Review Article                                     
Article download: 58

 

Crossmark logo
 

NLRP3 Inflammasome Activation and Pyroptotic Cell Injury in Myocardial Ischemia–Reperfusion: Molecular Mechanisms and Therapeutic Targeting

Daniel R. Whitmore MD, PhD1; Emily J. Carter FRCP2; Michael A. Reynolds MD1; Sophia L. Bennett PhD3 

1University of South Florida (USF Health), Cardiovascular Research Unit, Morsani College of Medicine, United States.
2Yale School of Medicine, Section of Cardiovascular Medicine, United States.
3Colorado State University, Department of Biomedical Sciences / Cardiovascular Research Center, United States.

DOI: 10.18081/2378-5225/11.1 
Cited by 0

 Article history: Received 12 November 2021 · Revised 03 January 2022 · Accepted 02 February 2022 · Published 22 February 2022

© 2022 Bennett et al. This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International License (CC BY 4.0). 

CC BY 4.0                                                                                                   

Abstract

Background: Myocardial ischemia–reperfusion (I/R) injury remains an important determinant of cardiomyocyte loss and impaired ventricular recovery despite successful restoration of coronary blood flow. Activation of the NLRP3 inflammasome may couple reperfusion-associated cellular stress to caspase-1-dependent gasdermin D (GSDMD) cleavage and pyroptotic cell injury. This study investigated the contribution of the NLRP3–caspase-1–GSDMD axis to myocardial I/R injury and evaluated the cardioprotective effects of pharmacological NLRP3 inhibition.

Objective: To evaluate the efficacy and safety of PARP inhibitors administered as monotherapy or in combination with androgen-receptor pathway inhibitors in patients with mCRPC.

Methods:Adult male C57BL/6J mice were randomized to Sham, Sham + MCC950, I/R, or I/R + MCC950 groups (n = 12/group). Myocardial I/R was induced by 30 min of left anterior descending coronary artery occlusion followed by 24 h of reperfusion. MCC950 (10 mg/kg, intraperitoneally) was used to inhibit NLRP3 activation. The primary outcome was infarct size relative to the area at risk (AAR), assessed by Evans blue/2,3,5-triphenyltetrazolium chloride staining. Cardiac function was evaluated by echocardiography, and myocardial injury was assessed using cardiac troponin I (cTnI), creatine kinase-MB (CK-MB), and histopathology. NLRP3, ASC, cleaved caspase-1, and GSDMD-N were evaluated as components of inflammasome/pyroptotic signaling. Myocardial IL-1β and IL-18, oxidative-stress indices, and TUNEL positivity were additionally assessed.

Results: The AAR was comparable between I/R and I/R + MCC950 groups (46.8 ± 5.7% vs 45.9 ± 5.3%; P = 0.71), whereas MCC950 reduced infarct size from 45.7 ± 6.8% to 27.4 ± 5.9% of the AAR (P < 0.001). I/R markedly impaired left ventricular systolic function, reducing left ventricular ejection fraction (LVEF) to 43.6 ± 6.1%, compared with 72.1 ± 4.3% in Sham animals; MCC950 preserved LVEF at 58.9 ± 5.4% (P < 0.001 vs I/R). MCC950 also reduced cTnI (8.74 ± 1.62 to 4.31 ± 1.18 ng/mL) and CK-MB (38.6 ± 6.7 to 21.4 ± 4.8 ng/mL; both P < 0.001) and attenuated histopathological myocardial injury. Mechanistically, I/R produced coordinated increases in NLRP3 and ASC expression, caspase-1 cleavage, GSDMD-N formation, and IL-1β and IL-18 concentrations. MCC950 significantly suppressed these changes and was additionally associated with reduced lipid peroxidation, restoration of endogenous antioxidant defenses, and decreased TUNEL-positive myocardial nuclei. NLRP3 and GSDMD-N expression correlated positively with infarct size and inversely with ventricular systolic function.

Conclusion: Myocardial I/R induces coordinated activation of the NLRP3–ASC–caspase-1–GSDMD pathway, linking inflammasome signaling to pyroptotic/inflammatory myocardial injury, infarct expansion, and acute ventricular dysfunction. Pharmacological NLRP3 inhibition attenuated downstream pyroptotic signaling and produced concordant molecular, biochemical, structural, and functional cardioprotection. These findings identify the NLRP3–caspase-1–GSDMD axis as a potentially actionable therapeutic pathway for limiting myocardial reperfusion injury.

