Recombinant Thrombomodulin Modulates Murine Colitis Possibly via High-Mobility Group Box 1 Protein Inhibition

Abstract

Background and Aim: Thrombomodulin (TM) is an anticoagulant cofactor protein. We hypothesized that its recombinant soluble TM (rhTM) form, widely used to treat disseminated intravascular coagulation, might have anti-inflammatory action in inflammatory bowel disease (IBD), possibly through its inhibition of high-mobility group box 1 protein (HMGB1). Methods: We investigated inflammatory effects of HMGB1 and anti-inflammatory effect of rhTM in dextran sulfate sodium (DSS)-treated mice, some cell lines and ulcerative colitis (UC) patients, particularly focusing on changes of vascular endothelial adhesion molecules. Results: Treatments with rhTM significantly attenuated DSS-treated mice clinically and histologically. The mRNA levels of proinflammatory cytokines and adhesion molecules were decreased by rhTM. Increased inflammatory cells in the colonic mucosa strongly expressed HMGB1 in the cytoplasm in the DSS-treated mice and UC patients' colonic mucosa, which were significantly decreased by rhTM in mice. In in vitro experiments, rhTM significantly decreased the mRNA levels of tumor necrosis factor-alpha (TNF-α) and adhesion molecules increased by endotoxin exposures in RAW 264.7 (macrophage cell line) and bEND.3 cells (endothelial cell line), suggesting the proinflammatory role of HMGB1 in TNF-α production from macrophages. Conclusions: These findings suggest that rhTM may be useful for the treatment of IBD by attenuating inflammatory cytokine production and adhesion molecule expression, partly because of its inhibition of HMGB1.

Recombinant human soluble thrombomodulin and mortality in sepsis-induced disseminated intravascular coagulation

“…Furthermore, unlike recombinant human-activated protein C, the anti-coagulant effects of rhTM are dependent on thrombin activity in the systemic circulation, as rhTM cannot activate protein C after the attenuation of thrombin generation (2,29). In addition to its anticoagulant activities, the anti-inflammatory effects of rhTM include the inhibition of leukocyte adhesion to endothelial cells, inhibition of complement pathways, neutralization of lipopolysaccharides, suppression of inflammatory cytokines, and degradation of high-mobility group box 1 protein (33,34). Thus, rhTM administration could be considered appropriate treatment for patients with sepsis-induced DIC.…”

Role of high‐mobility group box 1 and its modulation by thrombomodulin/thrombin axis in neuropathic and inflammatory pain

“…Serum HMGB1 levels increases in mice with colitis induced by dextran sulfate sodium (DSS), and systemic administration of the anti‐HMGB1‐neutralizing antibody prevent the DSS‐induced colitis (Yamasaki et al, 2009). Inactivation of HMGB1 with systemic administration of TMα or glycyrrhizate (an anionic salt of glycyrrhizin) or intracolonic administration of adeno‐associated virus‐mediated colonic secretory expression of HMGB1 box A reduces the DSS‐induced colitis (Ueda et al, 2015; Vitali et al, 2013; Zheng et al, 2016). Most interestingly, fecal HMGB1 levels in patients with Crohn's disease or ulcerative colitis, but not irritable bowel syndrome (IBS), are much higher than the healthy control, and serum and fecal HMGB1 levels increase in TNBS‐induced colitis mice (Chen et al, 2019).…”

“…We have shown that intraplantar injection of TMα suppresses the long-lasting allodynia following intraplantar injection of at-HMGB1, ds-HMGB1 or LPS in mice, an effect reversed by i.p. argatroban, a thrombin inhibitor, and promoted by intraplantar thrombin (Tsujita et al, 2018). The recombinant human TM D1-D2-D3 can also be generated by yeast and this protein is possibly different from the TMα generated by dose of oxaliplatin (Tsubota et al, 2019), suggesting that the endothelial TM/thrombin axis potentially functions to reduce the increased HMGB1 in the blood stream and the development of CIPN following treatment with chemotherapeutics including oxaliplatin (Figure 3).…”

“…Injection of HMGB1 to the perisciatic nerve or hindpaw causes mechanical hyperalgesia and allodynia, as assessed by a paw pressure test and von Frey test, respectively, and thermal hyperalgesia, as assessed by a hot plate test or radiant heat-induced plantar test, most probably by activating RAGE and/or TLR4 in rats (Nishida et al, 2016;Shibasaki et al, 2010;Tanaka et al, 2013; Table 1). In mice, intraplantar administration of at-HMGB1 and ds-HMGB1 produces long-lasting mechanical allodynia via RAGE and TLR4, respectively, and the latter form of HMGB1 is 10-fold more potent than the former (Hayashi et al, 2018;Tsujita et al, 2018;Yamasoba et al, 2016). There is evidence that TLR5 is involved in the intraplantar HMGB1-induced mechanical allodynia in mice (Das et al, 2016).…”

“…HMGB1 is now considered to play a pronociceptive role in the CNS and peripheral tissue and to serve as a therapeutic target for the treatment of pathological pain (Agalave & Svensson, 2015;Tanaka et al, 2013;Yamasoba et al, 2016). Accumulating evidence from preclinical studies indicates that exogenously applied TMα reduces HMGB1-dependent pathological pain including chemotherapy-induced peripheral neuropathy (CIPN) and that the endogenous TM/thrombin system potentially functions to relieve CIPN (Hayashi et al, 2018;Irie et al, 2017;Nishida et al, 2016;Sekiguchi et al, 2018;Tanaka et al, 2013;Tanaka et al, 2014;Tsubota et al, 2019;Tsujita et al, 2018). A placebo-controlled, randomized, double-blind phase II study has shown the promising efficacy of TMα in delaying and reducing CIPN in colon cancer patients undergoing adjuvant chemotherapy with an oxaliplatin-based regimen (T. Kato et al, 2018).…”