ABSTRACT Chronic inflammation is a common feature of obesity, with elevated cytokines such as interleukin-1 (IL-1) in the circulation and tissues. Here, we report an unconventional

IL-1R–MyD88–IRAK2–PHB/OPA1 signaling axis that reprograms mitochondrial metabolism in adipocytes to exacerbate obesity. IL-1 induced recruitment of IRAK2 Myddosome to mitochondria outer

membranes via recognition by TOM20, followed by TIMM50-guided translocation of IRAK2 into mitochondria inner membranes, to suppress oxidative phosphorylation and fatty acid oxidation,

thereby attenuating energy expenditure. Adipocyte-specific MyD88 or IRAK2 deficiency reduced high-fat-diet-induced weight gain, increased energy expenditure and ameliorated insulin

critical link between inflammation and metabolism, representing a novel therapeutic target for patients with obesity. Access through your institution Buy or subscribe This is a preview of

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* Log in * Learn about institutional subscriptions * Read our FAQs * Contact customer support SIMILAR CONTENT BEING VIEWED BY OTHERS IL-1R-IRAKM-SLC25A1 SIGNALING AXIS REPROGRAMS LIPOGENESIS

IN ADIPOCYTES TO PROMOTE DIET-INDUCED OBESITY IN MICE Article Open access 18 May 2022 FADD REGULATES ADIPOSE INFLAMMATION, ADIPOGENESIS, AND ADIPOCYTE SURVIVAL Article Open access 15 July

2024 ACTIVATION OF THE ADIPOCYTE CREB/CRTC PATHWAY IN OBESITY Article Open access 22 October 2021 DATA AVAILABILITY The primary data for analysis of all figures and supplementary figures are

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Scholar  Download references ACKNOWLEDGEMENTS This work was supported by grants from the NIH (2P01HL029582, R01 AA023722, P01CA062220, R01 HL122283 and P50AA024333) and National Multiple

Sclerosis Society (RG5130A2/1). H.Z. was supported by a Postdoctoral Research Fellowship Award (1-16-PDF-138) from the American Diabetes Association. AUTHOR INFORMATION Author notes * Julie

A. Carman Present address: Immunology Discovery, Janssen Research and Development, Spring House, PA, USA * These authors contributed equally: Hao Zhou, Han Wang, Minjia Yu. AUTHORS AND

AFFILIATIONS * Department of Inflammation and Immunity, Lerner Research Institute, Cleveland Clinic, Cleveland, OH, USA Hao Zhou, Han Wang, Minjia Yu, Wen Qian, Fangqiang Tang, Weiwei Liu, 

Hui Yang, Ruth E. McDowell, Junjie Zhao & Xiaoxia Li * School of Life Sciences, Lanzhou University, Lanzhou, China Han Wang * Department of Medicine, Mount Auburn Hospital, Harvard

Medical School, Cambridge, MA, USA Minjia Yu * Department of Cardiovascular and Metabolic Sciences, Lerner Research Institute, Cleveland Clinic, Cleveland, OH, USA Rebecca C. Schugar, 

Christopher Hine, Jonathan D. Smith, Paul L. Fox & J. Mark Brown * Discovery Biology, Bristol Myers Squibb, Princeton, NJ, USA Ji Gao, Ashok Dongre & Julie A. Carman * Imaging Core,

Lerner Research Institute, Cleveland Clinic, Cleveland, OH, USA Mei Yin & Judith A. Drazba * University of Ottawa and Ottawa Hospital, Ottawa, Ontario, Canada Robert Dent * Department of

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publications You can also search for this author inPubMed Google Scholar CONTRIBUTIONS H.Z., H.W., M. Yu, R.C.S., W.Q., F.T., W.L., H.Y., R.E.M. and J.Z. conducted the experiments. J.A.C.,

