Status of serine tyrosine kinase at germline and expressional levels in asthma patients

Document Type : Original article

Authors

1 Department of Biosciences, COMSATS Institute of Information Technology, Islamabad, Pakistan

2 Department of Environmental Sciences, Fatima Jinnah Women University, Rawalpindi, Pakistan

3 Department of Microbiology, Faculty of Biological Sciences, Quaid-i-Azam University, Islamabad, Pakistan

Abstract

Asthma is a disease marked by inflammation of airways with an increasing incidence rate worldwide especially among Asian population. Spleen tyrosine kinase (Syk) is known to be involved in regulation of such inflammation response and thereby rendering its inevitable importance among asthma patients. DNA extraction followed by PCR and sequencing was performed for genomic analysis, mRNA analysis was done by RT PCR whereas Western blot and ELISA was used for protein study. Image J and UNAFOLD were also used for Bioinformatics analysis.The mean age of patients and controls were 31.1±9.3 and 30.4±6.1 years respectively. Results of sequencing showed nonsense exonic mutations in exon 3 at g.25710G>A and g.25722G>A positions. Substitution mutations in introns were also found at g.25827G>A (intron 3), g.63425C>T (intron 8) and g.63445T>G (intron 8). Significantly increased levels of IgE and significantly decreased expression of Syk at transcriptional level was found in patients compared to controls. The western blot results of asthmatic samples and healthy controls revealed that Syk has comparatively low expression in diseased individual’s PBMCs.SYK has been found to be altered in DNA, mRNA and protein expression in asthma patients among Pakistani population therefore patients should be treated according to their Syk status for more effective recovery.

