Coagulation factor Xa appears involved in the pathogenesis of pulmonary fibrosis. Through its interaction with protease activated receptor-1, this protease signals myofibroblast differentiation in lung fibroblasts. Although fibrogenic stimuli induce factor X synthesis by alveolar cells, the mechanisms of local posttranslational factor X activation are not fully understood. Cell-derived microparticles are submicron vesicles involved in different physiological processes, including blood coagulation; they potentially activate factor X due to the exposure on their outer membrane of both phosphatidylserine and tissue factor. We postulated a role for procoagulant microparticles in the pathogenesis of interstitial lung diseases. Nineteen patients with interstitial lung diseases and 11 controls were studied. All subjects underwent bronchoalveolar lavage; interstitial lung disease patients also underwent pulmonary function tests and high resolution CT scan. Microparticles were enumerated in the bronchoalveolar lavage fluid with a solid-phase assay based on thrombin generation. Microparticles were also tested for tissue factor activity. In vitro shedding of microparticles upon incubation with H2O2 was assessed in the human alveolar cell line, A549 and in normal bronchial epithelial cells. Tissue factor synthesis was quantitated by real-time PCR. Total microparticle number and microparticle-associated tissue factor activity were increased in interstitial lung disease patients compared to controls (84±8 vs. 39±3 nM phosphatidylserine; 293±37 vs. 105±21 arbitrary units of tissue factor activity; mean±SEM; p<.05 for both comparisons). Microparticle-bound tissue factor activity was inversely correlated with lung function as assessed by both diffusion capacity and forced vital capacity (r2 = .27 and .31, respectively; p<.05 for both correlations). Exposure of lung epithelial cells to H2O2 caused an increase in microparticle-bound tissue factor without affecting tissue factor mRNA. Procoagulant microparticles are increased in interstitial lung diseases and correlate with functional impairment. These structures might contribute to the activation of factor X and to the factor Xa-mediated fibrotic response in lung injury.
References
[1]
American Thoracic Society/European Respiratory Society (2002) American Thoracic Society/European Respiratory Society International Multidisciplinary Consensus Classification of the Idiopathic Interstitial Pneumonias. Am J Respir Crit Care Med 165: 277–304.
[2]
Raghu G, Collard HR, Egan JJ, Martinez FJ, Behr J, et al. (2011) An official ATS/ERS/JRS/ALAT statement: idiopathic pulmonary fibrosis: evidence-based guidelines for diagnosis and management. Am J Respir Crit Care Med 183: 788–824.
[3]
Furie B, Furie BC (1992) Molecular and cellular biology of blood coagulation. N Engl J Med 326: 800–806.
[4]
Giesen PLA, Rauch U, Bohrmann B, Kling D, Roque M, et al. (1999) Blood-borne tissue factor: Another view of thrombosis. Proc Natl Acad Sci 96: 2311–2315.
[5]
Falati S, Liu Q, Gross P, Merrill-Skoloff G, Chou J, et al. (2003) Accumulation of Tissue Factor into Developing Thrombi In Vivo Is Dependent upon Microparticle P-Selectin Glycoprotein Ligand 1 and Platelet P-Selectin. J Exp Med 197: 1585–1598.
[6]
Celi A, Lorenzet R, Furie BC, Furie B (2004) Microparticles and a P-selectin-mediated pathway of blood coagulation. Dis Markers 20: 347–352.
[7]
Distler JH, Huber LC, Gay S, Distler O, Pisetsky DS (2006) Microparticles as mediators of cellular cross-talk in inflammatory disease. Autoimmunity 39: 683–690.
[8]
Freyssinet JM (2003) Cellular microparticles: what are they bad or good for? J Thromb Haemost 1: 1655–1662.
[9]
Hugel B, Martinez MC, Kunzelmann C, Freyssinet JM (2005) Membrane microparticles: two sides of the coin. Physiology (Bethesda) 20: 22–27.
[10]
Distler JHW, Distler O (2010) Inflammation: Microparticles and their roles in inflammatory arthritides. Nature Reviews Rheumatology 6: 385–386.
[11]
Neri T, Armani C, Pegoli A, Cordazzo C, Carmazzi Y, et al. (2011) Role of NF-kappaB and PPAR-gamma in lung inflammation induced by monocyte-derived microparticles. Eur Respir J 37: 1494–1502.
[12]
Bastarache JA, Fremont RD, Kropski JA, Bossert FR, Ware LB (2009) Procoagulant alveolar microparticles in the lungs of patients with acute respiratory distress syndrome. Am J Physiol Lung Cell Mol Physiol 297: L1035–41.
[13]
Imokawa S, Sato A, Hayakawa H, Kotani M, Urano T, et al. (1997) Tissue factor expression and fibrin deposition in the lungs of patients with idiopathic pulmonary fibrosis and systemic sclerosis. Am J Respir Crit Care Med 156: 631–636.
[14]
Hernandez-Rodriguez NA, Cambrey AD, Harrison NK, Chambers RC, Gray AJ, et al. (1995) Role of thrombin in pulmonary fibrosis. Lancet 346: 1071–1073.