Keywords: NLRP3 inflammasome; pyroptosis; myocardial ischemia–reperfusion injury; MCC950; gasdermin D; caspase-1; myocardial infarction.

References

  1. Yellon DM, Hausenloy DJ. Myocardial reperfusion injury. N Engl J Med. 2007;357(11):1121-1135. doi:10.1056/NEJMra071667
  2. Hausenloy DJ, Yellon DM. Myocardial ischemia-reperfusion injury: a neglected therapeutic target. J Clin Invest. 2013;123(1):92-100. doi:10.1172/JCI62874
  3. Davidson SM, Ferdinandy P, Andreadou I, et al. Multitarget strategies to reduce myocardial ischemia/reperfusion injury: JACC review topic of the week. J Am Coll Cardiol. 2019;73(1):89-99. doi:10.1016/j.jacc.2018.09.086
  4. Heusch G. Myocardial ischaemia-reperfusion injury and cardioprotection in perspective. Nat Rev Cardiol. 2020;17(12):773-789. doi:10.1038/s41569-020-0403-y
  5. Ibáñez B, Heusch G, Ovize M, Van de Werf F. Evolving therapies for myocardial ischemia/reperfusion injury. J Am Coll Cardiol. 2015;65(14):1454-1471. doi:10.1016/j.jacc.2015.02.032
  6. Frangogiannis NG. The inflammatory response in myocardial injury, repair, and remodelling. Nat Rev Cardiol. 2014;11(5):255-265. doi:10.1038/nrcardio.2014.28
  7. Prabhu SD, Frangogiannis NG. The biological basis for cardiac repair after myocardial infarction: from inflammation to fibrosis. Circ Res. 2016;119(1):91-112. doi:10.1161/CIRCRESAHA.116.303577
  8. Eltzschig HK, Eckle T. Ischemia and reperfusion—from mechanism to translation. Nat Med. 2011;17(11):1391-1401. doi:10.1038/nm.2507
  9. Kalogeris T, Baines CP, Krenz M, Korthuis RJ. Cell biology of ischemia/reperfusion injury. Int Rev Cell Mol Biol. 2012;298:229-317. doi:10.1016/B978-0-12-394309-5.00006-7
  10. Toldo S, Abbate A. The NLRP3 inflammasome in acute myocardial infarction. Nat Rev Cardiol. 2018;15(4):203-214. doi:10.1038/nrcardio.2017.161
  11. Toldo S, Mauro AG, Cutter Z, Abbate A. Inflammasome, pyroptosis, and cytokines in myocardial ischemia-reperfusion injury. Am J Physiol Heart Circ Physiol. 2018;315(6). doi:10.1152/ajpheart.00158.2018
  12. Toldo S, Marchetti C, Mauro AG, et al. Inhibition of the NLRP3 inflammasome limits the inflammatory injury following myocardial ischemia-reperfusion in the mouse. Int J Cardiol. 2016;209:215-220. doi:10.1016/j.ijcard.2016.02.043
  13. Sandanger Ø, Ranheim T, Vinge LE, et al. The NLRP3 inflammasome is up-regulated in cardiac fibroblasts and mediates myocardial ischaemia-reperfusion injury. Cardiovasc Res. 2013;99(1):164-174. doi:10.1093/cvr/cvt091
  14. Mezzaroma E, Toldo S, Farkas D, et al. The inflammasome promotes adverse cardiac remodeling following acute myocardial infarction in the mouse. Proc Natl Acad Sci U S A. 2011;108(49):19725-19730. doi:10.1073/pnas.1108586108
  15. Kawaguchi M, Takahashi M, Hata T, et al. Inflammasome activation of cardiac fibroblasts is essential for myocardial ischemia/reperfusion injury. 2011;123(6):594-604. doi:10.1161/CIRCULATIONAHA.110.982777
  16. Zuurbier CJ, Abbate A, Cabrera-Fuentes HA, et al. Innate immunity as a target for acute cardioprotection. Cardiovasc Res. 2019;115(7):1131-1142. doi:10.1093/cvr/cvy304
  17. Swanson KV, Deng M, Ting JPY. The NLRP3 inflammasome: molecular activation and regulation to therapeutics. Nat Rev Immunol. 2019;19(8):477-489. doi:10.1038/s41577-019-0165-0
  18. Kelley N, Jeltema D, Duan Y, He Y. The NLRP3 inflammasome: an overview of mechanisms of activation and regulation. Int J Mol Sci. 2019;20(13):3328. doi:10.3390/ijms20133328
  19. Broz P, Dixit VM. Inflammasomes: mechanism of assembly, regulation and signalling. Nat Rev Immunol. 2016;16(7):407-420. doi:10.1038/nri.2016.58
  20. Schroder K, Tschopp J. The inflammasomes. 2010;140(6):821-832. doi:10.1016/j.cell.2010.01.040
  21. Martinon F, Burns K, Tschopp J. The inflammasome: a molecular platform triggering activation of inflammatory caspases and processing of proIL-beta. Mol Cell. 2002;10(2):417-426. doi:10.1016/S1097-2765(02)00599-3
