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Orphanet Journal of Rare Diseases

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Open Access 01-12-2015 | Research

Molecular and clinical analysis of TRPC6 and AGTR1 genes in patients with pulmonary arterial hypertension

Authors: Guillermo Pousada, Adolfo Baloira, Diana Valverde

Published in: Orphanet Journal of Rare Diseases | Issue 1/2015

Abstract

Background

Pulmonary arterial hypertension (PAH) is a rare and progressive vascular disorder characterized by increased pulmonary vascular resistance and right heart failure. The aim of this study was to analyze 5′UTR region in canonical transient receptor potential isoform 6 (TRPC6) and 3′UTR region in Angiotensin II type I receptor (AGTR1) genes in patients with idiopathic and associated PAH. Correlation among mutations and clinical and functional parameters was further analyzed.

Methods

Analysis of TRPC6 and AGTR1 genes was performed by polymerase chain reaction (PCR) and direct sequencing. We used a non-parametric test to determine if significant differences were found between the groups studied and chi-square test to compare clinical and hemodynamic variables among genotypes.

Results

Fifty five patients and fifty two controls were included in this study. We found statistically significant differences for c.1-361A > T (p = 0.0077), c.1-254C > G (p < 0.0001) and c.1-218C > T (p = 0.0021) in TRPC6 gene and c.1166A > C (p < 0.001) in AGTR1 gene, between patients and controls. Idiopathic PAH patients (IPAH) and controls presented significant differences for all 3 TRPC6 polymorphisms (p = 0.020), (p = 0.002) and (p = 0.008) respectively, and also showed differences for AGTR1 gene (p < 0.001). In associated PAH (APAH) patients we found statistical differences for c.1-254C > G (p < 0.001) and c.1-218C > T (p = 0.001) in TRPC6 gene and c.1166A > C (p = 0.001) in AGTR1 gene. Several clinical and hemodynamic parameters showed significant differences between carriers and non-carriers of these single nucleotide polymorphisms (SNPs). Nineteen patients were carriers of all 3 SNPs in TRPC6 gene and presented a more severe phenotype with differences in mean pulmonary arterial pressure (p = 0.016), systolic pulmonary arterial pressure (p = 0.040), cardiac index (p < 0.001) and 6 minute walking test (p = 0.049). 16 of these patients harbored the SNP in AGTR1 gene. These patients showed differences in age at diagnosis (p = 0.049), mean pulmonary arterial pressure (p = 0.033), cardiac index (p = 0.002) and 6 minute walking test (p = 0.039).

Conclusions

PAH is a rare disease with pulmonary vascular remodeling caused in part by a heterogeneous constellation of genetic arrangements. This study seems to suggest that c.1-361A > T, c.1-254C > G and c.1-218C > T polymorphisms in TRPC6 gene and c.1166A > C polymorphism in AGTR1 could have a role in the development of this disease.

Hoeper MM, Bogaard HJ, Condliffe R, Frantz R, Khanna D, Kurzyna M, et al. Definitions and diagnosis of pulmonary hypertension. J Am Coll Cardiol. 2013;62:25S.CrossRef

Peacock AJ, Murphy NF, McMurray JJV, Caballero L, Stewart S. An epidemiological study of pulmonary arterial hypertension. Eur Respir J. 2007;30:104–9.PubMedCrossRef

Sanchez O, Marié E, Lerolle U, Wermert D, Israël-Biel D, Meyer G. Pulmonary arterial hypertension in women. Rev Mal Respir. 2010;27:e79–87.PubMedCrossRef

D’Alonzo GE, Barst RJ, Ayres SM, Bergofsky EH, Brundage BH, Detre KM, et al. Survival in patients with primary pulmonary hypertension. Results from a national prospective registry. Ann Intern Med. 1991;115:343–9.PubMedCrossRef

Simonneau G, Gatzoulis MA, Adatia I, Celermajer D, Denton C, Ghofrani A, et al. Updated clinical classification of pulmonary hypertension. J Am Coll Cardiol. 2013;62:25s.CrossRef

Soubrier F, Chung WK, Machado R, Grünig E, Aldred M, Geraci M, et al. Genetics and genomics of pulmonary arterial hypertension. J Am Coll Cardiol. 2013;62:25S.CrossRef

Baloira A, Vilariño C, Leiro V, Valverde D. Mutations in the gene encoding bone morphogenetic protein receptor 2 in patients with idiopathic pulmonary arterial hypertension. Arch Bronconeumol. 2008;44(1):29–34.PubMedCrossRef

Machado RD, Eickelberg O, Elliott G, Geraci MW, Hanaoka M, Loyd JE, et al. Genetics and genomics of pulmonary arterial hypertension. J Am Coll Cardiol. 2009;54 Suppl 1:S32–42.PubMedCentralPubMedCrossRef

Yang X, Long L, Reynolds PN, Morrell NW. Expression of mutant BMPR-II in pulmonary endothelial cells promotes apoptosis and a release of factors that stimulate proliferation of pulmonary arterial smooth muscle cells. Pulm Circ. 2011;1(1):103–11.PubMedCentralPubMedCrossRef

10.

