Genome-Wide Expression Analysis Suggests Hypoxia-Triggered Hyper-Coagulation Leading to Venous Thrombosis at High Altitude

Prabhash Kumar Jha; Anita Sahu; Amit Prabhakar; Tarun Tyagi; Tathagata Chatterjee; Prathima Arvind; Jiny Nair; Neha Gupta; Babita Kumari; Velu Nair; Nitin Bajaj; Jayashree Shanker; Manish Sharma; Bhuvnesh Kumar; Mohammad Zahid Ashraf

doi:10.1055/s-0038-1657770

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2018; 118(07): 1279-1295
DOI: 10.1055/s-0038-1657770

Stroke, Systemic or Venous Thromboembolism

Georg Thieme Verlag KG Stuttgart · New York

Genome-Wide Expression Analysis Suggests Hypoxia-Triggered Hyper-Coagulation Leading to Venous Thrombosis at High Altitude

Prabhash Kumar Jha^*

¹Defense Institute of Physiology and Allied Sciences, Defense Research and Development Organisation, Delhi, India

,

Anita Sahu^*

¹Defense Institute of Physiology and Allied Sciences, Defense Research and Development Organisation, Delhi, India

,

Amit Prabhakar

¹Defense Institute of Physiology and Allied Sciences, Defense Research and Development Organisation, Delhi, India

,

Tarun Tyagi

¹Defense Institute of Physiology and Allied Sciences, Defense Research and Development Organisation, Delhi, India

,

Tathagata Chatterjee

²Army Hospital (Research and Referral), New Delhi, India

,

Prathima Arvind

³Thrombosis Research Institute, India, Bangalore, Karnataka, India

,

Jiny Nair

³Thrombosis Research Institute, India, Bangalore, Karnataka, India

,

Neha Gupta

¹Defense Institute of Physiology and Allied Sciences, Defense Research and Development Organisation, Delhi, India

,

Babita Kumari

¹Defense Institute of Physiology and Allied Sciences, Defense Research and Development Organisation, Delhi, India

,

Velu Nair

⁴Armed Forces Medical College, Pune, Maharashtra, India

,

Nitin Bajaj

⁵Command Hospital (Western Command) Chandimandir, Panchkula, Haryana, India

,

Jayashree Shanker

³Thrombosis Research Institute, India, Bangalore, Karnataka, India

,

Manish Sharma

¹Defense Institute of Physiology and Allied Sciences, Defense Research and Development Organisation, Delhi, India

,

Bhuvnesh Kumar

¹Defense Institute of Physiology and Allied Sciences, Defense Research and Development Organisation, Delhi, India

,

Mohammad Zahid Ashraf^#

¹Defense Institute of Physiology and Allied Sciences, Defense Research and Development Organisation, Delhi, India

› Author Affiliations

Abstract

Venous thromboembolism (VTE), a multi-factorial disease, is the third most common cardiovascular disease. Established genetic and acquired risk factors are responsible for the onset of VTE. High altitude (HA) also poses as an additional risk factor, predisposing individuals to VTE; however, its molecular mechanism remains elusive. This study aimed to identify genes/pathways associated with the pathophysiology of deep vein thrombosis (DVT) at HA. Gene expression profiling of DVT patients, who developed the disease, either at sea level or at HA-DVT locations, resulted in differential expression of 378 and 875 genes, respectively. Gene expression profiles were subjected to bioinformatic analysis, followed by technical and biological validation of selected genes using quantitative reverse transcription-polymerase chain reaction. Both gene ontology and pathway analysis showed enrichment of genes involved in haemostasis and platelet activation in HA-DVT patients with the most relevant pathway being ‘response to hypoxia’. Thus, given the environmental condition the differential expression of hypoxia-responsive genes (angiogenin, ribonuclease, RNase A family, 5; early growth response 1; lamin A; matrix metallopeptidase 14 [membrane-inserted]; neurofibromin 1; PDZ and LIM domain 1; procollagen-lysine 1, 2-oxoglutarate 5-dioxygenase 1; solute carrier family 6 [neurotransmitter transporter, serotonin], member 4; solute carrier family 9 [sodium/hydrogen exchanger], member 1; and TEK tyrosine kinase, endothelial) in HA-DVT could be a determining factor to understand the pathophysiology of DVT at HA.

