Genetic Variants of Intron Region of Aquaporin AQP5 Gene and Development of Pulmonary Edema in Lung Infection Complicated by Septic Shock

Purpose of the study. Determine the value of genetic variants of a single nucleotide polymorphic site rs3736309 of intron 3 of aquaporin5 (AQP5) gene in the course of critical illness in patients with documented pulmonary infection. Materials and methods. Patients with critical illness admitted to the intensive care units were examined during the course of treatment (n=86, age 27 to 82 years, mean age 53.20±14.34 years). Main diagnosis included malignancies (15%), peritonitis (16%) and necrotizing pancreatitis (37%). Patients developed nosocomial pneumonia (55%), acute respiratory distress syndrome (ARDS) (54%), septic shock (48%), ARDS combined with septic shock (33%). Bacterial species of Pseudomonas aeruginosa , Klebsiella pneumoniae, Acinetobacter baumannii, and/or Proteus mirabilis alone or in association were revealed in lavage fluid. DNA genotyping DNA was carried out using tetraprimer polymerase chain reaction (PCR). Statistical processing was performed using GraphPad InStat program (GraphPad, USA).

Results. The distribution of frequencies of genotypes AA, GA and GG (AQP5, rs3736309) in cohort of patients corresponded to HardyWeinberg equilibrium (P=0.923) and was similar to frequencies of same alleles determined in a conditionally healthy Caucasian individuals (literature data) (P>0.05). In a subgroup of patients with septic shock and AQP5 AA (rs3736309) genotype the lower EVLWI values were found compared to patients with genotypes GG and GA with septic shock in spite of the same approach to treatment. The differences between genetically different subgroups of patients with septic shock were maintained throughout the life of the survey (P<0.05,days 1, 3, 5 and 7). Genetic variant AQP5 G+ (rs3736309) contributed to the development of pulmonary edema resistant to treatment (odds ratio, OR = 6,75; P=0.032). Only the subgroup of patients with septic shock and genotype G + (but not all patients or the subgroup of patients without septic shock of the same genotype) were characterized by significantly elevated levels of surfactant protein SPD in plasma compared to patients of genotype AQP5 AA with septic shock (P<0.05).

Conclusion. In septic shock, the presence of homozygous variant allele A (AA) of AQP5 rs3736309 is a favorable factor for patients developing the pulmonary edema. The presence of allele AQP5 G (rs3736309) is a risk factor for developing severe pulmonary edema and unfavorable prognosis in spite of treatment.

1. Smelaya T.V., Kuzovlev A.N., Moroz V.V., Golubev A.M., Belopolskaya O.B., Salnikova L.E. Molekulyarnogeneticheskie markery nozokomialnoi pnevmonii ostrogo respiratornogo distresssindroma. Obshchaya Reanimatologiya. [Search for common molecular genetic markers of nosocomial pneumonia and acute respiratory distress syndrome. General Reanimatology]. 2015; 11 (3): 24—38. http://dx.doi.org/10.15360/18139779201532438. [In Russ.]

2. Cardoso C.P., Oliveira A.J., Botoni F.A., Rezende I.C., AlvesFilho J.C., Cunha F.Q., Estanislau J.A., Magno L.A., RiosSantos F. Interleukin10 rs2227307 and CXCR2 rs1126579 polymorphismsmodulate the predis position to septic shock. Mem. Inst. Oswaldo Cruz. 2015; 110 (4): 453—460. http://dx.doi.org/10.1590/007402760150003. PMID: 26038959

3. Sotos-Prieto M., Peñalvo J.L. Genetic variation of apolipoproteins, diet and other environmental interactions; an updated review. Nutr. Hosp. 2013; 28 (4): 999—1009. http://dx.doi.org/10.3305/nh.2013. 28.4.6475. PMID: 23889614

4. PerezMartinez P., GarciaRios A., DelgadoLista J., PerezJimenez F., LopezMiranda J. Nutrigenetics of the postprandial lipoprotein metabolism: evidences from human intervention studies. Curr. Vasc. Pharmacol. 2011; 9 (3): 287—291. http://dx.doi.org/10.2174/157016111795495495. PMID: 21314629