  22. Bauernfeind FG, Horvath G, Stutz A, et al. Cutting edge: NF-κB activating pattern recognition and cytokine receptors license NLRP3 inflammasome activation by regulating NLRP3 expression. J Immunol. 2009;183(2):787-791. doi:10.4049/jimmunol.0901363
  23. Franchi L, Muñoz-Planillo R, Núñez G. Sensing and reacting to microbes through the inflammasomes. Nat Immunol. 2012;13(4):325-332. doi:10.1038/ni.2231
  24. Guo H, Callaway JB, Ting JPY. Inflammasomes: mechanism of action, role in disease, and therapeutics. Nat Med. 2015;21(7):677-687. doi:10.1038/nm.3893
  25. He Y, Hara H, Núñez G. Mechanism and regulation of NLRP3 inflammasome activation. Trends Biochem Sci. 2016;41(12):1012-1021. doi:10.1016/j.tibs.2016.09.002
  26. Lamkanfi M, Dixit VM. Mechanisms and functions of inflammasomes. 2014;157(5):1013-1022. doi:10.1016/j.cell.2014.04.007
  27. Jo EK, Kim JK, Shin DM, Sasakawa C. Molecular mechanisms regulating NLRP3 inflammasome activation. Cell Mol Immunol. 2016;13(2):148-159. doi:10.1038/cmi.2015.95
  28. Coll RC, Robertson AAB, Chae JJ, et al. A small-molecule inhibitor of the NLRP3 inflammasome for the treatment of inflammatory diseases. Nat Med. 2015;21(3):248-255. doi:10.1038/nm.3806
  29. Coll RC, Hill JR, Day CJ, et al. MCC950 directly targets the NLRP3 ATP-hydrolysis motif for inflammasome inhibition. Nat Chem Biol. 2019;15(6):556-559. doi:10.1038/s41589-019-0277-7
  30. Tapia-Abellán A, Angosto-Bazarra D, Martínez-Banaclocha H, et al. MCC950 closes the active conformation of NLRP3 to an inactive state. Nat Chem Biol. 2019;15(6):560-564. doi:10.1038/s41589-019-0278-6
  31. Shi J, Zhao Y, Wang K, et al. Cleavage of GSDMD by inflammatory caspases determines pyroptotic cell death. 2015;526(7575):660-665. doi:10.1038/nature15514
  32. Kayagaki N, Stowe IB, Lee BL, et al. Caspase-11 cleaves gasdermin D for non-canonical inflammasome signalling. 2015;526(7575):666-671. doi:10.1038/nature15541
  33. He WT, Wan H, Hu L, et al. Gasdermin D is an executor of pyroptosis and required for interleukin-1β secretion. Cell Res. 2015;25(12):1285-1298. doi:10.1038/cr.2015.139
  34. Liu X, Zhang Z, Ruan J, et al. Inflammasome-activated gasdermin D causes pyroptosis by forming membrane pores. 2016;535(7610):153-158. doi:10.1038/nature18629
  35. Sborgi L, Rühl S, Mulvihill E, et al. GSDMD membrane pore formation constitutes the mechanism of pyroptotic cell death. EMBO J. 2016;35(16):1766-1778. doi:10.15252/embj.201694696
  36. Ding J, Wang K, Liu W, et al. Pore-forming activity and structural autoinhibition of the gasdermin family. 2016;535(7610):111-116. doi:10.1038/nature18590
  37. Aglietti RA, Estevez A, Gupta A, et al. GsdmD p30 elicited by caspase-11 during pyroptosis forms pores in membranes. Proc Natl Acad Sci U S A. 2016;113(28):7858-7863. doi:10.1073/pnas.1607769113
  38. Shi J, Gao W, Shao F. Pyroptosis: gasdermin-mediated programmed necrotic cell death. Trends Biochem Sci. 2017;42(4):245-254. doi:10.1016/j.tibs.2016.10.004
  39. Broz P, Pelegrín P, Shao F. The gasdermins, a protein family executing cell death and inflammation. Nat Rev Immunol. 2020;20(3):143-157. doi:10.1038/s41577-019-0228-2
  40. Evavold CL, Ruan J, Tan Y, Xia S, Wu H, Kagan JC. The pore-forming protein gasdermin D regulates interleukin-1 secretion from living macrophages. 2018;48(1):35-44.e6. doi:10.1016/j.immuni.2017.11.013
  41. Heilig R, Dick MS, Sborgi L, Meunier E, Hiller S, Broz P. The gasdermin-D pore acts as a conduit for IL-1β secretion in mice. Eur J Immunol. 2018;48(4):584-592. doi:10.1002/eji.201747404
  42. Xia S, Zhang Z, Magupalli VG, et al. Gasdermin D pore structure reveals preferential release of mature interleukin-1. 2021;593(7860):607-611. doi:10.1038/s41586-021-03478-3
  43. Bergsbaken T, Fink SL, Cookson BT. Pyroptosis: host cell death and inflammation. Nat Rev Microbiol. 2009;7(2):99-109. doi:10.1038/nrmicro2070