J.G. and A.D. performed the proteomics analysis. R.D. collected human adipose tissue samples. M. Yin and J.A.D. performed the electron microscopy analysis. H.Z. and X.L. wrote the

manuscript. C.H., Y.-R.C., J.D.S., P.L.F., J.M.B. and X.L. supervised the study. CORRESPONDING AUTHOR Correspondence to Xiaoxia Li. ETHICS DECLARATIONS COMPETING INTERESTS The authors

declare no competing interests. ADDITIONAL INFORMATION PEER REVIEW INFORMATION L. A. Dempsey was the primary editor on this article and managed its editorial process and peer review in

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EXTENDED DATA EXTENDED DATA FIG. 1 EXTENDED INFORMATION RELATED TO FIG. 1. A. Characteristics of patients, related to Fig. 1a. B. Extracellular acidification rate (ECAR), related to Fig. 1d.

Data represent mean ± SEM. Data represent one of five independent experiments with similar results. C. Coomassie blue staining of mitochondrial proteins, related to Fig. 1e. D.

Mitochondrial proteins were extracted from WT and _Il1r1_ KO; WT and _Irak2_ KO newly differentiated primary adipocytes treated with or without IL-1β for indicated time points and analyzed

by SDS-PAGE, followed by western blot analysis with indicated antibodies. Data represent one of five independent experiments with similar results. E. Octanoyl-carnitine oxidation-rate in

isolated mitochondria from WT and _Il1r1_ KO primary adipocytes treated with or without IL-1β (n=4). Student’s t-test (two-tailed) was performed. Data represent mean ± SEM. Source data

EXTENDED DATA FIG. 2 IL-1 STIMULATION DID NOT LEAD TO MITOCHONDRIA DYSFUNCTION. Newly differentiated primary adipocytes were treated with IL-1β for indicated time points. Cytosolic mtDNA

(n=3) (A) and cytosolic Ca2+ (n=4) (B) levels were measured. C. Western blot analysis of cytoplasmic and mitochondrial proteins from IL-1β or TNFα + cycloheximide (CHX) treated newly

differentiated primary adipocytes. Data represent one of five independent experiments with similar results. D. Newly differentiated primary adipocytes were treated with IL-1β or

trifluoromethoxy carbonylcyanide phenylhydrazone (FCCP) for indicated time points. Cellular reactive oxygen species (ROS) were measured using fluorescent microscopy and microplate reader

(n=5). E. Western blot analysis of cytoplasmic and mitochondrial proteins from oligomycin + antimycin (OA) treated newly differentiated WT and _Irak2_ KO primary adipocytes (pretreated with

IL-1β or PBS). Data represent one of five independent experiments with similar results. Densitometric analysis of LC3 I/LC3 III were listed a bar graph (n=3). A, B, E: Student’s t-test

(two-tailed) was performed. Data represent mean ± SEM. D: One-way ANOVA was performed. Data represent mean ± SEM. Source data EXTENDED DATA FIG. 3 EXTENDED INFORMATION RELATED TO FIG. 2. A.

Total lysates from iWAT, BAT and liver from _Myd88_FF and _Myd88_AKO mice were subjected to western blot analysis with indicated antibodies. Data represent one of five independent

experiments with similar results. B. Transmission electron microscopy analysis of iWAT sections from HFD-fed mice. Scale bars, 1 μm. Morphometric analysis of cristae area versus mitochondria

area in 40 randomly selected mitochondria per group. C. Coomassie blue staining of mitochondrial proteins, related to Fig. 2i. D. Octanoyl-carnitine oxidation-rate in isolated mitochondria

from BAT of HFD-fed mice with indicated genotypes (n=5). E. Rectal temperatures were measured for HFD-fed mice with indicated genotypes (n=5). B, D, E: Student’s t-test (two-tailed) was

performed. Data represent mean ± SEM. Source data EXTENDED DATA FIG. 4 EXTENDED INFORMATION RELATED TO FIG. 3. A. Western blot analysis of immune-precipitated IRAK2 from IL-1β treated

mitochondria and subjected to indicated amount of Lambda phosphatase (Lambda PP) treatment for 10 min. B. Wild-type and HA-tagged IRAK2 were restored in _Irak2_ KO adipocytes. Western blot

analysis of cytoplasmic and mitochondrial proteins from IL-1β treated cells. C. Immunogold staining of HA-tagged IRAK2 which was overexpressed in _Irak2_ KO, _Myd88_ KO, _Il1r1_ KO and _Phb_

