Tissue transglutaminase-mediated AT1 receptor sensitization underlies pro-inflammatory cytokine LIGHT-induced hypertension.

BACKGROUND Although numerous recent studies have shown a strong link between inflammation and hypertension, the underlying mechanisms by which inflammatory cytokines induce hypertension remain to be fully elucidated. Hypertensive disorders are also associated with elevated pressor sensitivity. Tissue transglutaminase (TG2), a potent cross-linking enzyme, is known to be transcriptionally activated by inflammatory cytokines and stabilize angiotensin II (Ang II) receptor AT(1) (AT(1)R) via ubiquitination-preventing posttranslational modification. Here we sought to investigate the TG2-mediated AT(1)R stabilization in inflammation-induced hypertension and its functional consequences with a focus on receptor abundance and Ang II responsiveness. METHODS AND RESULTS Using an experimental model of inflammation-induced hypertension established by introducing the pro-inflammatory tumor necrosis factor cytokine LIGHT, we provide pharmacologic and genetic evidence that TG2 is required for LIGHT-induced hypertension (systolic pressure on day 6: LIGHT = 152.3 +/- 7.4 vs. LIGHT+ERW1041E [TG2 inhibitor] = 105.8 +/- 13.1 or LIGHT+TG2(-/-) = 114.3 +/- 4.3 mm Hg, P < 0.05, n = 4-5) and renal compromise (urine albumin/creatinine: LIGHT = 0.17 +/- 0.05 vs. LIGHT+ERW1041E = 0.03 +/- 0.01 or LIGHT+TG2(-/-) = 0.06 +/- 0.01 mg/mg; plasma creatinine: LIGHT = 1.11 +/- 0.04 vs. LIGHT+ERW1041E = 0.94 +/- 0.04 or LIGHT+TG2(-/-) = 0.88 +/- 0.09 mg/dl; urine volume: LIGHT = 0.23 +/- 0.1 vs. LIGHT+ERW1041E = 0.84 +/- 0.13 or LIGHT+TG2(-/-) = 1.02 +/- 0.09 ml/24 hour on day 14, P < 0.05, n = 4-5). Our mechanistic studies showed that the TG2-mediated AT(1)R modification and accumulation (relative renal AT(1)R level: phosphate-buffered saline [PBS] = 1.23 +/- 0.22, LIGHT = 3.49 +/- 0.37, and LIGHT+ERW1041E = 1.77 +/- 0.46, P < 0.05, n = 3; LIGHT+TG2(+/+)= 85.28 +/- 36.11 vs. LIGHT+TG2(-/-) = 7.01 +/- 5.68, P < 0.05, n = 3) induced by LIGHT is associated with abrogated beta-arrestin binding (AT(1)R/associated beta-arrestin ratio: PBS = 2.62 +/- 1.07, LIGHT = 38.60 +/- 13.91, and LIGHT+ERW1041E = 6.97 +/- 2.91, P < 0.05, n = 3; LIGHT+TG2(+/+)= 66.43 +/- 44.81 vs. LIGHT+TG2(-/-) = 2.45 +/- 1.78, P < 0.01, n = 3) and could be found in renal medulla tubules of kidneys (relative tubular AT1R level: PBS = 5.91 +/- 2.93, LIGHT = 92.82 +/- 19.54, LIGHT+ERW1041E = 28.49 +/- 11.65, and LIGHT+TG2(-/-) = 0.14 +/- 0.10, P < 0.01, n = 5) and the blood vasculature (relative vascular AT1R level: PBS = 0.70 +/- 0.30, LIGHT = 13.75 +/- 2.49, and LIGHT+ERW1041E = 3.28 +/- 0.87, P < 0.01, n = 3), 2 of the tissues highly related to the genesis of hypertension. Our in vitro cellular assays showed that LIGHT stimulation triggered a rapid TG2-dependent increase in the abundance of AT(1)Rs (relative AT(1)R level after 2-hour LIGHT treatment: AT(1)R (WT)+TG2 = 2.21 +/- 0.23, AT(1)R (Q315A)+TG2 = 0.18 +/- 0.23, P < 0.05 vs. starting point = 1, n = 2) and downstream calcium signaling (fold increase in NFAT-driven luciferase activity: Saline = 0.02 +/- 0.03, Ang II = 0.17 +/- 0.08, LIGHT = 0.05 +/- 0.04, LIGHT+Ang II = 0.90 +/- 0.04 (P < 0.01 vs. Ang II), and LIGHT+Ang II+ERW1041E = 0.15 +/- 0.15 (P < 0.01 vs. LIGHT+Ang II), n = 3). CONCLUSIONS Our data indicate an essential and systemic role for TG2 in bridging inflammation to hypertension via its posttranslational modifications stabilizing AT(1) receptor and sensitizing Ang II. Our findings also suggest that TG2 inhibitors could be used as a novel group of cardiovascular agents.