Alanna R et al. (2018) Surfactant protein C dampens inflammation by decreasing JAK/STAT activation during lung repair. Am J Physiol Lung Cell Mol Physiol 314:L882-L892
Di Pietro, Caterina; Zhang, Ping-Xia; O'Rourke, Timothy K et al. (2017) Ezrin links CFTR to TLR4 signaling to orchestrate anti-bacterial immune response in macrophages. Sci Rep 7:10882