Pffar N, Szamalek-Hoegel J, Fischer C, Hinderhofer K, Nagel C, Eklken N, et al. Hemodynamic and clinical onset in patients with hereditary pulmonary arterial hypertension and BMPR2 mutations. Respir Res. 2011;12:99–109.CrossRef

11.

Ulrich S, Szamalek-Hoegel J, Hersberger M, Fischler M, García JS, Huber LC, et al. Sequence variants in BMPR2 and genes involved in the serotonin and nitric oxide pathways in idiopathic pulmonary arterial hypertension and chronic thromboembolic pulmonary hypertension: relation to clinical parameters and comparison with left heart disease. Respiration. 2010;79:279–87.PubMedCrossRef

12.

Pousada G, Baloira A, Vilariño C, Cifrián JM, Valverde D. Novel mutations in BMPR2, ACVRL1 and KCNA5 genes and hemodynamic parameters in patients with pulmonary arterial hypertension. PLoS ONE. 2014;9(6):e100261.PubMedCentralPubMedCrossRef

13.

Pfarr N, Fischer C, Ehlken N, Becker-Grünig T, López-González V, Gorenflo M, et al. Hemodynamic and genetic analysis in children with idiopathic, heritable, and congenital heart disease associated pulmonary arterial hypertension. Respir Res. 2013;14:3–12.PubMedCentralPubMedCrossRef

14.

Yu Y, Fantozzi I, Remillard CV, Landsberg JW, Kunichika N, Plastoshyn O, et al. Enhanced expression of transient receptor potential channels in idiopathic pulmonary arterial hypertension. PNAS. 2004;101(38):13861–6.PubMedCentralPubMedCrossRef

15.

Xia Y, Yang XR, Fu Z, Paudel O, Abramowitz J, Birnbaumer L, et al. Classical transient receptor potential 1 and 6 contribute to hypoxic pulmonary hypertension through differential regulation of pulmonary vascular functions. Hypertension. 2014;63:173–80.PubMedCentralPubMedCrossRef

16.

Alzoubi A, Almalouf P, Toba M, O’Neill K, Qian X, Francis M, et al. TRPC4 inactivation confers a survival benefit in severe pulmonary arterial hypertension. Am J Pathol. 2013;183:1779–88.PubMedCrossRef

17.

Weissmann N, Dietrich A, Fuchs B, Kalwa H, Ay M, Dumitrascu R, et al. Classical transient receptor potential channel 6 (TRPC6) is essential for hypoxic pulmonary vasoconstriction and alveolar gas exchange. Proc Natl Acad Sci U S A. 2006;103:19093–8.PubMedCentralPubMedCrossRef

18.

Zhang Y, Lu W, Yang K, Xu L, Lai N, Tian L, et al. Bone morphogenetic protein 2 decreases TRPC expression, store-operated Ca(2+) entry, and basal [Ca(2+)]i in rat distal pulmonary arterial smooth muscle cells. Am J Physiol Cell Physiol. 2013;304:C833–43.PubMedCentralPubMedCrossRef

19.

Yu Y, Keller SH, Remillard CV, Safrina O, Nicholson A, Zhang SL, et al. A functional single-nucleotide polymorphism in the TRPC6 gene promoter associated with idiopathic pulmonary arterial hypertension. Circulation. 2009;119:2313–22.PubMedCentralPubMedCrossRef

20.

Palatini P, Ceolotto G, Dorigatti F, Mos L, Santonastaso M, Bratti P, et al. Angiotensin II type 1 receptor gene polymorphism predicts development of hypertension and metabolic syndrome. Am J Hypertens. 2009;22:208–14.PubMedCrossRef

21.

Nie SJ, Wen-ru T, Bi-feng C, Jin L, Wen Z, Sheng-jun L, et al. Haplotype-based case–control study of the human AGTR1 gene and essential hypertension in Han Chinese subjects. Clin Biochem. 2010;43(3):253–8.PubMedCrossRef

22.

Chassagne C, Eddahibi S, Adamy C, Rideau D, Marotte F, Dubois-Randé J, et al. Modulation of angiotensin II receptor expression during development and regression of hypoxic pulmonary hypertension. Am J Respir Cell Mol Biol. 2000;22:323–32.PubMedCrossRef

23.

Li Y, Li X, Jia N, Guo S, Chu S, Niu W. Meta-analysis of the association between angiotensin II receptor, type 1 gene A1166C polymorphism and coronary artery disease in Chinese populations. J Renin Angiotensin Aldosterone Syst. 2013;14(1):82–90.PubMedCrossRef

24.

