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Adipose tissue inflammation and systemic insulin resistance in mice with diet-induced obesity is possibly associated with disruption of PFKFB3 in hematopoietic cells

Abstract

Obesity-associated inflammation in white adipose tissue (WAT) is a causal factor of systemic insulin resistance; however, precisely how immune cells regulate WAT inflammation in relation to systemic insulin resistance remains to be elucidated. The present study examined a role for 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase 3 (PFKFB3) in hematopoietic cells in regulating WAT inflammation and systemic insulin sensitivity. Male C57BL/6J mice were fed a high-fat diet (HFD) or low-fat diet (LFD) for 12 weeks and examined for WAT inducible 6-phosphofructo-2-kinase (iPFK2) content, while additional HFD-fed mice were treated with rosiglitazone and examined for PFKFB3 mRNAs in WAT stromal vascular cells (SVC). Also, chimeric mice in which PFKFB3 was disrupted only in hematopoietic cells and control chimeric mice were also fed an HFD and examined for HFD-induced WAT inflammation and systemic insulin resistance. In vitro, adipocytes were co-cultured with bone marrow-derived macrophages and examined for adipocyte proinflammatory responses and insulin signaling. Compared with their respective levels in controls, WAT iPFK2 amount in HFD-fed mice and WAT SVC PFKFB3 mRNAs in rosiglitazone-treated mice were significantly increased. When the inflammatory responses were analyzed, peritoneal macrophages from PFKFB3-disrputed mice revealed increased proinflammatory activation and decreased anti-inflammatory activation compared with control macrophages. At the whole animal level, hematopoietic cell-specific PFKFB3 disruption enhanced the effects of HFD feeding on promoting WAT inflammation, impairing WAT insulin signaling, and increasing systemic insulin resistance. In vitro, adipocytes co-cultured with PFKFB3-disrupted macrophages revealed increased proinflammatory responses and decreased insulin signaling compared with adipocytes co-cultured with control macrophages. These results suggest that PFKFB3 disruption in hematopoietic cells only exacerbates HFD-induced WAT inflammation and systemic insulin resistance.

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Fig. 1: PFKFB3 expression is relevant to adipose tissue inflammation in mice with diet-induced obesity.
Fig. 2: Disruption of PFKFB3 enhances macrophage proinflammatory activation and impairs macrophage anti-inflammatory activation.
Fig. 3: PFKFB3 regulation of macrophage activation at the level of transcriptome.
Fig. 4: Disruption of PFKFB3 in hematopoietic cells does not alter diet-induced adiposity in mice.
Fig. 5: Disruption of PFKFB3 in hematopoietic cells exacerbates diet-induced insulin resistance and glucose intolerance.
Fig. 6: Disruption of PFKFB3 in hematopoietic cells exacerbates diet-induced WAT inflammation and decreases WAT insulin signaling.
Fig. 7: PFKFB3-disrupted macrophages enhance adipocyte proinflammatory responses and decrease adipocyte insulin signaling.

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Acknowledgements

This work was supported in part by the Hickam Endowed Chair, Gastroenterology, Medicine, Indiana University and the Indiana University Health – Indiana University School of Medicine Strategic Research Initiative (GA, HF, FM), the Senior Research Career Scientist to GA (5I01BX000574) and VA Merit Award and Research Career Scientist to HF (1I01BX003031) from the United States Department of Veteran’s Affairs, Biomedical Laboratory Research and Development Service, and NIH grants DK076898, and DK110035 to GA and SG and DK108959 and DK119421 to HF, Indianapolis, Indiana. In addition, this work was supported in whole or in part by grants from the American Diabetes Association (1-10-BS-76 to CW) and NIH (DK095828 and DK124854 to CW). CW is also supported by the Hatch Program of the National Institutes of Food and Agriculture (NIFA). XG is supported by Fundamental Research Funds of Shandong University (Grant No. 2017TB0028), Young Scholars Program of Shandong University (Grant No. 2018WLJH33), and National Natural Science Foundation of China (Grant No. 81803224). The views expressed in this article are those of the authors and do not necessarily represent the views of the Department of Veterans Affairs.

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BZ, XG, HX and HL carried out most of experiments involving mice. BZ and HL carried out most of the experiments involving cells. BZ, XG, HX, HL, and TZ collected tissue and cell samples and performed molecular and biochemical assays. HL performed histological assays. JC performed bioinformatic analysis. BJ, YW, QY, JC, SG, FM, HF, and GA contributed to scientific discussion. CW came up the concept of the study and supervised all experiments and wrote the paper.

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Correspondence to Chaodong Wu.

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Zhu, B., Guo, X., Xu, H. et al. Adipose tissue inflammation and systemic insulin resistance in mice with diet-induced obesity is possibly associated with disruption of PFKFB3 in hematopoietic cells. Lab Invest 101, 328–340 (2021). https://doi.org/10.1038/s41374-020-00523-z

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