5. Day R.E., Kitchen P., Owen D.S., Bland C., Marshall L., Conner A.C., Bill R.M., Conner M.T. Human aquaporins: regulators of transcellular water flow. Biochim. Biophys. Acta. 2014; 1840 (5): 1492—1506. http://dx.doi.org/10.1016/j.bbagen.2013.09.033. PMID: 24090884

6. Verkman A.S. More than just water channels: unexpected cellular roles of aquaporins. J. Cell Sci. 2005; 118 (Pt 15): 3225—3232. http://dx.doi.org/10.1242/jcs.02519. PMID: 16079275

7. Mezzasoma L., Cagini L., Antognelli C., Puma F., Pacifico E., Talesa V.N. TNFα regulates natriuretic peptides and aquaporins in human bronchial epithelial cells BEAS2B. Mediators Inflamm. 2013; 2013: 159349. http://dx.doi.org/10.1155/2013/159349. PMID: 24369440

8. Bloch O., Manley G.T. The role of aquaporin4 in cerebral water transport and edema. Neurosurg. Focus. 2007; 22 (5): E3. http://dx.doi.org/10.3171/foc.2007.22.5.4. PMID: 17613234

9. Li J., Xu M., Fan Q., Xie X., Zhang Y., Mu D., Zhao P., Zhang B., Cao F., Wang Y., Jin F., Li Z. Tanshinone IIA ameliorates seawater exposure induced lung injury by inhibiting aquaporins (AQP) 1 and AQP5 expression in lung. Respir. Physiol. Neurobiol. 2011; 176 (1—2): 39—49. http://dx.doi.org/10.1016/j.resp.2011.01.005. PMID: 21244858

10. Ning Y., Ying B., Han S., Wang B., Wang X., Wen F. Polymorphisms of aquaporin5 gene in chronic obstructive pulmonary disease in a Chinese population. Swiss Med. Wkly. 2008; 138 (39—40): 573—578. http://dx.doi.org/10.2008/39/smw12240. PMID: 18853286

11. Adamzik M., Frey U.H., Möhlenkamp S., Scherag A., Waydhas C., Marggraf G., Dammann M., Steinmann J., Siffert W., Peters J. Aquaporin 5 gene promoter 1364A/C polymorphism associated with 30day survival in severe sepsis. Anesthesiology. 2011; 114 (4): 912—917. http://dx.doi.org/10.1097/ALN.0b013e31820ca911. PMID: 21427539

12. Dellinger R.P., Levy M.M., Rhodes A., Annane D., Gerlach H., Opal S.M., Sevransky J.E., Sprung C.L., Douglas I.S., Jaeschke R., Osborn T.M., Nunnally M.E., Townsend S.R., Reinhart K., Kleinpell R.M., Angus D.C., Deutschman C.S., Machado F.R., Rubenfeld G.D., Webb S.A., Beale R.J., Vincent J.L., Moreno R.; Surviving Sepsis Campaign Guidelines Committee including the Pediatric Subgroup. Surviving sepsis campaign: international guidelines for management of severe sepsis and septic shock: 2012. Crit. Care Med. 2013; 41 (2): 580—637. http://dx.doi.org/ 10.1007/s0013401227698. PMID: 23353941

13. Moroz V.V., Golubev A.M., Kuzovlev A.N. Otek legkikh: klassifikatsiya, mekhanizmy razvitiya, diagnostika. Obshchaya Reanimatologiya. [Pulmonary edema: classification, mechanisms of development, diagnosis. General Reanimatology]. 2009; 5 (1): 83—88. http://dx.doi.org/10.15360/181397792009183. [In Russ.]