  44. Jorgensen I, Miao EA. Pyroptotic cell death defends against intracellular pathogens. Immunol Rev. 2015;265(1):130-142. doi:10.1111/imr.12287
  45. Man SM, Kanneganti TD. Regulation of inflammasome activation. Immunol Rev. 2015;265(1):6-21. doi:10.1111/imr.12296
  46. Toldo S, Mezzaroma E, Mauro AG, Salloum F, Van Tassell BW, Abbate A. The inflammasome in myocardial injury and cardiac remodeling. Antioxid Redox Signal. 2015;22(13):1146-1161. doi:10.1089/ars.2014.5989
  47. Mauro AG, Bonaventura A, Mezzaroma E, et al. NLRP3 inflammasome in acute myocardial infarction. J Cardiovasc Pharmacol. 2019;74(3):175-187.
  48. Abbate A, Toldo S, Marchetti C, Kron J, Van Tassell BW, Dinarello CA. Interleukin-1 and the inflammasome as therapeutic targets in cardiovascular disease. Circ Res. 2020;126(9):1260-1280. doi:10.1161/CIRCRESAHA.120.315937
  49. Dinarello CA. Overview of the IL-1 family in innate inflammation and acquired immunity. Immunol Rev. 2018;281(1):8-27. doi:10.1111/imr.12621
  50. Dinarello CA, Simon A, van der Meer JWM. Treating inflammation by blocking interleukin-1 in a broad spectrum of diseases. Nat Rev Drug Discov. 2012;11(8):633-652. doi:10.1038/nrd3800
  51. Ridker PM, Everett BM, Thuren T, et al. Antiinflammatory therapy with canakinumab for atherosclerotic disease. N Engl J Med. 2017;377(12):1119-1131. doi:10.1056/NEJMoa1707914
  52. Tardif JC, Kouz S, Waters DD, et al. Efficacy and safety of low-dose colchicine after myocardial infarction. N Engl J Med. 2019;381(26):2497-2505. doi:10.1056/NEJMoa1912388
  53. Nidorf SM, Fiolet ATL, Mosterd A, et al. Colchicine in patients with chronic coronary disease. N Engl J Med. 2020;383(19):1838-1847. doi:10.1056/NEJMoa2021372
  54. Ridker PM, Everett BM, Pradhan A, et al. Low-dose methotrexate for the prevention of atherosclerotic events. N Engl J Med. 2019;380(8):752-762. doi:10.1056/NEJMoa1809798
  55. Ridker PM, Lüscher TF. Anti-inflammatory therapies for cardiovascular disease. Eur Heart J. 2014;35(27):1782-1791. doi:10.1093/eurheartj/ehu203
  56. Libby P. Inflammation in atherosclerosis. 2002;420(6917):868-874. doi:10.1038/nature01323
  57. Libby P. Inflammation during the life cycle of the atherosclerotic plaque. Cardiovasc Res. 2021;117(13):2525-2536. doi:10.1093/cvr/cvab303
  58. Ross R. Atherosclerosis—an inflammatory disease. N Engl J Med. 1999;340(2):115-126. doi:10.1056/NEJM199901143400207
  59. Ong SB, Hernández-Reséndiz S, Crespo-Avilan GE, et al. Inflammation following acute myocardial infarction: multiple players, dynamic roles, and novel therapeutic opportunities. Pharmacol Ther. 2018;186:73-87. doi:10.1016/j.pharmthera.2018.01.001
  60. Frangogiannis NG. Pathophysiology of myocardial infarction. Compr Physiol. 2015;5(4):1841-1875. doi:10.1002/cphy.c150006
  61. Del Re DP, Amgalan D, Linkermann A, Liu Q, Kitsis RN. Fundamental mechanisms of regulated cell death and implications for heart disease. Physiol Rev. 2019;99(4):1765-1817. doi:10.1152/physrev.00022.2018
  62. Mishra PK, Adameova A, Hill JA, et al. Guidelines for evaluating myocardial cell death. Am J Physiol Heart Circ Physiol. 2019;317(5). doi:10.1152/ajpheart.00259.2019
  63. Davidson SM, Adameová A, Barile L, et al. Mitochondrial and mitochondrial-independent pathways of myocardial cell death during ischaemia and reperfusion injury. J Cell Mol Med. 2020;24(7):3795-3806. doi:10.1111/jcmm.15127
  64. Zhou R, Yazdi AS, Menu P, Tschopp J. A role for mitochondria in NLRP3 inflammasome activation. 2011;469(7329):221-225. doi:10.1038/nature09663
  65. Shimada K, Crother TR, Karlin J, et al. Oxidized mitochondrial DNA activates the NLRP3 inflammasome during apoptosis. 2012;36(3):401-414. doi:10.1016/j.immuni.2012.01.009
  66. Xu J, Zeng X, Niu H, et al. Normothermic ex vivo heart perfusion with NLRP3 inflammasome inhibitor MCC950 treatment improves cardiac function of circulatory death hearts after transplantation. Front Cardiovasc Med. 2023;10:1126391. doi:10.3389/fcvm.2023.1126391

