Highlights
- •Cirrhosis is associated with TLR4-related sensitization to endotoxins and a switch from apoptotic to necroptotic cell death.
- •Inhibition of TLR4 signaling by TAK-242 reduces immune cell responses and hepatocyte injury in response to LPS in vitro.
- •In models of acute and acute-on-chronic liver failure, TAK-242 mitigated LPS-driven systemic inflammation and organ injury.
Background & Aims
Toll-like receptor 4 (TLR4) plays an essential role in mediating organ injury in acute
liver failure (ALF) and acute-on-chronic liver failure (ACLF). Herein, we assess whether
inhibiting TLR4 signaling can ameliorate liver failure and serve as a potential treatment.
Methods
Circulating TLR4 ligands and hepatic TLR4 expression were measured in plasma samples
and liver biopsies from patients with cirrhosis. TAK-242 (TLR4 inhibitor) was tested
in vivo (10 mg/kg intraperitoneally) in rodent models of ACLF (bile duct ligation + lipopolysaccharide
[LPS]; carbon tetrachloride + LPS) and ALF (galactosamine + LPS) and in vitro on immortalized human monocytes (THP-1) and hepatocytes (HHL5). The in vivo therapeutic effect was assessed by coma-free survival, organ injury and cytokine
release and in vitro by measuring IL-6, IL-1β or cell injury (TUNEL), respectively.
Results
In patients with cirrhosis, hepatic TLR4 expression was upregulated and circulating
TLR4 ligands were increased (p <0.001). ACLF in rodents was associated with a switch from apoptotic cell death in
ALF to non-apoptotic forms of cell death. TAK-242 reduced LPS-induced cytokine secretion
and cell death (p = 0.002) in hepatocytes and monocytes in vitro. In rodent models of ACLF, TAK-242 administration improved coma-free survival, reduced
the degree of hepatocyte cell death in the liver (p <0.001) and kidneys (p = 0.048) and reduced circulating cytokine levels (IL-1β, p <0.001). In a rodent model of ALF, TAK-242 prevented organ injury (p <0.001) and systemic inflammation (IL-1β, p <0.001).
Conclusion
This study shows that TLR4 signaling is a key factor in the development of both ACLF
and ALF; its inhibition reduces the severity of organ injury and improves outcome.
TAK-242 may be of therapeutic relevance in patients with liver failure.
Lay summary
Toll-like receptor 4 (or TLR4) mediates endotoxin-induced tissue injury in liver failure
and cirrhosis. This receptor sensitizes cells to endotoxins, which are produced by
gram-negative bacteria. Thus, inhibiting TLR4 signaling with an inhibitor (TAK-242)
ameliorates organ injury and systemic inflammation in rodent models of acute and acute-on-chronic
liver failure.
Graphical abstract
Graphical Abstract
Keywords
To read this article in full you will need to make a payment
Purchase one-time access:
Academic & Personal: 24 hour online accessCorporate R&D Professionals: 24 hour online accessOne-time access price info
- For academic or personal research use, select 'Academic and Personal'
- For corporate R&D use, select 'Corporate R&D Professionals'
Subscribe:
Subscribe to Journal of HepatologyAlready a print subscriber? Claim online access
Already an online subscriber? Sign in
Register: Create an account
Institutional Access: Sign in to ScienceDirect
References
- Acute-on-chronic liver failure is a distinct syndrome that develops in patients with acute decompensation of cirrhosis.Gastroenterology. 2013; 144 (1437 e1421-1429): 1426-1437
- Validation of CLIF-C ACLF score to define a threshold for futility of intensive care support for patients with acute-on-chronic liver failure.Crit Care. 2018; 22: 254
- Cirrhosis-associated immune dysfunction: distinctive features and clinical relevance.J Hepatol. 2014; 61: 1385-1396
- Systemic inflammatory response and serum lipopolysaccharide levels predict multiple organ failure and death in alcoholic hepatitis.Hepatology. 2015; 62: 762-772
- Recombinant alkaline phosphatase prevents acute on chronic liver failure.Sci Rep. 2020; 10: 389
- A model of acute kidney injury in mice with cirrhosis and infection.Liver Int. 2016; 36: 865-873
- Increased sensitivity to endotoxemia in the bile duct-ligated cirrhotic Rat.Hepatology. 1999; 30: 1198-1205
- Acute liver failure.N Engl J Med. 2013; 369: 2525-2534
- Acute liver failure: a review.Clin Liver Dis. 2006; 10 (vii-viii): 239-258
- Ethyl pyruvate prevents inflammatory factors release and decreases intestinal permeability in rats with D-galactosamine-induced acute liver failure.Hepatobiliary Pancreat Dis Int. 2013; 12: 180-188
- Curcumin alleviates lipopolysaccharide induced sepsis and liver failure by suppression of oxidative stress-related inflammation via PI3K/AKT and NF-kappaB related signaling.Biomed Pharmacother. 2016; 83: 302-313
- Thymosin alpha1 treatment reduces hepatic inflammation and inhibits hepatocyte apoptosis in rats with acute liver failure.Exp Ther Med. 2018; 15: 3231-3238
- Defective LPS signaling in C3H/HeJ and C57BL/10ScCr mice: mutations in Tlr4 gene.Science. 1998; 282: 2085-2088
- TLR4-HMGB1-, MyD88- and TRIF-dependent signaling in mouse intestinal ischemia/reperfusion injury.World J Gastroenterol. 2015; 21: 8314-8325