Gardier S, Vincent M, Lantelme P, Rial MO, Bricca G, Milon H. A1166C polymorphism of angiotensin II type 1 receptor, blood pressure and arterial stiffness in hypertension. J Hypertens. 2004;22:2135–42.PubMedCrossRef

25.

Chung WK, Deng L, Carroll JS, Mallory N, Diamond B, Rosenzweig EB, et al. Polymorphism in the angiotensin II type 1 receptor (AGTR1) is associated with age at diagnosis in pulmonary arterial hypertension. J Heart Lung Transplant. 2009;28:373–9.PubMedCentralPubMedCrossRef

26.

Galié N, Hoeper MM, Humbert M, Torbicki A, Vachiery JL, Barbera JA, et al. Guidelines for the diagnosis and treatment of pulmonary hypertension. Eur Heart J. 2009;30(20):2493–537.PubMedCrossRef

27.

Jíra M, Závodná E, Honzíková N, Nováková Z, Vasků A, Izakovicová Hollá L, et al. Association of A1166C polymorphism in AT(1) receptor gene with baroreflex sensitivity. Physiol Res. 2010;59(4):517–28.PubMed

28.

Inoue R, Jensen LJ, SHI J, Morita H, Nishida M, Honda A, et al. Transient receptor potential channels in cardiovascular function and disease. Circ Res. 2006;99:119–31.PubMedCrossRef

29.

Freichel M, Vennekens R, Olausson J, Stolz S, Philipp SE, WeiBgerber P, et al. Functional role of TRPC proteins in native systems: implications from knockout and knock-down studies. J Physiol. 2005;567.1:59–66.CrossRef

30.

Lu W, Ran P, Zhang D, Lai N, Zhong N, Wang J. Bone morphogenetic protein 4 enhances canonical transient receptor potential expression, store-operated Ca₂ ⁺ entry, and basal [Ca₂]_i in rat distal pulmonary arterial smooth muscle cells. Am J Physiol Cell Physiol. 2010;299:C1370–8.PubMedCentralPubMedCrossRef

31.

Kunichika N, Landsberg JW, Yu Y, Kunichika H, Thistlethwaite PA, Rubin LJ, et al. Bosentan inhibits transient receptor potential channel expression in pulmonary vascular myocytes. Am J Respir Crit Care Med. 2004;170:1101–7.PubMedCrossRef

32.

Ogawa A, Firth AL, Yao W, Rubin LJ, Yuan JX-J. Prednisolone inhibits PDGF-induced nuclear translocation of NF-κB in human pulmonary artery smooth muscle cells. Am J Physiol Lung Cell Mol Physiol. 2008;295:L648–57.PubMedCentralPubMedCrossRef

33.

Boney-Montoya J, Ziegler YS, Curtis CD, Montoya JA, Nardulli AM. Long-range transcriptional control of progesterone receptor gene expression. Mol Endocrinol. 2010;24(2):346–58.PubMedCentralPubMedCrossRef

34.

Xu M, Sham P, Ye Z, Lindpaintner K, He L. A 1166C genetic variation of the angiotensin II type I receptor gene and susceptibility to coronary heart disease: collaborative of 53 studies with 20,435 cases and 23,674 controls. Atherosclerosis. 2010;213(1):30–2.CrossRef

35.

Sethupathy P, Borel C, Gagnebin M, Grant GR, Deutsch S, Elton TS, et al. Human microRNA-155 on chromosome 21 differentially interacts with its polymorphic target in the AGTR1 3′ untranslated region: a mechanism for functional single-nucleotide polymorphisms related to phenotypes. Am J Hum Genet. 2007;81(2):405–13.PubMedCentralPubMedCrossRef

36.

Haas U, Sczakiel G, Laufer SD. MicroRNA-mediated regulation of gene expression is affected by disease-associated SNPs within the 3′-UTR via altered RNA structure. RNA Biol. 2012;9(6):924–37.PubMedCentralPubMedCrossRef

37.

Song S, Yamamura A, Yamamura H, Ayon RJ, Smith KA, Tang H, et al. Flow shear stress enhances intracellular Ca²⁺ signaling in pulmonary artery smooth muscle cells from patients with pulmonary arterial hypertension. Am J Physiol Cell Physiol. 2014;307(4):C373–83.PubMedCrossRef

Title: Molecular and clinical analysis of TRPC6 and AGTR1 genes in patients with pulmonary arterial hypertension
Authors: Guillermo Pousada
Adolfo Baloira
Diana Valverde
Publication date: 01-12-2015
Publisher: BioMed Central
Published in: Orphanet Journal of Rare Diseases / Issue 1/2015
Electronic ISSN: 1750-1172
DOI: https://doi.org/10.1186/s13023-014-0216-3

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Molecular and clinical analysis of TRPC6 and AGTR1 genes in patients with pulmonary arterial hypertension

Abstract

Background

Methods

Results

Conclusions

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Background

Methods

Results

Conclusions

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