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Nature Cardiovascular ResearchPhysiologyPeer-reviewed journal article

Targeting immunometabolic pathways with AZD1656 alleviates inflammation and metabolic dysfunction in type 2 diabetic cardiomyopathy

Stephanie Anderson, Anja KarlstaedtiD, Megan YoungiD, Loucia Karatzia, Fenn Cullen, Jianmin CheniD, Caroline E. O’RiordaniD, Michael R. BarnesiD, Zorana ŠtakaiD, Lauren J. Albee, Conor Garrod-KetchleyiD, Sanushi Dambure, Hiran A. PragiD, Filip Cvetko, Jack J. J. J. MilleriD, Christoph ThiemermanniD, Andrew J. M. Lewis, Michael P. MurphyiD, David M. SmithiD, Siân M. HensoniD, Damian J. TyleriD, Dunja AksentijevićiD

PDFDOIPreprintData

In-depth summary

Claude Sonnet 4.6 · xhigh effort · 8 Nov 2026 · from the full text of version 2 (7 Nov 2025) · CC BY 4.0

In db/db mice, six weeks of AZD1656 improved diastolic function, cardiac metabolism and infarct size, with increased Treg infiltration and less fibrosis, without glycaemic change.

Diabetic cardiomyopathy has no therapy designed to target cardiac metabolism, and anti-inflammatory agents have mostly failed in trials because they hit innate, acute inflammation. This work tests whether AZD1656, a glucokinase activator with reported Treg-modulating effects, links T-cell cardiac inflammation to metabolic remodelling in db/db mice.

Key results

  1. ShownSix weeks of AZD1656 improved diastolic function indices in db/db mice, reduced infarct size after ischaemia-reperfusion and improved post-ischaemic left ventricular developed pressure (LVDP) recovery in perfused hearts.

    E/A abnormal-category Fisher's exact test: control vs db/db P<0.0003, AZD vs db/db P<0.036, AZD vs control P<0.185 (control n=17, db/db n=17, AZD n=14). Infarct size n=5/group for db/db and AZD, control n=3. · Data analysis and statistics; AZD1656 treatment improves diastolic function in…; Figure legends

  2. ShownAZD1656 moved the db/db cardiac metabolomic profile close to control and increased in vivo pyruvate-to-bicarbonate conversion (a PDH flux readout), which was strongly reduced in untreated db/db hearts. CardioNet modelling predicted lower oxygen consumption.

    Hyperpolarized 13C MRS: db/db had 95% lower PDH flux than control; AZD1656 raised bicarbonate conversion (control n=12, db/db n=16, AZD n=10). 1H NMR PCA: AZD1656 hearts indistinguishable from control (n=6/group). CardioNet: 2-fold lower predicted O2 consumption. · AZD1656 treatment improves cardiac metabolism in…; Figure legends

  3. ShownAZD1656 shifted the db/db myocardial immune landscape: more Foxp3+ Tregs, fewer infiltrating T cells and B cells, and less interstitial fibrosis. Dendritic cells, neutrophils and macrophages were unchanged.

    Untreated db/db hearts had no Treg increase; AZD1656 raised myocardial CD4+CD25+FoxP3+ frequency (n=9/group). Masson trichrome fibrosis n=5 control, 4 db/db, 4 AZD. Splenic CD3+ abundance was unchanged. No effect sizes are given in the text. · AZD1656 treatment reduces T-cell mediated…; Figure legends

  4. ShownThe cardiac benefits were not accompanied by improvements in systemic metabolism: body weight, glucose, insulin, liver readouts and whole-body fat mass did not improve, while circulating free fatty acids fell significantly.

    Weight showed no improvement with AZD1656 (23 control, 17 db/db, 20 treated mice); fasting glucose was unchanged (9 per group); free fatty acids fell significantly (11 control, 9 db/db, 5 treated); lean mass rose only slightly (5 per group). · AZD1656 treatment effects are not mediated by…; Figure legends

  5. ShownIn db/db mice, 19 of the 30 most upregulated cardiac genes were pro-inflammatory, and AZD1656 reversed much of this signature, including Nlrp3 expression and the leukocyte extravasation, HIF1α and NRF2 pathways.