- Changes in expression of the membrane receptors CD14, MHC-II, SR-A, and TLR4 in tissue-specific monocytes/macrophages following porphyromonas gingivalis-LPS stimulation.Inflammation. 2018; 41: 418-431
- Beta2-integrin-induced p38 MAPK activation is a key mediator in the CD14/TLR4/MD2-dependent uptake of lipopolysaccharide by hepatocytes.J Biol Chem. 2008; 283: 29433-29446
- TAK-242 (resatorvid), a small-molecule inhibitor of Toll-like receptor (TLR) 4 signaling, binds selectively to TLR4 and interferes with interactions between TLR4 and its adaptor molecules.Mol Pharmacol. 2011; 79: 34-41
- Prevention of acute kidney injury in a rodent model of cirrhosis following selective gut decontamination is associated with reduced renal TLR4 expression.J Hepatol. 2012; 56: 1047-1053
- Role of toll-like receptor 4 in mediating multiorgan dysfunction in mice with acetaminophen induced acute liver failure.Liver Transpl. 2013; 19: 751-761
- Pharmacological inhibition of toll-like receptor-4 signaling by TAK242 prevents and induces regression of experimental organ fibrosis.Front Immunol. 2018; 9: 2434
- Positive feedback regulation between transglutaminase 2 and toll-like receptor 4 signaling in hepatic stellate cells correlates with liver fibrosis post schistosoma japonicum infection.Front Immunol. 2017; 8: 1808
- Inhibition of Toll-like receptor 4 ameliorates experimental postischemic injury in the cholestatic liver through inhibition of high-mobility group box protein b1 (HMGB1) signaling.Surgery. 2018; 163: 270-276
- Inhibition of Toll-like receptor 4 suppresses liver injury induced by biliary obstruction and subsequent intraportal lipopolysaccharide injection.Am J Physiol Gastrointest Liver Physiol. 2014; 306: G244-G252
- TAK-242 treatment ameliorates liver ischemia/reperfusion injury by inhibiting TLR4 signaling pathway in a swine model of Maastricht-category-III cardiac death.Biomed Pharmacother. 2016; 84: 495-501
- Bacterial infections in cirrhosis: a position statement based on the EASL Special Conference 2013.J Hepatol. 2014; 60: 1310-1324
- Acute-on-chronic liver failure: a new syndrome that will re-classify cirrhosis.J Hepatol. 2015; 62: S131-S143
- Pivotal advance: vasoactive intestinal peptide inhibits up-regulation of human monocyte TLR2 and TLR4 by LPS and differentiation of monocytes to macrophages.J Leukoc Biol. 2007; 81: 893-903
- Nuclear factor kappaB up-regulation of CCAAT/enhancer-binding protein beta mediates hepatocyte resistance to tumor necrosis factor alpha toxicity.Hepatology. 2010; 52: 2118-2126
- Hepatoprotective and anti-inflammatory cytokines in alcoholic liver disease.J Gastroenterol Hepatol. 2012; 27: 89-93
- Role of Toll-like receptors 2, 4, and 9 in mediating neutrophil dysfunction in alcoholic hepatitis.Am J Physiol Gastrointest Liver Physiol. 2009; 296: G15-G22
- Toll-like receptor-4 signaling and Kupffer cells play pivotal roles in the pathogenesis of non-alcoholic steatohepatitis.J Hepatol. 2007; 47: 571-579
- Deficiency in myeloid differentiation factor-2 and toll-like receptor 4 expression attenuates nonalcoholic steatohepatitis and fibrosis in mice.Am J Physiol Gastrointest Liver Physiol. 2011; 300: G433-G441
- TLR4 enhances TGF-beta signaling and hepatic fibrosis.Nat Med. 2007; 13: 1324-1332
- Role of toll-like receptors in immune activation and tolerance in the liver.Front Immunol. 2014; 5: 221
- Toll-like receptor 4 mediates inflammatory signaling by bacterial lipopolysaccharide in human hepatic stellate cells.Hepatology. 2003; 37: 1043-1055
- A randomized, double-blind, placebo-controlled trial of TAK-242 for the treatment of severe sepsis.Crit Care Med. 2010; 38: 1685-1694
- Both intrinsic and extrinsic apoptotic pathways are involved in Toll-like receptor 4 (TLR4)-induced cell death in monocytic THP-1 cells.Immunobiology. 2017; 222: 198-205
- Inhibition of apoptosis protects mice from ethanol-mediated acceleration of early markers of CCl4 -induced fibrosis but not steatosis or inflammation.Alcohol Clin Exp Res. 2012; 36: 1139-1147
- A positive feedback loop between RIP3 and JNK controls non-alcoholic steatohepatitis.EMBO Mol Med. 2014; 6: 1062-1074
- Cell death and cell death responses in liver disease: mechanisms and clinical relevance.Gastroenterology. 2014; 147: 765-783 e764
- Live to die another way: modes of programmed cell death and the signals emanating from dying cells.Nat Rev Mol Cell Biol. 2015; 16: 329-344
- Role of necroptosis in the pathogenesis of solid organ injury.Cell Death Dis. 2015; 6: e1975
- The chemical inhibitors of cellular death, PJ34 and Necrostatin-1, down-regulate IL–33 expression in liver.J Mol Med (Berl). 2015; 93: 867-878
- Dysfunction of circulating polymorphonuclear leukocytes and monocytes in ambulatory cirrhotics predicts patient outcome.Dig Dis Sci. 2016; 61: 2294-2302
- Functional contribution of elevated circulating and hepatic non-classical CD14CD16 monocytes to inflammation and human liver fibrosis.PLoS One. 2010; 5: e11049
Article info
Publication history
Published online: January 24, 2020
Accepted:
January 14,
2020
Received in revised form:
December 22,
2019
Received:
October 9,
2019
Footnotes
Author names in bold designate shared co-first authorship
Identification
Copyright
© 2020 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.
