Pan-caspase inhibitor F573 mitigates liver ischemia reperfusion injury in a murine model

Autoři: Mariusz Bral aff001;  Rena Pawlick aff001;  Braulio Marfil-Garza aff001;  Nidheesh Dadheech aff001;  Joshua Hefler aff001;  Aducio Thiesen aff002;  A. M. James Shapiro aff001
Působiště autorů: Department of Surgery, University of Alberta, Edmonton, Canada aff001;  Department of Pathology, University of Alberta, Edmonton, Canada aff002
Vyšlo v časopise: PLoS ONE 14(11)
Kategorie: Research Article
doi: 10.1371/journal.pone.0224567



Liver ischemia reperfusion injury (IRI) remains a challenge in liver transplantation. A number of compounds have previously demonstrated efficacy in mitigating IRI. Herein, we applied three specific additive strategies to a mouse IRI screening model to determine their relative potencies in reducing such injury, with a view to future testing in a large animal and clinical ex situ normothermic perfusion setting: 1) F573, a pan-caspase inhibitor, 2) anti-inflammatory anakinra and etanrecept and 3) BMX-001, a mimetic of superoxide dismutase.


A non-lethal liver ischemia model in mice was used. Additives in the treatment groups were given at fixed time points before induction of injury, compared to a vehicle group that received no therapeutic treatment. Mice were recovered for 6 hours following the ischemic insult, at which point blood and tissue samples were obtained. Plasma was processed for transaminase levels. Whole liver tissue samples were processed for histology, markers of apoptosis, oxidative stress, and cytokine levels.


In an in vivo murine IRI model, the F573 treatment group demonstrated statistically lower alanine aminotransferase (ALT) levels (p = 0.01), less evidence of apoptosis (p = 0.03), and lower cytokine levels compared to vehicle. The etanercept with anakinra treatment group demonstrated significantly lower cytokine levels. The BMX-001 group demonstrated significantly decreased apoptosis (p = 0.01) evident on TUNEL staining.


The administration of pan-caspase inhibitor F573 in a murine in vivo model likely mitigates liver IRI based on decreased markers of cellular injury, decreased evidence of apoptosis, and improved cytokine profiles. Anakinra with etanercept, and BMX-001 did not demonstrate convincing efficacy at reducing IRI in this model, and likely need further optimization. The positive findings set rational groundwork for future translational studies of applying F573 during normothermic ex situ liver perfusion, with the aim of improving the quality of marginal grafts.

Klíčová slova:

Aminotransferases – Apoptosis – Blood plasma – Cytokines – Histology – Ischemia – Liver transplantation – Oxidative stress


1. Peralta C, Jimenez-Castro MB, Gracia-Sancho J. Hepatic ischemia and reperfusion injury: effects on the liver sinusoidal milieu. J Hepatol. 2013;59(5):1094–106. Epub 2013/07/03. doi: 10.1016/j.jhep.2013.06.017. 23811302.

2. Linares I, Farrokhi K, Echeverri J, Kaths JM, Kollmann D, Hamar M, et al. PPAR-gamma activation is associated with reduced liver ischemia-reperfusion injury and altered tissue-resident macrophages polarization in a mouse model. PLoS One. 2018;13(4):e0195212. Epub 2018/04/05. doi: 10.1371/journal.pone.0195212. 29617419; PubMed Central PMCID: PMC5884549.

3. Cheng F, Li Y, Feng L, Li S. Effects of tetrandrine on ischemia/reperfusion injury in mouse liver. Transplant Proc. 2008;40(7):2163–6. Epub 2008/09/16. doi: 10.1016/j.transproceed.2008.07.082. 18790181.

4. Rong YP, Huang HT, Liu JS, Wei L. Protective Effects of Geniposide on Hepatic Ischemia/Reperfusion Injury. Transplant Proc. 2017;49(6):1455–60. Epub 2017/07/25. doi: 10.1016/j.transproceed.2017.02.063. 28736023.

5. Yang Y, Yang J, Jiang Q. The protective effect of huperzine A against hepatic ischemia reperfusion injury in mice. Transplant Proc. 2014;46(5):1573–7. Epub 2014/06/18. doi: 10.1016/j.transproceed.2014.01.018. 24935330.

6. Hochhauser E, Lahat E, Sultan M, Pappo O, Waldman M, Sarne Y, et al. Ultra Low Dose Delta 9-Tetrahydrocannabinol Protects Mouse Liver from Ischemia Reperfusion Injury. Cell Physiol Biochem. 2015;36(5):1971–81. Epub 2015/07/24. doi: 10.1159/000430165. 26202357.

7. Palumbo T, Nakamura K, Lassman C, Kidani Y, Bensinger SJ, Busuttil R, et al. Bruton Tyrosine Kinase Inhibition Attenuates Liver Damage in a Mouse Warm Ischemia and Reperfusion Model. Transplantation. 2017;101(2):322–31. Epub 2016/11/08. doi: 10.1097/TP.0000000000001552. 27820779; PubMed Central PMCID: PMC5263143.