BM-Publisher · Pathophysiology of Cell Injury Journal (PCIJ) · E-ISSN 2378-5225 · DOI Prefix 10.18081/pcij/2378-5225

Pathophysiology of Cell Injury Journal (PCIJ)
E-ISSN 2378-5225 · Biannual
BM-Publisher (London, UK)
Open Access

Vol 11, Issue 1 (February 2022), pp. 1–39

Download article

PDF
XML

How to cite (AMA)

Whitmore DR, Carter EJ, Reynolds MA, Bennett SL. NLRP3 Inflammasome Activation and Pyroptotic Cell Injury in Myocardial Ischemia–Reperfusion: Molecular Mechanisms and Therapeutic Targeting. Pathophysiology of Cell Injury Journal (PCIJ). 2022;11(1):1–39. doi: 10.18081/2378-5225/11.1.

More citation

(2022). . Pathophysiology of Cell Injury Journal (PCIJ), . https://pcij.net/archives/2846
. \".\" Pathophysiology of Cell Injury Journal (PCIJ), 2022, pp. . https://pcij.net/archives/2846
. . Pathophysiology of Cell Injury Journal (PCIJ). 2022;:. https://pcij.net/archives/2846
(2022) . Pathophysiology of Cell Injury Journal (PCIJ), , pp. . Available at: https://pcij.net/archives/2846
@article{2022, title = {}, journal = {Pathophysiology of Cell Injury Journal (PCIJ)}, year = {2022}, url = {https://pcij.net/archives/2846}, }
TY - JOUR TI - JO - Pathophysiology of Cell Injury Journal (PCIJ) PY - 2022 UR - https://pcij.net/archives/2846 ER -

PlumX Metrics