Pharmacological Inhibition of N-Acylethanolamine Acid Amidase (NAAA) Mitigates Intestinal Fibrosis Through Modulation of Macrophage Activity

Overview

Journal J Crohns Colitis

Publisher Oxford University Press

Date 2024 Aug 30

PMID 39211986

Authors

Maria Francesca Nani

Ester Pagano

Paola De Cicco

Giuseppe Lucariello

Fabio Cattaneo

Francesca Paola Tropeano

Donatella Cicia

Rebecca Amico

Federica Raucci

Giuseppe Ercolano

Francesco Maione

Maria Michela Rinaldi

Fabiana Esposito

Rosario Ammendola

Gaetano Luglio

Raffaele Capasso

Alexandros Makriyannis

Stefania Petrosino

Francesca Borrelli

Barbara Romano

Angelo A Izzo

Affiliations

Soon will be listed here.

Abstract

Background And Aims: Intestinal fibrosis, a frequent complication of inflammatory bowel disease, is characterized by stricture formation with no pharmacological treatment to date. N-acylethanolamine acid amidase (NAAA) is responsible for the hydrolysis of acylethanolamides (AEs, eg, palmitoylethanolamide and oleoylethanolamide). Here, we investigated NAAA and AE signaling in gut fibrosis.

Methods: NAAA and AE signaling were evaluated in human intestinal specimens from patients with stenotic Crohn's disease (CD). Gut fibrosis was induced by 2,4,6-trinitrobenzenesulfonic acid, monitored by colonoscopy, and assessed by qRT-PCR, histological analyses, and confocal microscopy. Immune cells in mesenteric lymph nodes were analyzed by FACS. Colonic fibroblasts were cultured in conditioned media derived from polarized or non-polarized bone marrow-derived macrophages (BMDMs). IL-23 signaling was evaluated by qRT-PCR, ELISA, FACS, and western blot in BMDMs and in lamina propria CX3CR1+ cells.

Results: In ileocolonic human CD strictures, increased transcript expression of NAAA was observed with a decrease in its substrates oleoylethanolamide and palmitoylethanolamide. NAAA inhibition reduced intestinal fibrosis in vivo, as indicated by a decrease in inflammatory parameters, collagen deposition, and fibrosis-related genes, including those involved in epithelial-to-mesenchymal transition. More in-depth studies revealed modulation of the immune response related to IL-23 following NAAA inhibition. The antifibrotic actions of NAAA inhibition are mediated by Mφ and M2 macrophages that indirectly affect fibroblast collagenogenesis. NAAA inhibitor AM9053 normalized IL-23 signaling in BMDMs and in lamina propria CX3CR1+ cells.

Conclusions: Our findings provide new insights into the pathophysiological mechanism of intestinal fibrosis and identify NAAA as a promising target for the development of therapeutic treatments to alleviate CD-related fibrosis.

References

West J, Zvonok N, Whitten K, Wood J, Makriyannis A . Mass spectrometric characterization of human N-acylethanolamine-hydrolyzing acid amidase. J Proteome Res. 2011; 11(2):972-81. PMC: 3706083. DOI: 10.1021/pr200735a. View

Alhouayek M, Bottemanne P, Makriyannis A, Muccioli G . N-acylethanolamine-hydrolyzing acid amidase and fatty acid amide hydrolase inhibition differentially affect N-acylethanolamine levels and macrophage activation. Biochim Biophys Acta Mol Cell Biol Lipids. 2017; 1862(5):474-484. DOI: 10.1016/j.bbalip.2017.01.001. View

Honzawa Y, Nakase H, Shiokawa M, Yoshino T, Imaeda H, Matsuura M . Involvement of interleukin-17A-induced expression of heat shock protein 47 in intestinal fibrosis in Crohn's disease. Gut. 2014; 63(12):1902-12. DOI: 10.1136/gutjnl-2013-305632. View

Chen L, Li L, Chen J, Li L, Zheng Z, Ren J . Oleoylethanolamide, an endogenous PPAR-α ligand, attenuates liver fibrosis targeting hepatic stellate cells. Oncotarget. 2016; 6(40):42530-40. PMC: 4767450. DOI: 10.18632/oncotarget.6466. View

de Lange K, Moutsianas L, Lee J, Lamb C, Luo Y, Kennedy N . Genome-wide association study implicates immune activation of multiple integrin genes in inflammatory bowel disease. Nat Genet. 2017; 49(2):256-261. PMC: 5289481. DOI: 10.1038/ng.3760. View

Pryimak N, Zaiachuk M, Kovalchuk O, Kovalchuk I . The Potential Use of Cannabis in Tissue Fibrosis. Front Cell Dev Biol. 2021; 9:715380. PMC: 8542845. DOI: 10.3389/fcell.2021.715380. View

