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 6

PA induces neutrophils to release APOE-enriched EVs.

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PA induces neutrophils to release APOE-enriched EVs. Bone marrow neutrop...
Bone marrow neutrophils were cultured in serum-free medium and stimulated with vehicle or PA (0.3 mM) for 6 hours. (A) miR-223 levels in neutrophils and neutrophil-derived EVs were measured. (B) Apoe and Pu.1 mRNA levels in neutrophils were measured. (C) Western blot analyses of APOE in bone marrow neutrophils and EV marker proteins in neutrophil-derived EVs from WT and Apoe-KO mice. (D) Colocalization of APOE with an early endosome marker (EEA1), EV marker (CD63), or late endosome marker (RAB7) was determined. Representative images of these markers (red), APOE (green), and nuclei (DAPI, blue) are shown. Scale bars: 5 μm. (E) AML12 cells and primary mouse hepatocytes were incubated with DiD-labeled EVs that were derived from serum-free–cultured neutrophils. Representative images of DiD (red) and nuclei (blue) are shown, and mean fluorescence intensity per cell was quantified (bottom). Scale bars: 20 μm. Values represent means ± SEM. *P < 0.05, **P < 0.01, as determined 2-tailed Student’s t test for comparing 2 groups (A, B, and E).