14. Tablan O.C., Anderson L.J., Besser R., Bridges C., Hajjeh R.; CDC; Healthcare Infection Control Practices Advisory Committee. Guidelines for preventing healthcare—associated pneumonia, 2003: recommenda tions of CDC and the Healthcare Infection Control Practices Advisory Committee. MMWR Recomm Rep. 2004; 53 (RR3): 1—36. PMID:15048056

15. Gelfand B.R., Yakovlev S.V., Protsenko D.N., Belotserkovsky B.Z. Nozokomialnaya pnevmoniya, svyazannaya s iskusstvennoi ventilyat siei legkikh: vozmozhna li standartizatsiya terapii? [Ventilatorassociated nosocomial pneumonia: is there a possibility to standartoze treatment?] Consilium Medicum. 2004; 6 (4): 245—248. [In Russ.]

16. Pugin J., Auckenthaler R., Mili N., Janssens J.P., Lew P.D., Suter P.M. Diagnosis of ventilator associated pneumonia by bacteriological analysis of bronchoscopic and non bronchoscopic «blind» bronchoalveolar lavage fiuid. Am. Rev. Respir. Dis. 1991; 143 (5Pt 1): 1121—1129. http://dx.doi.org/10.1164/ajrccm/143.5_Pt_1.1121. PMID: 2024824

17. Chuchalin A.G., Gelfand B.R. (red.). Nozokomialnaya pnevmoniya u vzroslykh. Rossiiskie natsionalnye rekomendatsii. [Nosocomial pneumonia in adults. Russian national guidelines]. Moscow; 2009: 92. [In Russ.]

18. Moroz V.V., Golubev A.M., Kuzovlev A.N., Pisarev V.M. Novye diagnosticheskie kandidatnye molekulyarnye biomarkery ostrogo respiratornogo distresssindroma. Obshchaya Reanimatologiya. [New diagnostic candidate molecular biomarkers of acute respiratory distress syndrome. General Reanimatology]. 2014; 10 (4): 6—10. http://dx.doi.org/10.15360/1813977920144610. [In Russ.]

19. http://www.ncbi.nlm.nih.gov/projects/SNP

20. Newton C.R., Heptinstall L.E., Summers C., Super M., Schwarz M., Anwar R., Graham A., Smith J.C., Markham A.F. Amplification refractory mutation system for prenatal diagnosis and carrier assessment in cystic fibrosis. Lancet. 1989; 2 (8678—8679): 1481—1483. http://dx.doi.org/10.1016/S01406736(89)929310. PMID: 2574768

21. Hamajima N., Katsuda N., Matsuo K., Saito T., Ito L.S., Ando M., Inoue M., Takezaki T., Tajima K. Smoking habit and interleukin lB C31T polymorphism. J. Epidemiol. 2001; 11 (3): 120—125. PMID: 11434423

22. Salnikova L.E., Chumachenko A.G., Vesnina I.N., Lapteva N.Sh., Kuznetsova G.I., Abilev S.K., Rubanovich A.V. Polimorfizm genov reparatsii i tsitogeneticheskie effekty oblucheniya. [Polymorphism of repair genes and cytogenetic radiation effects]. Radiatsionnaya Biologiya. Radioekologiya. 2010; 50 (6): 656—662. PMID: 21434392. [In Russ.]

23. Egorova I.N., Vlasenko A.V., Moroz V.V., Yakovlev V.N., Alekseyev V.G. Ventilyatorassotsiirovannaya pnevmoniya: diagnostika, profilaktika, lechenie (sovremennoe sostoyanie voprosa). Obshchaya Reanimatologiya. [Ventilatorassociated pneumonia: diagnosis, prevention, treatment (stateoftheart of the problem). General Reanimatology]. 2010; 6 (1): 79—88. http://dx.doi.org/10.15360/181397792010179. [In Russ.]