Keywords

high altitude - microarray - hypoxia - deep vein thrombosis

Full Text

References

References
1 Reitsma PH, Versteeg HH, Middeldorp S. Mechanistic view of risk factors for venous thromboembolism. Arterioscler Thromb Vasc Biol 2012; 32 (03) 563-568
2 Heit JA. Epidemiology of venous thromboembolism. Nat Rev Cardiol 2015; 12 (08) 464-474
3 Moore LG, Niermeyer S, Zamudio S. Human adaptation to high altitude: regional and life-cycle perspectives. Am J Phys Anthropol 1998; 107 (Suppl. 27) 25-64
4 Bendz B, Rostrup M, Sevre K, Andersen TO, Sandset PM. Association between acute hypobaric hypoxia and activation of coagulation in human beings. Lancet 2000; 356 (9242): 1657-1658
5 Hultgren HN. High altitude pulmonary edema. In: High Altitude Medicine. Stanford, CA: Hultgren Publications; 1997
6 Hackett PH, Roach RC. High-altitude medicine. In: Auerbach PS. , ed. Wilderness Medicine. 4th ed. St. Louis, MO: Mosby; 2001
7 Bärtsch P, Schmidt EK, Straub PW. Fibrinopeptide A after strenuous physical exercise at high altitude. J Appl Physiol 1982; 53 (01) 40-43
8 Andrew M, O'Brodovich H, Sutton J. Operation Everest II: coagulation system during prolonged decompression to 282 Torr. J Appl Physiol (1985) 1987; 63 (03) 1262-1267
9 Bärtsch P, Haeberli A, Franciolli M, Kruithof EK, Straub PW. Coagulation and fibrinolysis in acute mountain sickness and beginning pulmonary edema. J Appl Physiol (1985) 1989; 66 (05) 2136-2144
10 Le Roux G, Larmignat P, Marchal M, Richalet JP. Haemostasis at high altitude. Int J Sports Med 1992; 13 (Suppl. 01) S49-S51
11 Hudson JG, Bowen AL, Navia P. , et al. The effect of high altitude on platelet counts, thrombopoietin and erythropoietin levels in young Bolivian airmen visiting the Andes. Int J Biometeorol 1999; 43 (02) 85-90
12 Mannucci PM, Gringeri A, Peyvandi F, Di Paolantonio T, Mariani G. Short-term exposure to high altitude causes coagulation activation and inhibits fibrinolysis. Thromb Haemost 2002; 87 (02) 342-343
13 Tyagi T, Ahmad S, Gupta N. , et al. Altered expression of platelet proteins and calpain activity mediate hypoxia-induced prothrombotic phenotype. Blood 2014; 123 (08) 1250-1260
14 Bradford A. The role of hypoxia and platelets in air travel-related venous thromboembolism. Curr Pharm Des 2007; 13 (26) 2668-2672
15 Kuipers S, Cannegieter SC, Middeldorp S, Robyn L, Büller HR, Rosendaal FR. The absolute risk of venous thrombosis after air travel: a cohort study of 8,755 employees of international organisations. PLoS Med 2007; 4 (09) e290
16 Brill A, Suidan GL, Wagner DD. Hypoxia, such as encountered at high altitude, promotes deep vein thrombosis in mice. J Thromb Haemost 2013; 11 (09) 1773-1775
17 Hiltunen MO, Tuomisto TT, Niemi M. , et al. Changes in gene expression in atherosclerotic plaques analyzed using DNA array. Atherosclerosis 2002; 165 (01) 23-32
18 Chen YC, Bui AV, Diesch J. , et al. A novel mouse model of atherosclerotic plaque instability for drug testing and mechanistic/therapeutic discoveries using gene and microRNA expression profiling. Circ Res 2013; 113 (03) 252-265
19 Koizumi G, Kumai T, Egawa S. , et al. Gene expression in the vascular wall of the aortic arch in spontaneously hypertensive hyperlipidemic model rats using DNA microarray analysis. Life Sci 2013; 93 (15) 495-502
20 Gertow K, Nobili E, Folkersen L. , et al. 12- and 15-lipoxygenases in human carotid atherosclerotic lesions: associations with cerebrovascular symptoms. Atherosclerosis 2011; 215 (02) 411-416
21 King JY, Ferrara R, Tabibiazar R. , et al. Pathway analysis of coronary atherosclerosis. Physiol Genomics 2005; 23 (01) 103-118
22 Levula M, Oksala N, Airla N. , et al. Genes involved in systemic and arterial bed dependent atherosclerosis--Tampere Vascular study. PLoS One 2012; 7 (04) e33787