8. Emamaullee JA, Davis J, Pawlick R, Toso C, Merani S, Cai SX, et al. The caspase selective inhibitor EP1013 augments human islet graft function and longevity in marginal mass islet transplantation in mice. Diabetes. 2008;57(6):1556–66. Epub 2008/03/22. doi: 10.2337/db07-1452. 18356409.

9. McCall MD, Maciver AM, Kin T, Emamaullee J, Pawlick R, Edgar R, et al. Caspase inhibitor IDN6556 facilitates marginal mass islet engraftment in a porcine islet autotransplant model. Transplantation. 2012;94(1):30–5. Epub 2012/06/19. doi: 10.1097/TP.0b013e318257745d. 22706322.

10. Pepper AR, Bruni A, Pawlick R, Wink J, Rafiei Y, Gala-Lopez B, et al. Engraftment Site and Effectiveness of the Pan-Caspase Inhibitor F573 to Improve Engraftment in Mouse and Human Islet Transplantation in Mice. Transplantation. 2017;101(10):2321–9. Epub 2017/01/11. doi: 10.1097/TP.0000000000001638. 28072753.

11. Emamaullee JA, Stanton L, Schur C, Shapiro AM. Caspase inhibitor therapy enhances marginal mass islet graft survival and preserves long-term function in islet transplantation. Diabetes. 2007;56(5):1289–98. Epub 2007/02/17. doi: 10.2337/db06-1653. 17303806.

12. McCall M, Pawlick R, Kin T, Shapiro AM. Anakinra potentiates the protective effects of etanercept in transplantation of marginal mass human islets in immunodeficient mice. Am J Transplant. 2012;12(2):322–9. Epub 2011/11/08. doi: 10.1111/j.1600-6143.2011.03796.x. 22053751.

13. Bruni A, Pepper AR, Pawlick RL, Gala-Lopez B, Gamble A, Kin T, et al. BMX-001, a novel redox-active metalloporphyrin, improves islet function and engraftment in a murine transplant model. Am J Transplant. 2018;18(8):1879–89. Epub 2018/02/22. doi: 10.1111/ajt.14705. 29464912.

14. Abe Y, Hines IN, Zibari G, Pavlick K, Gray L, Kitagawa Y, et al. Mouse model of liver ischemia and reperfusion injury: method for studying reactive oxygen and nitrogen metabolites in vivo. Free Radic Biol Med. 2009;46(1):1–7. Epub 2008/10/29. doi: 10.1016/j.freeradbiomed.2008.09.029. 18955130; PubMed Central PMCID: PMC2740994.

15. Gad SC, Sullivan DW Jr., Spasojevic I, Mujer CV, Spainhour CB, Crapo JD. Nonclinical Safety and Toxicokinetics of MnTnBuOE-2-PyP5+ (BMX-001). Int J Toxicol. 2016;35(4):438–53. Epub 2016/04/22. doi: 10.1177/1091581816642766. 27098749

16. Brockmann J, Reddy S, Coussios C, Pigott D, Guirriero D, Hughes D, et al. Normothermic perfusion: a new paradigm for organ preservation. Ann Surg. 2009;250(1):1–6. doi: 10.1097/SLA.0b013e3181a63c10. 19561463.

17. Goldaracena N, Spetzler VN, Echeverri J, Kaths JM, Cherepanov V, Persson R, et al. Inducing Hepatitis C Virus Resistance After Pig Liver Transplantation-A Proof of Concept of Liver Graft Modification Using Warm Ex Vivo Perfusion. Am J Transplant. 2017;17(4):970–8. Epub 2016/11/03. doi: 10.1111/ajt.14100. 27805315.

18. Baskin-Bey ES, Washburn K, Feng S, Oltersdorf T, Shapiro D, Huyghe M, et al. Clinical Trial of the Pan-Caspase Inhibitor, IDN-6556, in Human Liver Preservation Injury. Am J Transplant. 2007;7(1):218–25. doi: 10.1111/j.1600-6143.2006.01595.x. 17227570.

19. Abbate A, Salloum FN, Vecile E, Das A, Hoke NN, Straino S, et al. Anakinra, a recombinant human interleukin-1 receptor antagonist, inhibits apoptosis in experimental acute myocardial infarction. Circulation. 2008;117(20):2670–83. doi: 10.1161/CIRCULATIONAHA.107.740233. 18474815.

20. Goldaracena N, Echeverri J, Spetzler VN, Kaths JM, Barbas AS, Louis KS, et al. Anti-inflammatory signaling during ex vivo liver perfusion improves the preservation of pig liver grafts before transplantation. Liver Transpl. 2016;22(11):1573–83. Epub 2016/10/27. doi: 10.1002/lt.24603. 27556578.

Článek vyšel v časopise


2019 Číslo 11