24. Pretto E.A. (ed.). Oxford textbook of transplant anaesthesia and critical care. USA: Oxford University Press; 450.

25. Agre P., King L.S., Yasui M., Guggino W.B., Ottersen O.P., Fujiyoshi Y., Engel A., Nielsen S. Aquaporin water channels—from atomic structure to clinical medicine. J. Physiol. 2002; 542 (Pt 1): 3—16. http://dx.doi.org/10.1113/jphysiol.2002.020818. PMID: 12096044

26. Agre P., Kozono D. Aquaporin water channels: molecular mechanisms for human diseases. FEBS Lett. 2003; 555 (1): 72—78. http://dx.doi.org/10.1016/S00145793(03)010834. PMID: 14630322

27. Shen Y., Wang X., Wang Y., Wang X., Chen Z., Jin M., Bai C. Lipopolysaccharide decreases aquaporin 5, but not aquaporin 3 or aquaporin 4, expression in human primary bronchial epithelial cells. Respirology. 2012; 17 (7): 1144—1149. http://dx.doi.org/10.1111/j.14401843.2012.02228.x. PMID: 22809117

28. Yang B., Verkman A.S. Analysis of double knockout mice lacking aqua porin1 and urea transporter UTB. Evidence for UTBfacilitated water transport in erythrocytes. J. Biol. Chem. 2002; 277 (39): 36782—36786. http://dx.doi.org/10.1074/jbc.M206948200. PMID: 12133842

29. Sun Y., Sun L., Liu S., Song J., Cheng J., Liu J. Effect of emodin on Aquaporin 5 expression in rats with sepsisinduced acute lung injury. J. Tradit. Chin. Med. 2015; 35 (6): 679—684. PMID: 26742314

30. Jin Y.,Yu G., Peng P., Zhang Y., Xin X. Downregulated expression of AQP5 on lung in rat DIC model induced by LPS and itseffect on the develop ment of pulmonary edema. Pulm. Pharmacol. Ther. 2013; 26 (6): 661—665. http://dx.doi.org/10.1016/j.pupt.2013.03.013. PMID: 23538169

31. Jiang Y.X., Dai Z.L., Zhang X.P., Zhao W., Huang Q., Gao L.K. Dexmedetomidine alleviates pulmonary edema by upregulating AQP1 and AQP5expression in rats with acute lung injury induced by lipopolysaccharide. J. Huazhong Univ. Sci. Technolog. Med. Sci. 2015; 35 (5): 684—688. http://dx.doi.org/10.1007/s1159601514906. PMID:26489622

32. Moon C., Rousseau R., Soria J.C., Hoque M.O., Lee J., Jang S.J., Trink B., Sidransky D., Mao L. Aquaporin expression in human lymphocytes and dendritic cells. Am. J. Hematol. 2004; 75 (3): 128—133. http://dx.doi.org/10.1002/ajh.10476. PMID: 14978691

33. Shen Y., Wang Y., Chen Z., Wang D., Wang X., Jin M., Bai C. Role of aqua porin 5 in antigeninduced airway inflammation and mucous hyperpro duction in mice. J. Cell Mol. Med. 2011; 15 (6): 1355—1363. http://dx.doi.org/10.1111/j.15824934.2010.01103.x. PMID: 20550619

34. Wang G.F., Dong C.L., Tang G.S., Shen Q., Bai C.X. Membrane water permeability related to antigenpresenting function of dendritic cells. Clin. Exp. Immunol. 2008; 153 (3): 410—419. http://dx.doi.org/10.1111/j.13652249.2008.03702.x. PMID: 18647319

35. Mueller J.C. Linkage disequilibrium for different scales and applications. Brief Bioinform. 2004; 5 (4): 355—364. http://dx.doi.org/10.1093/bib/5.4.355. PMID: 15606972

36. Hansel N.N., Sidhaye V., Rafaels N.M., Gao L., Gao P., Williams R., Connett J.E., Beaty T.H., Mathias R.A., Wise R.A., King L.S., Barnes K.C. Aquaporin 5 polymorphisms and rate of lung function decline in chronic obstructive pulmonary disease. PLoS One. 2010; 5 (12): e14226. http://dx.doi.org/10.1371/journal.pone.0014226. PMID: 21151978

37. Llopart A., Comeron J.M., Brunet F.G., Lachaise D., Long M. Intron presenceabsence polymorphism in Drosophila driven by positive Darwinian selection. Proc. Natl. Acad. Sci. U S A. 2002; 99 (12): 8121—8126. http://dx.doi.org/10.1073/pnas.122570299. PMID: 12060758