23 Chohan IS. Blood coagulation changes at high altitude. Def Sci J 1984; 34: 361-379
24 Pichler Hefti J, Risch L, Hefti U. , et al. Changes of coagulation parameters during high altitude expedition. Swiss Med Wkly 2010; 140 (7-8): 111-117
25 Singh I, Chohan IS. Abnormalities of blood coagulation at high altitude. Int J Biometeorol 1972; 16 (03) 283-297
26 Kumar S. High altitude induced deep venous thrombosis: a study of 28 cases. Indian J Surg 2006; 68: 84-88
27 Golebiewska EM, Poole AW. Platelet secretion: from haemostasis to wound healing and beyond. Blood Rev 2015; 29 (03) 153-162
28 Laurens N, Koolwijk P, de Maat MP. Fibrin structure and wound healing. J Thromb Haemost 2006; 4 (05) 932-939
29 Shannon P, Markiel A, Ozier O. , et al. Cytoscape: a software environment for integrated models of biomolecular interaction networks. Genome Res 2003; 13 (11) 2498-2504
30 Huang W, Sherman BT, Lempicki RA. Systematic and integrative analysis of large gene lists using DAVID bioinformatics resources. Nat Protoc 2009; 4 (01) 44-57
31 Franchini M, Lippi G. Von Willebrand factor and thrombosis. Ann Hematol 2006; 85 (07) 415-423
32 Reininger AJ. Function of von Willebrand factor in haemostasis and thrombosis. Haemophilia 2008; 14 (Suppl. 05) 11-26
33 Ruggeri ZM. The role of von Willebrand factor in thrombus formation. Thromb Res 2007; 120 (Suppl. 01) S5-S9
34 Dahm A, Van Hylckama Vlieg A, Bendz B, Rosendaal F, Bertina RM, Sandset PM. Low levels of tissue factor pathway inhibitor (TFPI) increase the risk of venous thrombosis. Blood 2003; 101 (11) 4387-4392
35 Esmon CT. The protein C pathway. Chest 2003; 124 (3, Suppl): 26S-32S
36 Réti M, Farkas P, Csuka D. , et al. Complement activation in thrombotic thrombocytopenic purpura. J Thromb Haemost 2012; 10 (05) 791-798
37 Foley JH, Walton BL, Aleman MM. , et al. Complement activation in arterial and venous thrombosis is mediated by plasmin. EBioMedicine 2016; 5: 175-182
38 Li P, Huang J, Tian HJ, Huang QY, Jiang CH, Gao YQ. Regulation of bone marrow hematopoietic stem cell is involved in high-altitude erythrocytosis. Exp Hematol 2011; 39 (01) 37-46
39 Zovein AC, Forsberg EC. Hematopoietic development at high altitude: blood stem cells put to the test. Development 2015; 142 (10) 1728-1732
40 Reeves JT. Is increased hematopoiesis needed at altitude?. J Appl Physiol (1985) 2004; 96 (05) 1579-1580
41 Hirota K. Involvement of hypoxia-inducible factors in the dysregulation of oxygen homeostasis in sepsis. Cardiovasc Hematol Disord Drug Targets 2015; 15 (01) 29-40
42 Schreiber TD, Steinl C, Essl M. , et al. The integrin alpha9beta1 on hematopoietic stem and progenitor cells: involvement in cell adhesion, proliferation and differentiation. Haematologica 2009; 94 (11) 1493-1501
43 Cambi A, van Helden SFG, Figdor CG. Roles for integrins and associated proteins in the haematopoietic system. In: Madame Curie Bioscience Database. Austin, TX: Landes Bioscience; 2000-2013
44 Goggins BJ, Chaney C, Radford-Smith GL, Horvat JC, Keely S. Hypoxia and integrin-mediated epithelial restitution during mucosal inflammation. Front Immunol 2013; 4: 272
45 Bergmeier W, Hynes RO. Extracellular matrix proteins in hemostasis and thrombosis. Cold Spring Harb Perspect Biol 2012; 4 (02) a005132
46 Nieswandt B, Watson SP. Platelet-collagen interaction: is GPVI the central receptor?. Blood 2003; 102 (02) 449-461
47 Croft D, Mundo AF, Haw R. , et al. The Reactome pathway knowledgebase. Nucleic Acids Res 2014; 42 (Database issue): D472-D477
48 Bindea G, Mlecnik B, Hackl H. , et al. ClueGO: a Cytoscape plug-in to decipher functionally grouped gene ontology and pathway annotation networks. Bioinformatics 2009; 25 (08) 1091-1093
49 Bärtsch P, Waber U, Haeberli A. , et al. Enhanced fibrin formation in high-altitude pulmonary edema. J Appl Physiol (1985) 1987; 63 (02) 752-757
