Characterization of proximal pulmonary arterial cells from chronic thromboembolic pulmonary hypertension patients

Respiratory Research - Tập 13 Số 1 - 2012
Rozenn Quarck1, Marijke Wynants1, Alicja Ronisz1, M. Rosario Sepúlveda2, Frank Wuytack2, Dirk Van Raemdonck3, Bart Meyns4, Marion Delcroix5
1Center for Pulmonary Vascular Diseases, Respiratory Disease Department, Katholieke Universiteit and Universitaire Ziekenhuizen Leuven, Leuven, Belgium
2Molecular Cell Biology Department, Katholieke Universiteit, Leuven, Belgium
3Thoracic Surgery and Cardiac Surgery Department, Universitaire Ziekenhuizen Leuven, Leuven, Belgium
4Cardiac Surgery Department, Universitaire Ziekenhuizen Leuven, Leuven, Belgium
5Department of Pneumology, Universitaire Ziekenhuizen Leuven, Herestraat 49, B-3000, Leuven, Belgium

Tóm tắt

Abstract Background Chronic thromboembolic pulmonary hypertension (CTEPH) is associated with proximal pulmonary artery obstruction and vascular remodeling. We hypothesized that pulmonary arterial smooth muscle (PASMC) and endothelial cells (PAEC) may actively contribute to remodeling of the proximal pulmonary vascular wall in CTEPH. Our present objective was to characterize PASMC and PAEC from large arteries of CTEPH patients and investigate their potential involvement in vascular remodeling. Methods Primary cultures of proximal PAEC and PASMC from patients with CTEPH, with non-thromboembolic pulmonary hypertension (PH) and lung donors have been established. PAEC and PASMC have been characterized by immunofluorescence using specific markers. Expression of smooth muscle specific markers within the pulmonary vascular wall has been studied by immunofluorescence and Western blotting. Mitogenic activity and migratory capacity of PASMC and PAEC have been investigated in vitro. Results PAEC express CD31 on their surface, von Willebrand factor in Weibel-Palade bodies and take up acetylated LDL. PASMC express various differentiation markers including α-smooth muscle actin (α-SMA), desmin and smooth muscle myosin heavy chain (SMMHC). In vascular tissue from CTEPH and non-thromboembolic PH patients, expression of α-SMA and desmin is down-regulated compared to lung donors; desmin expression is also down-regulated in vascular tissue from CTEPH compared to non-thromboembolic PH patients. A low proportion of α-SMA positive cells express desmin and SMMHC in the neointima of proximal pulmonary arteries from CTEPH patients. Serum-induced mitogenic activity of PAEC and PASMC, as well as migratory capacity of PASMC, were increased in CTEPH only. Conclusions Modified proliferative and/or migratory responses of PASMC and PAEC in vitro, associated to a proliferative phenotype of PASMC suggest that PASMC and PAEC could contribute to proximal vascular remodeling in CTEPH.

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Tài liệu tham khảo

Hoeper MM, Mayer E, Simonneau G, Rubin LJ: Chronic thromboembolic pulmonary hypertension. Circulation. 2006, 113: 2011-2020. 10.1161/CIRCULATIONAHA.105.602565.

Arbustini E, Morbini P, D'Armini AM, Repetto A, Minzioni G, Piovella F, Vigano M, Tavazzi L: Plaque composition in plexogenic and thromboembolic pulmonary hypertension: the critical role of thrombotic material in pultaceous core formation. Heart. 2002, 88: 177-182. 10.1136/heart.88.2.177.

Bonderman D, Turecek PL, Jakowitsch J, Weltermann A, Adlbrecht C, Schneider B, Kneussl M, Rubin LJ, Kyrle PA, Klepetko W, Maurer G, Lang IM: High prevalence of elevated clotting factor VIII in chronic thromboembolic pulmonary hypertension. Thromb Haemost. 2003, 90: 372-376.