KD cells with or without IL-1 treatment for 24h. Scale bars, 50 nm. D. Immunogold staining of HA-tagged MyD88 in newly differentiated primary adipocytes from _Myd88_-HA reporter mice. Mito:

mitochondrion; Cyto: cytosol. Scale bar, 200 nm. E. Co-immunoprecipitation (IP) analysis of TIMM50 and TOM20 in mitochondria of in newly differentiated primary adipocytes from Irak2-HA

reporter mice treated with IL-1β for indicated time points and followed by western blot analysis. A, B, E: Data represent one of five independent experiments with similar results. Source

data EXTENDED DATA FIG. 5 EXTENDED INFORMATION RELATED TO FIG. 3. A. Coomassie blue staining of mitochondrial proteins, related to Fig. 3d. B. Expression of indicated mRNAs in WT and _Irak2_

KO newly differentiated primary adipocytes treated with or without IL-1β for indicated time points (n=3). C. Extracellular acidification rate (ECAR), related to Fig. 3f. Data represent mean

± SEM. Data represent one of five independent experiments with similar results. D. Octanoyl-carnitine oxidation-rate in isolated mitochondria from WT and _Irak2_ KO primary adipocytes

treated with or without IL-1β for 24h (n=5). E. Secondary helical wheel structure of IRAK2 peptide (amino acid: 39–52) which contains mitochondrial localization signal (MLS). The picture was

generated by http://lbqp.unb.br/NetWheels/. F. Coomassie blue staining of mitochondrial proteins, related to Fig. 3k. G. Octanoyl-carnitine oxidation-rate in isolated mitochondria from

Flag-tagged wild-type and IRAK2 mito-mutant restored _Irak2_ KO adipocytes treated with or without IL-1β for 24h (n=5). B, D, G: Student’s t-test (two-tailed) was performed. Data represent

mean ± SEM. Source data EXTENDED DATA FIG. 6 EXTENDED INFORMATION RELATED TO FIG. 4. A. Body weight of WT and _Irak2_ KO mice on HFD feeding (n=6 females per group). B. Glucose tolerance

test (GTT) and insulin tolerance test (ITT) were performed on HFD-fed of WT and _Irak2_ KO mice (n=5 females per group). C. H&E staining of iWAT sections from HFD-fed of WT and _Irak2_

KO mice. Cell size was quantified (3 views per slide, 3 sections per mouse, n=5). D. H&E staining of BAT sections from HFD-fed of WT and _Irak2_ KO mice. C, D: scale bars, 150 μm. A, B:

Two-way ANOVA, followed by post hoc analysis was performed. Data represent mean ± SEM. C: Student’s t-test (two-tailed) was performed. Data represent mean ± SEM. Source data EXTENDED DATA

FIG. 7 EXTENDED INFORMATION RELATED TO FIG. 5. A. Octanoyl-carnitine oxidation-rate in isolated mitochondria from BAT of HFD-fed mice with indicated genotypes (n=5). B. Various pictures of

immunogold staining of HA-tagged IRAK2 in BAT sections of HFD-fed _Irak2_-HA reporter mice, related to Fig. 5c Scale bars, 200nm. C. Coomassie blue staining of mitochondrial proteins,

related to Fig. 5e. D-E. Immunohistochemical staining of RFP in BAT sections of HFD-fed WT and _Irak2_ KO Ucp1-luc/tdTomato reporter mice. Luciferase enzymatic activity in lysates from BAT

(d) and iWAT (e) (n=8). F, G. HFD-fed WT and _Irak2_ KO Ucp1-luc/tdTomato reporter mice injected with CL-316,243 for 3 days. Immunohistochemical staining of RFP in BAT sections and