[15]
Howell DC, Goldsack NR, Marshall RP, McAnulty RJ, Starke R, et al. (2001) Direct thrombin inhibition reduces lung collagen, accumulation, and connective tissue growth factor mRNA levels in bleomycin-induced pulmonary fibrosis. Am J Pathol 159: 1383–1395.
[16]
Sprunger DB, Olson AL, Huie TJ, Fernandez-Perez ER, Fischer A, et al. (2012) Pulmonary fibrosis is associated with an elevated risk of thromboembolic disease. Eur Respir J 39: 125–132.
[17]
Kubo H, Nakayama K, Yanai M, Suzuki T, Yamaya M, et al. (2005) Anticoagulant therapy for idiopathic pulmonary fibrosis. Chest 128: 1475–1482.
[18]
Noth I, Anstrom KJ, Calvert SB, de Andrade J, Flaherty KR, et al. (2012) A placebo-controlled randomized trial of warfarin in idiopathic pulmonary fibrosis. Am J Respir Crit Care Med 186: 88–95.
[19]
Chambers RC (2008) Procoagulant signalling mechanisms in lung inflammation and fibrosis: novel opportunities for pharmacological intervention? Br J Pharmacol 153 Suppl 1S367–78.
[20]
Scotton CJ, Krupiczojc MA, Konigshoff M, Mercer PF, Lee YC, et al. (2009) Increased local expression of coagulation factor X contributes to the fibrotic response in human and murine lung injury. J Clin Invest 119: 2550–2563.
[21]
Miller MR, Crapo R, Hankinson J, Brusasco V, Burgos F, et al. (2005) General considerations for lung function testing. Eur Respir J 26: 153–161.
[22]
Neri T, Cordazzo C, Carmazzi Y, Petrini S, Balia C, et al. (2012) Effects of peroxisome proliferator activated receptors-gamma agonists on the generation of microparticles by monocytes/macrophages. Cardiovasc Res 11: 243–247.
[23]
Di Stefano R, Barsotti MC, Armani C, Santoni T, Lorenzet R, et al. (2009) Human peripheral blood endothelial progenitor cells synthesize and express functionally active tissue factor. Thromb Res 123: 925–930.
[24]
Celi A, Cianchetti S, Petruzzelli S, Carnevali S, Baliva F, et al. (1999) ICAM-1-independent adhesion of neutrophils to phorbol ester-stimulated human airway epithelial cells. Am J Physiol 277: L465–71.
[25]
Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods 25: 402–408.
[26]
Reid VL, Webster NR (2012) Role of microparticles in sepsis. Br J Anaesth 109: 503–513.
[27]
Cerri C, Chimenti D, Conti I, Neri T, Paggiaro P, et al. (2006) Monocyte/macrophage-derived microparticles up-regulate inflammatory mediator synthesis by human airway epithelial cells. J Immunol 177: 1975–1980.
[28]
Porro C, Lepore S, Trotta T, Castellani S, Ratclif L, et al. (2010) Isolation and characterization of microparticles in sputum of cystic fibrosis patients. Respir Res 11: 94–101.
[29]
McVey MJ, Tabuchi A, Kuebler WM (2012) Microparticles and Acute Lung Injury. Am J Physiol Lung Cell Mol Physiol 303: L364–L381.
[30]
Owens APr, Mackman N (2011) Microparticles in hemostasis and thrombosis. Circ Res 108: 1284–1297.
[31]
Nieuwland R, Berckmans RJ, Rotteveel-Eijkman RC, Maquelin KN, Roozendaal KJ, et al. (1997) Cell-derived microparticles generated in patients during cardiopulmonary bypass are highly procoagulant. Circulation 96: 3534–3541.
[32]
Mallat Z, Benamer H, Hugel B, Benessiano J, Steg PG, et al. (2000) Elevated Levels of Shed Membrane Microparticles With Procoagulant Potential in the Peripheral Circulating Blood of Patients With Acute Coronary Syndromes. Circulation 101: 841–843.
[33]
Preston RA, Jy W, Jimenez JJ, Mauro LM, Horstman LL, et al. (2003) Effects of severe hypertension on endothelial and platelet microparticles. Hypertension 41: 211–217.
[34]
Jung C, Sorensson P, Saleh N, Arheden H, Ryden L, et al. (2011) Circulating endothelial and platelet derived microparticles reflect the size of myocardium at risk in patients with ST-elevation myocardial infarction. Atherosclerosis 221: 226–231.
[35]
Bernimoulin M, Waters EK, Foy M, Steele BM, Sullivan M, et al. (2009) Differential stimulation of monocytic cells results in distinct populations of microparticles. J Thromb Haemost 7: 1019–1028.
[36]
Demedts M, Behr J, Buhl R, Costabel U, Dekhuijzen R, et al. (2005) High-dose acetylcysteine in idiopathic pulmonary fibrosis. N Engl J Med 353: 2229–2242.
[37]
Jose RJ, Williams AE, Chambers RC (2014) Proteinase-activated receptors in fibroproliferative lung disease. Thorax 69: 190–192.
[38]
Curtis AM, Wilkinson PF, Gui M, Gales TL, Hu E, et al. (2009) p38 MAPK targets the production of pro-inflammatory endothelial microparticles. J Thromb Haemost 7: 701–709.