Bonderman D, Wilkens H, Wakounig S, Schafers HJ, Jansa P, Lindner J, Simkova I, Martischnig AM, Dudczak J, Sadushi R, Skoro-Sajer N, Klepetko W, Lang IM: Risk factors for chronic thromboembolic pulmonary hypertension. Eur Resp J. 2009, 33: 325-331.

Lang I, Kerr K: Risk factors for chronic thromboembolic pulmonary hypertension. Proc Am Thorac Soc. 2006, 3: 568-570. 10.1513/pats.200605-108LR.

Bonderman D, Jakowitsch J, Adlbrecht C, Schemper M, Kyrle PA, Schonauer V, Exner M, Klepetko W, Kneussl MP, Maurer G, Lang I: Medical conditions increasing the risk of chronic thromboembolic pulmonary hypertension. Thromb Haemost. 2005, 93: 512-516.

Quarck R, Nawrot T, Meyns B, Delcroix M: C-reactive protein: a new predictor of adverse outcome in pulmonary arterial hypertension. J Am Coll Cardiol. 2009, 53: 1211-1218. 10.1016/j.jacc.2008.12.038.

Bernard J, Yi ES: Pulmonary thromboendarterectomy: a clinicopathologic study of 200 consecutive pulmonary thromboendarterectomy cases in one institution. Hum Pathol. 2007, 38: 871-877. 10.1016/j.humpath.2006.11.017.

Pusztaszeri MP, Seelentag W, Bosman FT: Immunohistochemical Expression of Endothelial Markers CD31, CD34, von Willebrand Factor, and Fli-1 in Normal Human Tissues. J Histochem Cytochem. 2006, 54: 385-395. 10.1369/jhc.4A6514.2005.

Tamura Y, Osuga Ji, Adachi H, Tozawa Ri, Takanezawa Y, Ohashi K, Yahagi N, Sekiya M, Okazaki H, Tomita S, Iizuka Y, Koizumi H, Inaba T, Yagyu H, Kamada N, Suzuki H, Shimano H, Kadowaki T, Tsujimoto M, Arai H, Yamada N, Ishibashi S: Scavenger Receptor Expressed by Endothelial Cells I (SREC-I) Mediates the Uptake of Acetylated Low Density Lipoproteins by Macrophages Stimulated with Lipopolysaccharide. J Biol Chem. 2004, 279: 30938-30944. 10.1074/jbc.M313088200.

Owens GK: Regulation of differentiation of vascular smooth muscle cells. Physiol Rev. 1995, 75: 487-517.

Theilmeier G, Quarck R, Verhamme P, Bochaton-Piallat ML, Lox M, Bernar H, Janssens S, Kockx M, Gabbiani G, Collen D, Holvoet P: Hypercholesterolemia impairs vascular remodelling after porcine coronary angioplasty. Cardiovasc Res. 2002, 55: 385-395. 10.1016/S0008-6363(02)00409-1.

Quarck R, Berrou E, Magnier C, Bobe R, Bredoux R, Tobelem G, Enouf J, Bryckaert M: Differential up-regulation of Rap1a and Rap1b proteins during smooth muscle cell cycle. Eur J Cell Biol. 1996, 70: 269-277.

Sacks RS, Remillard CV, Agange N, Auger WR, Thistlethwaite PA, Yuan JX: Molecular biology of chronic thromboembolic pulmonary hypertension. Semin Thorac Cardiovasc Surg. 2006, 18: 265-276. 10.1053/j.semtcvs.2006.09.004.

Lang IM, Marsh JJ, Olman MA, Moser KM, Schleef RR: Parallel analysis of tissue-type plasminogen activator and type 1 plasminogen activator inhibitor in plasma and endothelial cells derived from patients with chronic pulmonary thromboemboli. Circulation. 1994, 90: 706-712.

