In this renewal application, we describe a new pathway signaling neutrophil (PMN) influx and damage to the airways that was delineated in the parent grant and that may play a role as an initiator or cofactor for chronic lung diseases such as obstructive pulmonary disease (COPD), cystic fibrosis (CF), and chronic lung transplant rejection. Specifically, chemical or enzymatic breakdown of collagen releases a tripeptide, PGP, and/or a related PGP-containing sequence that is chemotactic for PMN in vitro. We demonstrate that introduction of PGP into the airways causes a robust influx of PMN, but not monocytes. Remarkably, the PMN chemotactic activity of PGP may be due to a marked structural relatedness to a receptor binding domain of ELR+ CXC chemokines such as IL-8 which contain this collagen sequence or a close analog. Prolonged airway exposure to this peptide causes alveolar enlargement and right ventricular hypertrophy and thus recapitulates aspects of COPD. We have developed a potent antagonist, Arg-Thr-Arg, termed RTR, which binds PGP in solution and blocks its in vitro chemotactic activity and in vivo pathophysiologic effects. Likely as a result of shared structure with PGP, RTR also blocks IL-8 activity. In a mouse model of LPS-induced COPD, RTR completely inhibited alveolar enlargement and right ventricular hypertrophy. The degradation of collagen and release of PGP is shown to be a stepwise process initially involving matrix metalloproteases (MMP)-8 and/or 9 with prolyl endopeptidase (PE) catalyzing the final reaction. We have found that a highly specific PE inhibitor, N-benzyloxycarbonyl (Z) prolyl prolinal (ZPP), can block airway PGP production as well as PMN influx in response to LPS. Surprisingly, the very potent blunting of PMN influx by ZPP appears to be due not only to PE inhibition but also as a result of ZPP antagonism of CXCR 1/2 likely as a result of structural similarity between ZPP and PGP. This suggests the intriguing possibility, to be explored in this proposal, that a single compound can be developed that is both a protease inhibitor and receptor antagonist. If successful, this would represent an entirely new chimeric type of lead compound for the treatment of neutrophilic inflammation, in general, and lung diseases in particular. We have found that PGP is present in bronchoalveolar lavage fluids (BALF) and/or sputum from virtually all CF, COPD or chronic lung transplant rejection patients but not controls or asthmatics. Furthermore, sputum from COPD and CF patients but not control individuals contains all the enzymatic machinery necessary for the ex vivo generation of PGP from purified collagen and such PGP production can be blocked by the PE inhibitor, ZPP. Collectively, these findings lead us to hypothesize that PGP and PE represent novel biomarkers for COPD that may contribute to disease and that PGP and PE represent attractive therapeutic targets at the level of the ligand, receptor, and generating enzyme.

Public Health Relevance

COPD is now the fourth leading cause of death in the U.S. and is expected to be the third by 2020. This disease causes much human suffering as well as a large monetary burden. An understanding of COPD that leads to successful therapeutics has been hampered by a lack of biomarkers to diagnose and use as endpoints in clinical trials as well as a lack of a fundamental understanding of the disease process itself. If successful, the current proposal will yield fundamental insights into the disease process and a new and specific biomarker for COPD as well as suggesting new therapeutic targets.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL077783-08
Application #
8451420
Study Section
Lung Cellular, Molecular, and Immunobiology Study Section (LCMI)
Program Officer
Punturieri, Antonello
Project Start
2006-04-01
Project End
2014-03-31
Budget Start
2013-04-01
Budget End
2014-03-31
Support Year
8
Fiscal Year
2013
Total Cost
$345,227
Indirect Cost
$109,577
Name
University of Alabama Birmingham
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
063690705
City
Birmingham
State
AL
Country
United States
Zip Code
35294
Hahn, Cornelia S; Scott, David W; Xu, Xin et al. (2015) The matrikine N-α-PGP couples extracellular matrix fragmentation to endothelial permeability. Sci Adv 1:
Wells, J Michael; Gaggar, Amit; Blalock, J Edwin (2015) MMP generated matrikines. Matrix Biol 44-46:122-9
Abdul Roda, Mojtaba; Sadik, Mariam; Gaggar, Amit et al. (2014) Targeting prolyl endopeptidase with valproic acid as a potential modulator of neutrophilic inflammation. PLoS One 9:e97594
Koelink, Pim J; Overbeek, Saskia A; Braber, Saskia et al. (2014) Collagen degradation and neutrophilic infiltration: a vicious circle in inflammatory bowel disease. Gut 63:578-87
Kong, Michele Y F; Clancy, John P; Peng, Ning et al. (2014) Pulmonary matrix metalloproteinase-9 activity in mechanically ventilated children with respiratory syncytial virus. Eur Respir J 43:1086-96
Le Jan, Sébastien; Plée, Julie; Vallerand, David et al. (2014) Innate immune cell-produced IL-17 sustains inflammation in bullous pemphigoid. J Invest Dermatol 134:2908-17
Wells, J Michael; O'Reilly, Philip J; Szul, Tomasz et al. (2014) An aberrant leukotriene A4 hydrolase-proline-glycine-proline pathway in the pathogenesis of chronic obstructive pulmonary disease. Am J Respir Crit Care Med 190:51-61
O'Reilly, Philip J; Jackson, Patricia L; Wells, J Michael et al. (2013) Sputum PGP is reduced by azithromycin treatment in patients with COPD and correlates with exacerbations. BMJ Open 3:e004140
Raju, S Vamsee; Jackson, Patricia L; Courville, Clifford A et al. (2013) Cigarette smoke induces systemic defects in cystic fibrosis transmembrane conductance regulator function. Am J Respir Crit Care Med 188:1321-30
Hardison, Matthew Thomas; Blalock, James Edwin (2012) Molecular recognition theory and sense-antisense interaction: therapeutic applications in autoimmunity. Front Biosci (Elite Ed) 4:1864-70

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