50 Vij AG. Effect of prolonged stay at high altitude on platelet aggregation and fibrinogen levels. Platelets 2009; 20 (06) 421-427
51 Lehmann T, Mairbäurl H, Pleisch B, Maggiorini M, Bärtsch P, Reinhart WH. Platelet count and function at high altitude and in high-altitude pulmonary edema. J Appl Physiol (1985) 2006; 100 (02) 690-694
52 Rahaman MM, Reinders FG, Koes D. , et al. Structure guided chemical modifications of propylthiouracil reveal novel small molecule inhibitors of cytochrome b5 reductase 3 that increase nitric oxide bioavailability. J Biol Chem 2015; 290 (27) 16861-16872
53 Freedman JE, Loscalzo J. Nitric oxide and its relationship to thrombotic disorders. J Thromb Haemost 2003; 1 (06) 1183-1188
54 Hull CM, Rajendran D, Fernandez Barnes A. Deep vein thrombosis and pulmonary embolism in a mountain guide: awareness, diagnostic challenges, and management considerations at altitude. Wilderness Environ Med 2016; 27 (01) 100-106
55 Gupta N, Sahu A, Prabhakar A. , et al. Activation of NLRP3 inflammasome complex potentiates venous thrombosis in response to hypoxia. Proc Natl Acad Sci U S A 2017; 114 (18) 4763-4768
56 Bovill EG, van der Vliet A. Venous valvular stasis-associated hypoxia and thrombosis: what is the link?. Annu Rev Physiol 2011; 73: 527-545
57 Yan SF, Mackman N, Kisiel W, Stern DM, Pinsky DJ. Hypoxia/hypoxemia-induced activation of the procoagulant pathways and the pathogenesis of ischemia-associated thrombosis. Arterioscler Thromb Vasc Biol 1999; 19 (09) 2029-2035
58 Naukkarinen J, Gentile M, Soro-Paavonen A. , et al. USF1 and dyslipidemias: converging evidence for a functional intronic variant. Hum Mol Genet 2005; 14 (17) 2595-2605
59 Fan Y-M, Hernesniemi J, Oksala N. , et al. Upstream Transcription Factor 1 (USF1) allelic variants regulate lipoprotein metabolism in women and USF1 expression in atherosclerotic plaque. Sci Rep 2014; 4 (04) 4650
60 Willam C, Koehne P, Jürgensen JS. , et al. Tie2 receptor expression is stimulated by hypoxia and proinflammatory cytokines in human endothelial cells. Circ Res 2000; 87 (05) 370-377
61 Kishimoto K, Yoshida S, Ibaragi S. , et al. Hypoxia-induced up-regulation of angiogenin, besides VEGF, is related to progression of oral cancer. Oral Oncol 2012; 48 (11) 1120-1127
62 Land SC, Tee AR. Hypoxia-inducible factor 1α is regulated by the mammalian target of rapamycin (mTOR) via an mTOR signaling motif. J Biol Chem 2007; 282 (28) 20534-20543
63 Gilkes DM, Bajpai S, Wong CC. , et al. Procollagen lysyl hydroxylase 2 is essential for hypoxia-induced breast cancer metastasis. Mol Cancer Res 2013; 11 (05) 456-466
64 Tan PK, Downey TJ, Spitznagel Jr EL. , et al. Evaluation of gene expression measurements from commercial microarray platforms. Nucleic Acids Res 2003; 31 (19) 5676-5684
65 Leek JT, Johnson WE, Parker HS, Jaffe AE, Storey JD. The SVA package for removing batch effects and other unwanted variation in high-throughput experiments. Bioinformatics 2012; 28 (06) 882-883
66 Parry RM, Jones W, Stokes TH. , et al. k-Nearest neighbor models for microarray gene expression analysis and clinical outcome prediction. Pharmacogenomics J 2010; 10 (04) 292-309
67 Chu L, Scharf E, Kondo T. GeneSpring: tools for analyzing microarray expression data. Genome Informatics 2001; 12: 227-229
68 Maere S, Heymans K, Kuiper M. BiNGO: a Cytoscape plugin to assess overrepresentation of gene ontology categories in biological networks. Bioinformatics 2005; 21 (16) 3448-3449
69 Wu G, Dawson E, Duong A. , et al. ReactomeFIViz: the Reactome FI Cytoscape app for pathway and network-based data analysis. [version 1; referees: 1 approved, 1 approved with reservations] F1000 Res 2014
70 Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods 2001; 25 (04) 402-408

Supplementary Material