Luciferase enzymatic activity in lysates from BAT (L) and iWAT (M) (n=6). D, F: Scale bars, 150 nm. A, D–G: Student’s t-test (two-tailed) was performed. Data represent mean ± SEM. Source

data EXTENDED DATA FIG. 8 EXTENDED INFORMATION RELATED TO FIG. 6. A. Co-immunoprecipitation (IP) analysis of HA-tagged IRAK2, in mitochondria of in newly differentiated primary adipocytes

from _Irak2_-HA reporter mice treated with IL-1β for indicated time points and followed by western blot analysis. Data represent one of five independent experiments with similar results. B,

C: Densitometric analysis of western blots in Fig. 6a, b. B. Signals corresponding to IP OPA1 were used and normalized to Mito input OPA1 in Fig. 6a (n=3). C. Signals corresponding to IP

IRAK2 were used and normalized to Mito input IRAK2 in Fig. 6b (n=3). D, Mitochondrial proteins were extracted Ctrl and _Phb_ KD primary adipocytes treated with IL-1β for indicated time

points and analyzed by BN-PAGE, followed by Western blot analysis with anti-OxPhos cocktail antibodies. HMW SCs: high molecular weight super-complexes. E. [1-14C]-palmitic acid

oxidation-rate F. Octanoyl-carnitine oxidation-rate in isolated mitochondria from WT and _Phb_ KD primary adipocytes, treated with or without IL-1β for 24h (n=5). G. The activities of

respiratory complexes H. Octanoyl-carnitine oxidation-rate in the isolated mitochondria in mitochondria of non-targeting siRNA (WT and _Irak2_ KO) and PHB siRNA transfected (WT/_Phb_ KD,

_Irak2_ / _Phb_ KD) WT and _Irak2_ KO primary adipocytes treated with/without IL-1β (n=6). I. The activities of respiratory complexes J. Octanoyl-carnitine oxidation-rate in the isolated

mitochondria in mitochondria of empty vector (WT and _Irak2_ KO) and PHB cDNA transfected (WT/PHB and _Irak2_ KO/PHB) WT and _Irak2_ KO primary adipocytes treated with/without IL-1β (n=6).

B, C, E–J: Student’s t-test (two-tailed) was performed. Data represent mean ± SEM. Source data EXTENDED DATA FIG. 9 EXTENDED INFORMATION RELATED TO FIG. 6. A. Immunogold staining of

HA-tagged IRAK2 and kinase-inactive (KI) mutant which were overexpressed in _Irak2_ KO cells with or without IL-1β treatment for 24h. Scale bars, 50 nm. B. Coomassie blue staining of

mitochondrial proteins, related to Fig. 6d. C. Extracellular acidification rate (ECAR), related to Fig. 6g. Data represent mean ± SEM. Data represent one of five independent experiments with

similar results. D. Octanoyl-carnitine oxidation-rate in isolated mitochondria from WT and _Irak2_ KI primary adipocytes, treated with or without IL-1β for 24h (n=5). Student’s t-test

(two-tailed) was performed. Data represent mean ± SEM. Source data EXTENDED DATA FIG. 10 EXTENDED INFORMATION RELATED TO FIG. 7 & 8. A. Rectal temperatures were measured for HFD-fed mice

with indicated genotypes (n=5) B. Coomassie blue staining of mitochondrial proteins, related to Fig. 7g. C. Targeting vector design for generation of a novel mouse strain with exon 1 of

Irak2 flanked by _lox_P sites. D. Total lysates from iWAT, BAT and liver from _Irak2_FF and _Irak2__AKO_ mice were subjected to western blot analysis with indicated antibodies. Data

represent one of five independent experiments with similar results. E. Rectal temperatures were measured for HFD-fed mice with indicated genotypes (n=5). f. Coomassie blue staining of

mitochondrial proteins, related to Fig. 8g. A, C: Student’s t-test (two-tailed) was performed. Data represent mean ± SEM. Source data SUPPLEMENTARY INFORMATION REPORTING SUMMARY SOURCE DATA