Firth AL, Yao W, Ogawa A, Madani MM, Lin GY, Yuan JXJ: Multipotent mesenchymal progenitor cells are present in endarterectomized tissues from patients with chronic thromboembolic pulmonary hypertension. Am J Physiol Cell Physiol. 2010, 298: C1217-C1225. 10.1152/ajpcell.00416.2009.

Yao W, Firth AL, Sacks RS, Ogawa A, Auger WR, Fedullo PF, Madani MM, Lin GY, Sakakibara N, Thistlethwaite PA, Jamieson SW, Rubin LJ, Yuan JXJ: Identification of putative endothelial progenitor cells (CD34 + CD133 + Flk-1+) in endarterectomized tissue of patients with chronic thromboembolic pulmonary hypertension. Am J Physiol Lung Cell Mol Physiol. 2009, 296: L870-L878. 10.1152/ajplung.90413.2008.

Yan Q, Vernon RB, Hendrickson AE, Sage EH: Primary culture and characterization of microvascular endothelial cells from Macaca monkey retina. Investig Ophthalmol Vis Sci. 1996, 37: 2185-2194.

Owens GK, Kumar MS, Wamhoff BR: Molecular regulation of vascular smooth muscle cell differentiation in development and disease. Physiol Rev. 2004, 84: 767-801. 10.1152/physrev.00041.2003.

Dewachter L, Adnot S, Fadel E, Humbert M, Maitre B, Barlier-Mur AM, Simonneau G, Hamon M, Naeije R, Eddahibi S: Angiopoietin/Tie2 Pathway Influences Smooth Muscle Hyperplasia in Idiopathic Pulmonary Hypertension. Am J Resp Crit Care Med. 2006, 174: 1025-1033. 10.1164/rccm.200602-304OC.

Eddahibi S, Guignabert C, Barlier-Mur AM, Dewachter L, Fadel E, Dartevelle P, Humbert M, Simonneau G, Hanoun N, Saurini F, Hamon M, Adnot S: Cross Talk Between Endothelial and Smooth Muscle Cells in Pulmonary Hypertension: Critical Role for Serotonin-Induced Smooth Muscle Hyperplasia. Circulation. 2006, 113: 1857-1864. 10.1161/CIRCULATIONAHA.105.591321.

Masri FA, Xu W, Comhair SAA, Asosingh K, Koo M, Vasanji A, Drazba J, Anand-Apte B, Erzurum SC: Hyperproliferative apoptosis-resistant endothelial cells in idiopathic pulmonary arterial hypertension. Am J Physiol Lung Cell Mol Physiol. 2007, 293: L548-L554. 10.1152/ajplung.00428.2006.

Xu W, Koeck T, Lara AR, Neumann D, DiFilippo FP, Koo M, Janocha AJ, Masri FA, Arroliga AC, Jennings C, Dweik RA, Tuder RM, Stuehr DJ, Erzurum SC: Alterations of cellular bioenergetics in pulmonary artery endothelial cells. Proc Natl Acad Sci USA. 2007, 104: 1342-1347. 10.1073/pnas.0605080104.

Blauwet LA, Edwards WD, Tazelaar HD, McGregor CG: Surgical pathology of pulmonary thromboendarterectomy: a study of 54 cases from 1990 to 2001. Hum Pathol. 2003, 34: 1290-1298. 10.1016/j.humpath.2003.07.003.

Jamieson SW, Kapelanski DP, Sakakibara N, Manecke GR, Thistlethwaite PA, Kerr KM, Channick RN, Fedullo PF, Auger WR: Pulmonary endarterectomy: experience and lessons learned in 1,500 cases. Ann Thorac Surg. 2003, 76: 1457-1462. 10.1016/S0003-4975(03)00828-2.

Yi ES, Kim H, Ahn H, Strother J, Morris T, Masliah E, Hansen LA, Park K, Friedman PJ: Distribution of obstructive intimal lesions and their cellular phenotypes in chronic pulmonary hypertension. A morphometric and immunohistochemical study. Am J Respir Crit Care Med. 2000, 162: 1577-1586.