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Endoplasmic reticulum stress is attenuated by glycolysis in lymphatic malformations

Pediatric Research (2024)Cite this article

Abstract

Background

This study aims to investigate the role of endoplasmic reticulum stress (ER stress) in human dermal lymphatic endothelial cells (HDLECs) and lymphatic malformations (LMs) and its relationship with aerobic glycolysis and inflammation.

Methods

The proliferation and apoptosis of HDLECs were examined with lipopolysaccharide (LPS) treatment. ER stress-associated proteins and glycolysis-related markers were detected by western blot. Glycolysis indexes were detected by seahorse analysis and lactic acid production assay kits. Immunohistochemistry was used to reveal the ER stress state of lymphatic endothelial cells (LECs) in LMs.

Results

LPS induced ER stress in HDLECs but did not trigger detectable apoptosis. Intriguingly, LPS-treated HDLECs also showed increased glycolysis flux. Knockdown of Hexokinase 2, a key enzyme for aerobic glycolysis, significantly inhibited the ability of HDLECs to resist ER stress-induced apoptosis. Moreover, compared to normal skin, glucose-regulated protein 78 (GRP78/BIP), and phosphorylation protein kinase R-like kinase (p-PERK), two key ER stress-associated markers, were upregulated in LECs of LMs, which was correlated with the inflected state. In addition, excessively activated ER stress inhibited the progression of LMs in rat models.

Conclusions

These data indicate that glycolysis could rescue activated ER stress in HDLECs, which is required for the accelerated development of LMs.

Impact

  • Inflammation enhances both ER stress and glycolysis in LECs while glycolysis is required to attenuate the pro-apoptotic effect of ER stress.

  • Endoplasmic reticulum (ER) stress is activated in lymphatic endothelial cells (LECs) of LMs, especially in inflammatory condition.

  • The expression of ER stress-related proteins is increased in LMs and correlated with Hexokinase 2 expression.

  • Pharmacological activation of ER stress suppresses the formation of LM lesions in the rat model.

  • ER stress may be a promising and effective therapeutic target for the treatment of LMs.

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Fig. 1: Effects of LPS on ER stress, proliferation, and apoptosis in HDLECs.
Fig. 2: LPS enhances the aerobic glycolysis in HDLECs.
Fig. 3: HK2 knockdown raises HDLECs sensitivity to ER stress-induced apoptosis.
Fig. 4: Co-expression of GRP78 (BIP), p-PERK and HK2 in LM LECs.
Fig. 5: Tunicamycin-induced ER stress suppresses the formation of LM lesions in Rat Models.
Fig. 6: Immunohistochemical staining of ki-67 and TUNEL staining in LM lesions of rat models.

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Data availability

The datasets used and/or analyses during the current study are available from the corresponding author on reasonable request.

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Funding

This study was funded by the National Natural Science Foundation of China (NSFC) grant 82370978, 81741082 (Y.C.), grant 81800994 (W.Z.), grant 81972548 (C.H.), National Health Commission of the People’s Republic of China grant 2019ZX09302011 (D.M. and Y.C.), Wuhan Science and Technology Bureau grant 2020020601012212 (Y. C.).

Author information

Author notes

  1. These authors contributed equally: Xuecong Li, Wenqun Zhong, Hao Jiang.

Authors and Affiliations

  1. State Key Laboratory of Oral & Maxillofacial Reconstruction and Regeneration, Key Laboratory of Oral Biomedicine Ministry of Education, Hubei Key Laboratory of Stomatology, School & Hospital of Stomatology, Wuhan University, Wuhan, China

    Xuecong Li, Wenqun Zhong, Hao Jiang, Peipei Wang, Maosheng Chai, Tianshuang Zhu, Jingjing Liu, Congfa Huang, Shaodong Yang & Yu Cai

  2. Department of Oral & Maxillofacial Surgery, School & Hospital of Stomatology, Wuhan University, Wuhan, China

    Xuecong Li, Wenqun Zhong, Hao Jiang, Peipei Wang, Maosheng Chai, Tianshuang Zhu, Jingjing Liu, Congfa Huang & Yu Cai

  3. Department of Pathology, School & Hospital of Stomatology, Wuhan University, Wuhan, China

    Shaodong Yang

  4. Wuhan Conform Pharm Co., Ltd, Wuhan, China

    Dongsheng Mou & Xiaoshun Zhou

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  1. Xuecong Li
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Contributions

W. Z., X. L. and H. J.: acquisition of data, analysis and interpretation of data; P. W.: interpretation of data; M. C., T. Z. and J. L.: analysis and interpretation of data. S. Y., D. M. and X. Z.: acquisition of data. C. H.: revising it critically for important intellectual conten; W. Z. and Y. C.: conception and design interpretation of data, revising it critically for important intellectual content and final approval of the version to be published.

Corresponding author

Correspondence to Yu Cai.

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Li, X., Zhong, W., Jiang, H. et al. Endoplasmic reticulum stress is attenuated by glycolysis in lymphatic malformations. Pediatr Res (2024). https://doi.org/10.1038/s41390-024-03181-9

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