FLT4/VEGFR3 activates AMPK to coordinate glycometabolic reprogramming with autophagy and inflammasome activation for bacterial elimination

Li Ma; Weiyun Li; Yanbo Zhang; Linlin Qi; Qi Zhao; Na Li; Yao Lu; Luqing Zhang; Fei Zhou; Yichun Wu; Yongning He; Hongxiu Yu; Yulong He; Bin Wei; Hongyan Wang

doi:10.1080/15548627.2021.1985338

FLT4/VEGFR3 activates AMPK to coordinate glycometabolic reprogramming with autophagy and inflammasome activation for bacterial elimination

Autophagy. 2022 Jun;18(6):1385-1400. doi: 10.1080/15548627.2021.1985338. Epub 2021 Oct 10.

Authors

Li Ma¹, Weiyun Li^{1

2}, Yanbo Zhang^{1

3}, Linlin Qi⁴, Qi Zhao⁵, Na Li¹, Yao Lu¹, Luqing Zhang⁶, Fei Zhou⁶, Yichun Wu⁷, Yongning He⁷, Hongxiu Yu⁸, Yulong He⁶, Bin Wei^{4

5

9}, Hongyan Wang^{1

10

11}

Affiliations

¹ State Key Laboratory of Cell Biology, Cas Center for Excellence in Molecular Cell Science, Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, University of Chinese Academy of Sciences, Shanghai, China.
² School of Life Sciences, Xiamen University, Xiamen, Fujian Province, China.
³ Department of Immunology, Blavatnik Institute, Harvard Medical School, Boston, MA, USA.
⁴ State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, China.
⁵ School of Life Sciences, Shanghai University, Shanghai, China.
⁶ Cyrus Tang Hematology Center, Collaborative Innovation Center of Hematology, State Key Laboratory of Radiation Medicine and Protection, Cam-Su Genomic Resources Center, Soochow University, Suzhou, China.
⁷ State Key Laboratory of Molecular Biology, National Center for Protein Science Shanghai, Shanghai Science Research Center, Shanghai Key Laboratory of Molecular Andrology, Cas Center for Excellence in Molecular Cell Science, Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, University of Chinese Academy of Sciences, Shanghai, China.
⁸ Institutes of Biomedical Science, Fudan University, Shanghai, China.
⁹ School of Medicine, Cancer Center, Shanghai Tenth People's Hospital, Tongji University, Shanghai, China.
¹⁰ School of Life Science, Hangzhou Institute for Advanced Study, University of Chinese Academy of Sciences, Hangzhou, China.
¹¹ Bio-Research Innovation Center Suzhou, Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, Suzhou, Jiangsu, China.

Abstract

Macrophages rapidly undergo glycolytic reprogramming in response to macroautophagy/autophagy, inflammasome activation and pyroptosis for the clearance of bacteria. Identification the key molecules involved in these three events will provide critical potential therapeutic applications. Upon S. typhimurium infection, FLT4/VEGFR3 and its ligand VEGFC were inducibly expressed in macrophages, and FLT4 signaling inhibited CASP1 (caspase 1)-dependent inflammasome activation and pyroptosis but enhanced MAP1LC3/LC3 activation for elimination of the bacteria. Consistently, FLT4 mutants lacking the extracellular ligand-binding domain increased production of the proinflammatory metabolites such as succinate and lactate, and reduced antimicrobial metabolites including citrate and NAD(P)H in macrophages and liver upon infection. Mechanistically, FLT4 recruited AMP-activated protein kinase (AMPK) and phosphorylated Y247 and Y441/442 in the PRKAA/alpha subunit for AMPK activation. The AMPK agonist AICAR could rescue glycolytic reprogramming and inflammasome activation in macrophages expressing the mutant FLT4, which has potential translational application in patients carrying Flt4 mutations to prevent recurrent infections. Collectively, we have elucidated that the FLT4-AMPK module in macrophages coordinates glycolytic reprogramming, autophagy, inflammasome activation and pyroptosis to eliminate invading bacteria.Abbreviations: 3-MA: 3-methyladenine; AICAR: 5-aminoimidazole-4-carboxamide1-β-D-ribofuranoside; AMP: adenosine monophosphate; AMPK: AMP-activated protein kinase; ATP: adenosine triphosphate; BMDM: bone marrow-derived macrophage; CASP1: caspase 1; CFUs: colony-forming units; FLT4/VEGFR3: FMS-like tyrosine kinase 4; GFP: green fluorescent protein; LDH: lactate dehydrogenase; LPS: lipopolysaccharide; MAP1LC3/LC3: microtubule-associated protein 1 light chain 3; PEM: peritoneal exudate macrophage; PRKAA1/AMPKα1: protein kinase, AMP-activated, alpha 1 catalytic subunit; PYCARD/ASC: PYD and CARD domain containing; ROS: reactive oxygen species; SQSTM1/p62: sequestosome 1; TLR4: toll-like receptor 4; ULK1: unc-51 like autophagy activating kinase 1; VEGFC: vascular endothelial growth factor C; WT: wild type.

Keywords: AMPK; FLT4/VEGFR3; glycolytic reprogramming; inflammasome; pyroptosis.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

AMP-Activated Protein Kinases / metabolism
Adenosine Monophosphate
Autophagy* / physiology
Bacteria / metabolism
Caspase 1
Humans
Inflammasomes* / metabolism
Ligands
Lipopolysaccharides
Vascular Endothelial Growth Factor C
Vascular Endothelial Growth Factor Receptor-3

Substances

Inflammasomes
Ligands
Lipopolysaccharides
Vascular Endothelial Growth Factor C
Adenosine Monophosphate
FLT4 protein, human
Vascular Endothelial Growth Factor Receptor-3
AMP-Activated Protein Kinases
Caspase 1

Grants and funding

This work was supported by grants from the Ministry of Science and Technology of the People’s Republic of China (2016YFD0500407, 2016YFC0905902, 2018YFA0800702, 2016YFD0500207, 2016YFC0905900), National Natural Science Foundation of China (81930038, 81825011, 81630043, 81671572, 81571552, 81701569, 31700781, 81801575, 30671038, 81961160738), the Strategic Priority Research Program of the Chinese Academy of Sciences (XDB19000000), and the Priority Academic Program Development of Jiangsu Higher Education Institutions. Dr. Hongyan Wang is supported by the Hundred Talents Program of the Chinese Academy of Sciences.