SOURCE DATA FIG. 1 Unprocessed western blots for Fig. 1c,e. SOURCE DATA FIG. 1 Statistical raw data for Fig. 1a,b,d–g. SOURCE DATA FIG. 2 Unprocessed western blots for Fig. 2i. SOURCE DATA

FIG. 2 Statistical raw data for Fig. 2a–k. SOURCE DATA FIG. 3 Unprocessed western blots for Fig. 3a,b,d,i–k. SOURCE DATA FIG. 3 Statistical raw data for Fig. 3d–h,k–n. SOURCE DATA FIG. 4

Statistical raw data for Fig. 4a–i. SOURCE DATA FIG. 5 Unprocessed western blots for Fig. 5e. SOURCE DATA FIG. 5 Statistical raw data for Fig. 5b,d–i. SOURCE DATA FIG. 6 Unprocessed western

blots for Fig. 6a–d,i,j. SOURCE DATA FIG. 6 Statistical raw data for Fig. 6d–h. SOURCE DATA FIG. 7 Unprocessed western blots for Fig. 7g. SOURCE DATA FIG. 7 Statistical raw data for Fig.

7a–d,f–i. SOURCE DATA FIG. 8 Unprocessed western blots for Fig. 8g. SOURCE DATA FIG. 8 Statistical raw data for Fig. 8a–d,f–i. SOURCE DATA EXTENDED DATA FIG. 1 Unprocessed western blots and

gels for Extended Data Fig. 1c,d. SOURCE DATA EXTENDED DATA FIG. 1 Statistical raw data for Extended Data Fig. 1b,e. SOURCE DATA EXTENDED DATA FIG. 2 Unprocessed western blots for Extended

Data Fig. 2c,e. SOURCE DATA EXTENDED DATA FIG. 2 Statistical raw data for Extended Data Fig. 2a,b,d,e. SOURCE DATA EXTENDED DATA FIG. 3 Unprocessed western blots and gels for Extended Data

Fig. 3a,c. SOURCE DATA EXTENDED DATA FIG. 3 Statistical raw data for Extended Data Fig. 3b–e. SOURCE DATA EXTENDED DATA FIG. 4 Unprocessed western blots for Extended Data Fig. 4a,b,e. SOURCE

DATA EXTENDED DATA FIG. 5 Unprocessed gels for Extended Data Fig. 5a,f. SOURCE DATA EXTENDED DATA FIG. 5 Statistical raw data for Extended Data Fig. 5b–d,g. SOURCE DATA EXTENDED DATA FIG. 6

Statistical raw data for Extended Data Fig. 6a–c. SOURCE DATA EXTENDED DATA FIG. 7 Unprocessed gels for Extended Data Fig. 7c. SOURCE DATA EXTENDED DATA FIG. 7 Statistical raw data for

Extended Data Fig. 7a,d–g. SOURCE DATA EXTENDED DATA FIG. 8 Unprocessed western blots for Extended Data Fig. 8a,d. SOURCE DATA EXTENDED DATA FIG. 8 Statistical raw data for Extended Data

Fig. 8b,c,e–i. SOURCE DATA EXTENDED DATA FIG. 9 Unprocessed gels for Extended Data Fig. 9b. SOURCE DATA EXTENDED DATA FIG. 9 Statistical raw data for Extended Data Fig. 9c,d. SOURCE DATA

EXTENDED DATA FIG. 10 Unprocessed western blots and gels for Extended Data Fig. 10b,d,f. SOURCE DATA EXTENDED DATA FIG. 10 Statistical raw data for Extended Data Fig. 10a,e. RIGHTS AND

PERMISSIONS Reprints and permissions ABOUT THIS ARTICLE CITE THIS ARTICLE Zhou, H., Wang, H., Yu, M. _et al._ IL-1 induces mitochondrial translocation of IRAK2 to suppress oxidative

metabolism in adipocytes. _Nat Immunol_ 21, 1219–1231 (2020). https://doi.org/10.1038/s41590-020-0750-1 Download citation * Received: 06 June 2019 * Accepted: 25 June 2020 * Published: 10

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