CCL24 regulates biliary inflammation and fibrosis in primary sclerosing cholangitis
Raanan Greenman, … , Amnon Peled, Adi Mor
Raanan Greenman, … , Amnon Peled, Adi Mor
Published June 22, 2023
Citation Information: JCI Insight. 2023;8(12):e162270. https://doi.org/10.1172/jci.insight.162270.
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Research Article Hepatology Inflammation

CCL24 regulates biliary inflammation and fibrosis in primary sclerosing cholangitis

Abstract

ˆCCL24 is a pro-fibrotic, pro-inflammatory chemokine expressed in several chronic fibrotic diseases. In the liver, CCL24 plays a role in fibrosis and inflammation, and blocking CCL24 led to reduced liver injury in experimental models. We studied the role of CCL24 in primary sclerosing cholangitis (PSC) and evaluated the potential therapeutic effect of blocking CCL24 in this disease. Multidrug resistance gene 2–knockout (Mdr2–/–) mice demonstrated CCL24 expression in liver macrophages and were used as a relevant experimental PSC model. CCL24-neutralizing monoclonal antibody, CM-101, significantly improved inflammation, fibrosis, and cholestasis-related markers in the biliary area. Moreover, using spatial transcriptomics, we observed reduced proliferation and senescence of cholangiocytes following CCL24 neutralization. Next, we demonstrated that CCL24 expression was elevated under pro-fibrotic conditions in primary human cholangiocytes and macrophages, and it induced proliferation of primary human hepatic stellate cells and cholangiocytes, which was attenuated following CCL24 inhibition. Correspondingly, CCL24 was found to be highly expressed in liver biopsies of patients with PSC. CCL24 serum levels correlated with Enhanced Liver Fibrosis score, most notably in patients with high alkaline phosphatase levels. These results suggest that blocking CCL24 may have a therapeutic effect in patients with PSC by reducing liver inflammation, fibrosis, and cholestasis.

Authors

Raanan Greenman, Michal Segal-Salto, Neta Barashi, Ophir Hay, Avi Katav, Omer Levi, Ilan Vaknin, Revital Aricha, Sarit Aharoni, Tom Snir, Inbal Mishalian, Devorah Olam, Johnny Amer, Ahmad Salhab, Rifaat Safadi, Yaakov Maor, Palak Trivedi, Christopher J. Weston, Francesca Saffioti, Andrew Hall, Massimo Pinzani, Douglas Thorburn, Amnon Peled, Adi Mor

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Figure 5

CCL24 is upregulated under pro-fibrotic conditions in human macrophages, hepatic stellate cells, and cholangiocytes.

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CCL24 is upregulated under pro-fibrotic conditions in human macrophages,...
(A) M2 macrophages upregulate CCL24 gene expression (n = 3–4). (B) Secreted levels of CCL24 expression measured in M2 macrophages incubated with or without HSCs (n = 6). (C) Gene expression of CCL24 in HSCs cultured with M0 or M2 macrophage conditioned medium (M0-CM and M2-CM, respectively) (n = 4). (D) Gene expression of CCL24 in LX2 cells cultured with starvation medium or M0 or M2 macrophage conditioned medium (M0-CM and M2-CM, respectively). IL-4 was supplemented at 20 ng/mL (n = 3). (E) CCL24 gene expression in primary cholangiocytes following treatment with IL-4 or IL-13 (n = 3–6). (F) CCL24 quantified by ELISA following different treatments of primary cholangiocytes (n = 3). (G) Secreted levels of CCL24 in macrophages (M0) cocultured with cholangiocytes that had been pretreated with vehicle, IL-4 or IL-13. Upper scheme, experiment procedure: cholangiocytes were treated for 3 days with vehicle, IL-4 or IL-13, then washed, and M0 cells were added on top of the cholangiocytes for 24 hours. Lower graph, detection by ELISA (n = 7). All data are presented as mean ± SEM. *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, ****P ≤ 0.0001, t test (A–C) or ANOVA followed by Holm-Šídák multiple comparisons test (D–F).