Keywords

References

1. Dougherty RH, Fahy JV. Acute exacerbations of asthma: epidemiology, biology and the exacerbation-prone phenotype. Clin Exp Allergy 2009;39:193-202.
2. Passalacqua G, Ciprandi G. Allergy and the lung. Clin Exp Immunol 2008;153 Suppl 1:12-16.
3. Nauta AJ, Engels F, Knippels LM, Garssen J, Nijkamp FP, Redegeld FA. Mechanisms of allergy and asthma. Eur J Pharmacol 2008;585:354-360.
4. Kim HY, DeKruyff RH, Umetsu DT. The many paths to asthma: phenotype shaped by innate and adaptive immunity. Nature Immunol 2010;11:577-584.
5. Gould HJ, Sutton BJ, Beavil AJ, Beavil RL, McCloskey N, Coker HA, Fear D, Smurthwaite L. The biology of IGE and the basis of allergic disease. Ann Rev Immunol 2003;21:579-628.
6. Zhang M, Murphy RF, Agrawal DK. Decoding IgE Fc receptors. Immunologic Res 2007;37: 1-16.
7. Metz M, Grimbaldeston MA, Nakae S, Piliponsky AM, Tsai M, Galli SJ. Mast cells in the promotion and limitation of chronic inflammation. Immunol Rev 2007;217:304-328.
8. Hubbard SR, Till JH. Protein tyrosine kinase structure and function. Ann Rev Biochem 2000; 69:373-398.
9. Latour S, Veillette A. Proximal protein tyrosine kinases in immunoreceptor signaling. Curr Opin Immunol 2001;13:299-306.
10. Muller B, Cooper L, Terhorst C. Molecular cloning of the human homologue to the pig protein-tyrosine kinase syk. Immunogenetics 1994;39:359-362.
11. Siraganian RP, Zhang J, Suzuki K, Sada K. Protein tyrosine kinase Syk in mast cell signaling. Mol Immunol 2002;38:1229-1233.
12. Daeron M. Fc receptor biology. Ann Rev Immunol 1997;15:203-234.
13. Turner M, Schweighoffer E, Colucci F, Di Santo JP, Tybulewicz VL. Tyrosine kinase SYK: essential functions for immunoreceptor signalling. Immunol Today 2000;21:148-154.
14. Wang L, Duke L, Zhang PS, Arlinghaus RB, Symmans WF, Sahin A, Mendez R, Dai JL. Alternative splicing disrupts a nuclear localization signal in spleen tyrosine kinase that is required for invasion suppression in breast cancer. Cancer Res 2003;63:4724-4730.
15. Latour S, Zhang J, Siraganian RP, Veillette A. A unique insert in the linker domain of Syk is necessary for its function in immunoreceptor signalling. EMBO J 1998;17:2584-2595.
16. Ahmed A, Minhas K, Micheal S, Ahmad F. Prevalence of skin test reactivity to aeroallergens in the Pakistani population. Public Health 2011;125:324-326.
17. Nosheen M, Ishrat M, Malik FA, Baig RM, Kayani MA. Association of GSTM1 and GSTT1 gene deletions with risk of head and neck cancer in Pakistan: a case control study. Asian Pac J Cancer Prev 2010;11:881-885.
18. Towbin H, Staehelin T, Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Biotechnology 1992;24:145-149.
19. Nanes BA. Slide Set: Reproducible image analysis and batch processing with Image J. BioTechniques 2015;59:269-278.
20. Coopman PJ, Do MT, Barth M, Bowden ET, Hayes AJ, Basyuk E, Blancato JK, Vezza PR, McLeskey SW, Mangeat PH, Muller SC. The Syk tyrosine kinase suppresses malignant growth of human breast cancer cells. Nature 2000;406:742-747.
21. Matsubara S, Li G, Takeda K, Loader JE, Pine P, Masuda ES, Miyahara N, Miyahara S, Lucas JJ, Dakhama A, Gelfand EW. Inhibition of spleen tyrosine kinase prevents mast cell activation and airway hyperresponsiveness. Am J respiratory critical care Med 2006;173:56-63.
22. Scheijen B, Griffin JD. Tyrosine kinase oncogenes in normal hematopoiesis and hematological disease. Oncogene 2002;21:3314-3333.
23. Hulme JS, Barratt BJ, Twells RC, Cooper JD, Lowe CE, Howson JM, Lam AC, Smink LJ, Savage DA, Undlien DE, Guja C, Ionescu-Tîirgoviste C, Tuomilehto-Wolf E, Tuomilehto J, Todd JA. Association analysis of the lymphocyte-specific protein tyrosine kinase (LCK) gene in type 1 diabetes. Diabetes 2004;53:2479-2482.
24. Schatz M, Camargo CA, Jr. The relationship of sex to asthma prevalence, health care utilization, and medications in a large managed care organization. Ann Allergy Asthma Immunol 2003;91:553-558.
25. Chen W, Mempel M, Schober W, Behrendt H, Ring J. Gender difference, sex hormones, and immediate type hypersensitivity reactions. Allergy 2008;63:1418-1427.
26. Carroll KN, Griffin MR, Gebretsadik T, Shintani A, Mitchel E, Hartert TV. Racial differences in asthma morbidity during pregnancy. Obstet Gynecol 2005;106:66-72.
27. Pesole G, Grillo G, Larizza A, Liuni S. The untranslated regions of eukaryotic mRNAs: structure, function, evolution and bioinformatic tools for their analysis. Brief Bioinform 2000;1:236-249.
28. Laprise C. The Saguenay-Lac-Saint-Jean asthma familial collection: the genetics of asthma in a young founder population. Genes Immun 2014;15:247.
29. Stenton GR, Ulanova M, Déry RE, Merani S, Kim MK, Gilchrist M, Puttagunta L, Musat-Marcu S, James D, Schreiber AD, Befus AD. Inhibition of allergic inflammation in the airways using aerosolized antisense to Syk kinase. J Immunol 2002;169:1028-1036.
30. Toyama T, Iwase H, Yamashita H, Hara Y, Omoto Y, Sugiura H, Sugiura H, Zhang Z, Fujii Y. Reduced expression of the Syk gene is correlated with poor prognosis in human breast cancer. Cancer Lett 2003;189:97-102.
31. Kepley CL, Youssef L, Andrews RP, Wilson BS, Oliver JM. Multiple defects in Fc epsilon RI signaling in Syk-deficient nonreleaser basophils and IL-3-induced recovery of Syk expression and secretion. J Immunol 2000;165:5913-5920.
32. Andrews RP, Kepley CL, Youssef L, Wilson BS, Oliver JM. Regulation of the very late antigen-4-mediated adhesive activity of normal and nonreleaser basophils: roles for Src, Syk, and phosphatidylinositol 3-kinase. J leukocyte Biol 2001;70:776-782.
33. Havard S, Scola AM, Kay LJ, Ishmael SS, MacGlashan DW, Jr., Peachell PT. Characterization of syk expression in human lung mast cells: relationship with function. Clin Exp Allergy 2011;41:378-388.
34. Wong WS. Inhibitors of the tyrosine kinase signaling cascade for asthma. Curr Opin Pharmacol 2005;5:264-271.
35. Barnes PJ. New therapies for asthma. Trends Mol Med 2006;12:515-520.
36. Youssef LA, Schuyler M, Wilson BS, Oliver JM. Roles for the high affinity IgE receptor, fcepsilon RI, of human basophils in the pathogenesis and therapy of allergic asthma: Disease Promotion, Protection or Both? Open Allergy J 2010;3:91-101.

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