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Adipocyte and Cell Biology

The endoplasmic reticulum stress protein GRP94 modulates cathepsin L activity in M2 macrophages in conditions of obesity-associated inflammation and contributes to their pro-inflammatory profile

International Journal of Obesity (2024)Cite this article

Subjects

Abstract

Background/objectives

Adipose tissue macrophages (ATM) are key actors in the pathophysiology of obesity-related diseases. They have a unique intermediate M2-M1 phenotype which has been linked to endoplasmic reticulum (ER) stress. We previously reported that human M2 macrophages treated with the ER stress inducer thapsigargin switched to a pro-inflammatory phenotype that depended on the stress protein GRP94. In these conditions, GRP94 promoted cathepsin L secretion and was co-secreted with complement C3. As cathepsin L and complement C3 have been reported to play a role in the pathophysiology of obesity, in this work we studied the involvement of GRP94 in the pro-inflammatory phenotype of ATM.

Methods

GRP94, cathepsin L and C3 expression were analyzed in CD206 + ATM from mice, WT or obesity-resistant transgenic fat-1, fed a high-fat diet (HFD) or a standard diet. GRP94 colocalization with cathepsin L and C3 and its effects were analyzed in human primary macrophages using thapsigargin as a control to induce ER stress and palmitic acid (PA) as a driver of metabolic activation.

Results

In WT, but not in fat-1 mice, fed a HFD, we observed an increase in crown-like structures consisting of CD206 + pSTAT1+ macrophages showing high expression of GRP94 that colocalized with cathepsin L and C3. In vitro experiments showed that PA favored a M2-M1 switch depending on GRP94. This switch was prevented by omega-3 fatty acids. PA-induced GRP94-cathepsin L colocalization and a decrease in cathepsin L enzymatic activity within the cells (while the enzymatic activity in the extracellular medium was increased). These effects were prevented by the GRP94 inhibitor PU-WS13.

Conclusions

GRP94 is overexpressed in macrophages both in in vivo and in vitro conditions of obesity-associated inflammation and is involved in changing their profile towards a more pro-inflammatory profile. It colocalizes with complement C3 and cathepsin L and modulates cathepsin L activity.

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Fig. 1: CD206+ macrophages in crown-like structures in adipose tissue of obese mice are pSTAT1 + and highly expressed GRP94.
Fig. 2: CD206+GRP94+pSTAT1+ macrophages in CLS highly express complement C3, C3aR and cathepsin L.
Fig. 3: Palmitic acid induces a switch to a more pro-inflammatory profile in human primary M2 macrophages, which depends on the UPR IRE1α and PERK pathways.
Fig. 4: The pro-inflammatory switch induced by PA treatment in M2 macrophages is dependent on GRP94.
Fig. 5: GRP94 colocalizes with complement C3 and cathepsin L in PA-treated M2 macrophages.
Fig. 6: GRP94 modulates cathepsin L after PA or Tg treatment.

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Data availability

The data generated during and/or analyzed during the current study are available from the corresponding author upon reasonable request.

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Acknowledgements

We thank Nicolas Pernet from the uB Cytometry platform for helping in FACS analysis. This work was supported by grants from the Agence National de la Recherche, Institut Nationale contre le Cancer, Ligue Nationale Contre le Cancer (“Labelled team” to CG) and the Conseil Régional de Bourgogne. The work was also supported by a French Government grant managed by the French National Research Agency under the program ISITE-BFC and LabEX LipSTIC, with reference ANR-11-LABX-0021. We thank the European Union program FEDER for their financial support. FW has a fellowship from ISITE-BFC (Investissements d’Avenir” program, contract ANR-15-IDEX-0003) and from Ningbo University (CHINA).

Author information

Author notes

  1. These authors contributed equally: Fangmin Wang, Valentin Baverel.

Authors and Affiliations

  1. UFR des Sciences de Santé, Université de Bourgogne, Dijon, France

    Fangmin Wang, Valentin Baverel, Killian Chaumonnot, Carmen Garrido & Evelyne Kohli

  2. UMR INSERM/uB/AGROSUP 1231, Team HSP-Pathies, labellisée Ligue Nationale contre le Cancer and Laboratoire d’Excellence LipSTIC, UBFC, Dijon, France

    Fangmin Wang, Valentin Baverel, Killian Chaumonnot, Carmen Garrido & Evelyne Kohli

  3. Zhejiang Provincial Key Lab of Addiction, The Affiliated Kangning Hospital of Ningbo University, Ningbo Kangning Hospital, Ningbo University, Ningbo, China

    Fangmin Wang & Wenhua Zhou

  4. UMR INSERM/uB/AGROSUP 1231, Team Lipness, Labellisée Laboratoire d’Excellence LipSTIC, Dijon, France

    Amina Bourragat, Jerome Bellenger, Sandrine Bellenger & Michel Narce

  5. UFR Sciences Vie Terre Environnement, Université de Bourgogne, Dijon, France

    Amina Bourragat, Jerome Bellenger, Sandrine Bellenger & Michel Narce

  6. Centre Anti-Cancéreux Georges François Leclerc, Dijon, France

    Carmen Garrido

  7. CHU, Dijon, France

    Evelyne Kohli

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Contributions

Conceptualization: EK. Methodology: FW, VB, KC, AB, SB, JB. Data Analysis: FW, VB, EK. Original Draft Preparation: FW, VB Review & Editing: EK, CG, JB, WZ and MN. Supervision: EK. Funding acquisition: CG and MN. EK is the guarantor of this work and, as such, had full access to all the data in the study and takes responsibility for the integrity of the data and the accuracy of the data analysis.

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Correspondence to Evelyne Kohli.

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Wang, F., Baverel, V., Chaumonnot, K. et al. The endoplasmic reticulum stress protein GRP94 modulates cathepsin L activity in M2 macrophages in conditions of obesity-associated inflammation and contributes to their pro-inflammatory profile. Int J Obes (2024). https://doi.org/10.1038/s41366-024-01478-7

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