Scharl M, Huber N, Lang S, Furst A, Jehle E, Rogler G . Hallmarks of epithelial to mesenchymal transition are detectable in Crohn's disease associated intestinal fibrosis. Clin Transl Med. 2015; 4:1. PMC: 4384762. DOI: 10.1186/s40169-015-0046-5. View

Rieder F, Fiocchi C . Intestinal fibrosis in IBD--a dynamic, multifactorial process. Nat Rev Gastroenterol Hepatol. 2009; 6(4):228-35. DOI: 10.1038/nrgastro.2009.31. View

Vellecco V, Saviano A, Raucci F, Casillo G, Mansour A, Panza E . Interleukin-17 (IL-17) triggers systemic inflammation, peripheral vascular dysfunction, and related prothrombotic state in a mouse model of Alzheimer's disease. Pharmacol Res. 2022; 187:106595. DOI: 10.1016/j.phrs.2022.106595. View

10.

Garrido-Trigo A, Corraliza A, Veny M, Dotti I, Melon-Ardanaz E, Rill A . Macrophage and neutrophil heterogeneity at single-cell spatial resolution in human inflammatory bowel disease. Nat Commun. 2023; 14(1):4506. PMC: 10372067. DOI: 10.1038/s41467-023-40156-6. View

11.

DAlessio S, Ungaro F, Noviello D, Lovisa S, Peyrin-Biroulet L, Danese S . Revisiting fibrosis in inflammatory bowel disease: the gut thickens. Nat Rev Gastroenterol Hepatol. 2021; 19(3):169-184. DOI: 10.1038/s41575-021-00543-0. View

12.

Setten E, Castagna A, Nava-Sedeno J, Weber J, Carriero R, Reppas A . Understanding fibrosis pathogenesis via modeling macrophage-fibroblast interplay in immune-metabolic context. Nat Commun. 2022; 13(1):6499. PMC: 9618579. DOI: 10.1038/s41467-022-34241-5. View

13.

Fusco R, Cordaro M, Genovese T, Impellizzeri D, Siracusa R, Gugliandolo E . Adelmidrol: A New Promising Antioxidant and Anti-Inflammatory Therapeutic Tool in Pulmonary Fibrosis. Antioxidants (Basel). 2020; 9(7). PMC: 7402091. DOI: 10.3390/antiox9070601. View

14.

Wang J, Zheng J, Kulkarni A, Wang W, Garg S, Prather P . Palmitoylethanolamide regulates development of intestinal radiation injury in a mast cell-dependent manner. Dig Dis Sci. 2014; 59(11):2693-703. PMC: 4213290. DOI: 10.1007/s10620-014-3212-5. View

15.

Bode J, Ehlting C, Haussinger D . The macrophage response towards LPS and its control through the p38(MAPK)-STAT3 axis. Cell Signal. 2012; 24(6):1185-94. DOI: 10.1016/j.cellsig.2012.01.018. View

16.

Dohi T, Fujihashi K, Kiyono H, Elson C, McGhee J . Mice deficient in Th1- and Th2-type cytokines develop distinct forms of hapten-induced colitis. Gastroenterology. 2000; 119(3):724-33. DOI: 10.1053/gast.2000.16500. View

17.

Liang Z, Tang Z, Zhu C, Li F, Chen S, Han X . Intestinal CXCR6 ILC3s migrate to the kidney and exacerbate renal fibrosis via IL-23 receptor signaling enhanced by PD-1 expression. Immunity. 2024; 57(6):1306-1323.e8. PMC: 11539045. DOI: 10.1016/j.immuni.2024.05.004. View

18.

Hwang S, Chung K . Targeting fatty acid metabolism for fibrotic disorders. Arch Pharm Res. 2021; 44(9-10):839-856. DOI: 10.1007/s12272-021-01352-4. View

19.

Ribeiro A, Pontis S, Mengatto L, Armirotti A, Chiurchiu V, Capurro V . A Potent Systemically Active N-Acylethanolamine Acid Amidase Inhibitor that Suppresses Inflammation and Human Macrophage Activation. ACS Chem Biol. 2015; 10(8):1838-46. PMC: 4546552. DOI: 10.1021/acschembio.5b00114. View

20.

Palumbo-Zerr K, Horn A, Distler A, Zerr P, Dees C, Beyer C . Inactivation of fatty acid amide hydrolase exacerbates experimental fibrosis by enhanced endocannabinoid-mediated activation of CB1. Ann Rheum Dis. 2012; 71(12):2051-4. DOI: 10.1136/annrheumdis-2012-201823. View