Neutrophil-to-hepatocyte communication via LDLR-dependent miR-223–enriched extracellular vesicle transfer ameliorates nonalcoholic steatohepatitis
Yong He, … , George Kunos, Bin Gao
Yong He, … , George Kunos, Bin Gao
Published December 10, 2020
Citation Information: J Clin Invest. 2021;131(3):e141513. https://doi.org/10.1172/JCI141513.
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Neutrophil-to-hepatocyte communication via LDLR-dependent miR-223–enriched extracellular vesicle transfer ameliorates nonalcoholic steatohepatitis

Abstract

Neutrophil infiltration around lipotoxic hepatocytes is a hallmark of nonalcoholic steatohepatitis (NASH); however, how these 2 types of cells communicate remains obscure. We have previously demonstrated that neutrophil-specific microRNA-223 (miR-223) is elevated in hepatocytes to limit NASH progression in obese mice. Here, we demonstrated that this elevation of miR-223 in hepatocytes was due to preferential uptake of miR-223–enriched extracellular vesicles (EVs) derived from neutrophils as well other types of cells, albeit to a lesser extent. This selective uptake was dependent on the expression of low-density lipoprotein receptor (LDLR) on hepatocytes and apolipoprotein E (APOE) on neutrophil-derived EVs, which was enhanced by free fatty acids. Once internalized by hepatocytes, the EV-derived miR-223 acted to inhibit hepatic inflammatory and fibrogenic gene expression. In the absence of this LDLR- and APOE-dependent uptake of miR-223–enriched EVs, the progression of steatosis to NASH was accelerated. In contrast, augmentation of this transfer by treatment with an inhibitor of proprotein convertase subtilisin/kexin type 9, a drug used to lower blood cholesterol by upregulating LDLR, ameliorated NASH in mice. This specific role of LDLR and APOE in the selective control of miR-223–enriched EV transfer from neutrophils to hepatocytes may serve as a potential therapeutic target for NASH.

Authors

Yong He, Robim M. Rodrigues, Xiaolin Wang, Wonhyo Seo, Jing Ma, Seonghwan Hwang, Yaojie Fu, Eszter Trojnár, Csaba Mátyás, Suxian Zhao, Ruixue Ren, Dechun Feng, Pal Pacher, George Kunos, Bin Gao

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Figure 1

Immune cell–derived miR-223 is selectively transferred into the liver (hepatocytes), ameliorating NASH.

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Immune cell–derived miR-223 is selectively transferred into the liver (h...
(A) C57BL/6J mice were fed HFD or CD for 3 months. Serum and different organ samples were collected for the measurement of miR-223 levels (n = 3–4). (B–F) WT and miR-223KO mice were transplanted with WT or miR-223KO mouse bone marrow (BM). Two months later, these mice were subjected to HFD feeding for 3 months (n = 4–6). (B) Serum and liver tissue samples were collected for miR-223 measurement. (C) Frozen liver tissue sections from HFD-fed mice were analyzed by miR-223 in situ hybridization along with immunofluorescence staining of neutrophil marker MPO and cell cytoskeleton marker F-actin that was detected by using Alexa Fluor–phalloidin. Representative images of miR-223 expression (green), MPO (yellow), phalloidin (red), and nuclei (DAPI, blue) are shown. White arrows indicate MPO+ neutrophils (left panel) or miR-223+MPO+ neutrophils (right panel). Red arrows indicate miR-223+ hepatocytes. Scale bars: 10 μm. (D) Serum ALT was measured (left panel). RT-qPCR analyses of liver Ly6g and F4/80 mRNA levels (right panel). (E) Representative images of H&E staining (scale bars: 200 μm), Oil red staining (scale bars: 100 μm), Sirius red staining (scale bars: 200 μm), and Masson’s trichrome staining (scale bars: 100 μm) of liver tissue sections are shown. (F) Oil red+ area and fibrotic area per field were quantified. Values represent mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001, as determined by 2-tailed Student’s t test for comparing 2 groups (A, B, D, and F). ND, not detectable. The superscript characters shown for transplanted mice indicate the donor mouse BM.