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Katharina Rump, Theresa Spellenberg, Alexander von Busch, Alexander Wolf, Dominik Ziehe, Patrick Thon, Tim Rahmel, Michael Adamzik, Björn Koos, Matthias Unterberg

• The quantity of aquaporin 5 protein in neutrophil granulocytes is associated with human sepsis-survival. The C-allele of the aquaporin (\(\it AQP5\))-1364A/C polymorphism was shown to be associated with decreased AQP5 expression, which was shown to be relevant in this context leading towards improved outcomes in sepsis. To date, the underlying mechanism of the C-allele—leading to lower AQP5 expression—has been unknown. Knowing the detailed mechanism depicts a crucial step with a target to further interventions. Genotype-dependent regulation of AQP5 expression might be mediated by the epigenetic mechanism of promoter methylation and treatment with epigenetic-drugs could maybe provide benefit. Hence, we tested the hypothesis that \(\it AQP5\) promoter methylation differs between genotypes in specific types of immune cells.: \(\it AQP5\) promoter methylation was quantified in cells of septic patients and controls by methylation-specific polymerase chain reaction and quantified by a standard curve. In cell-line models, AQP5 expression was analyzed after demethylation to determine the impact of promoter methylation on AQP5 expression. C-allele of \(\it AQP5\)-1364 A/C promoter polymorphism is associated with a five-fold increased promoter methylation in neutrophils (\(\it p\) = 0.0055) and a four-fold increase in monocytes (\(\it p\) = 0.0005) and lymphocytes (\(\it p\) = 0.0184) in septic patients and healthy controls as well. In addition, a decreased \(\it AQP5\) promoter methylation was accompanied by an increased AQP5 expression in HL-60 (\(\it p\) = 0.0102) and REH cells (\(\it p\) = 0.0102). The C-allele which is associated with lower gene expression in sepsis is accompanied by a higher methylation level of the \(\it AQP5\) promoter. Hence, \(\it AQP5\) promoter methylation could depict a key mechanism in genotype-dependent expression.