    DEGs (FC>1.5, P<0.05): 1379 db/db vs control, 503 AZD vs db/db, 1480 control vs AZD; 19 of the top 30 upregulated db/db genes pro-inflammatory vs 8 for AZD vs db/db; Nlrp3 2.5-fold up in db/db (n=6 hearts/group). · Treatment with AZD1656 normalizes the expression…; Figure legends

Methods, data and reproducibility

Male db/db mice received AZD1656 in diet (30 mg/kg/day) from 14 to 20 weeks of age and were compared with untreated db/db and lean controls using MRI, echocardiography, Langendorff ischaemia-reperfusion, hyperpolarized 13C MRS, 1H NMR, lipidomics, CardioNet flux modelling, flow cytometry, histology and cardiac MACE-seq.

  • Male lean controls, untreated db/db and AZD1656-treated db/db mice (study totals across cohorts; per-assay n varied) · mouse · n = Control 64, db/db 63, AZD1656 57
  • Cardiac 3' RNA-seq (MACE-seq) of heart tissue from the three groups · mouse · n = 6 hearts/group · E-MTAB-13849
  • Cardiac 1H NMR metabolomics feeding CardioNet flux balance analysis · mouse · n = 6/group
  • Cardiac non-targeted lipidomics by LC-MS/MS · mouse · n = 5/group
  • In vivo hyperpolarized 13C pyruvate MRS cardiac metabolic flux · mouse · n = Control 12, db/db 16, AZD 10
  • Echocardiography diastolic assessment (E/A analysis) · mouse · n = Control 17, db/db 17, AZD 14
  • Plasma XXL cytokine array · mouse · n = Control 10, db/db 12, AZD 10
  • High-fat-diet-fed pre-diabetic C57BL/6 male mice used for temporal sequence of metabolic vs immune changes · mouse · n = 18
Methods & tools
AZD1656 (glucokinase activator), CardioNet, GUROBI solver, MACE-Seq, g:Profiler, Ingenuity Pathway Analysis, Metascape, PLS-DA (mdatools), Hyperpolarized 13C MRS, Flow cytometry (FACSAria III, FlowJo), Langendorff perfusion with TTC staining, Fisher's exact test
Reagents & environment
Python with Matplotlib 3.4.3, Pandas 1.3.4, NumPy 1.20.3; GUROBI 9.1.2; RStudio 2022.12.0; GraphPad Prism v9; g:Profiler e108_eg55_p17_0254fbf; Cutadapt 3.4; FlowJo v10; VevoLab 5.5.1
Reproducibility gaps
No analysis code is provided. The Dryad link is a placeholder-style private share link. Some data are only in supplementary files or available from the corresponding author on request.
Limitations (6)
  • Only heart-resident CD4+ T cells were studied; peripheral blood and lymphoid tissues were not assessed and the systemic TCR repertoire was not sequenced. · stated by the authors
  • The causal role of CD4+ T cells in dbCM remains untested; CD4+ T-cell-deficient or antibody-depletion experiments are proposed for future work. · stated by the authors
  • AZD1656 reduced circulating free fatty acids, which may have contributed to the improved myocardial metabolism and inflammation, so systemic lipid effects are not excluded. · stated by the authors
  • Parameters were measured over a 6-year study in multiple cohorts, so sample sizes varied between assays. · stated by the authors
  • Several key endpoints rest on small groups (infarct size n=5/group with control n=3; fibrosis n=4 in db/db and AZD) in male mice of a single genetic model, limiting power and generality. · noted in this summary
  • The Treg-mediated mechanism is inferred from changes in cell frequency, cytokines and transcripts; no Treg depletion or transfer experiment is reported, so other drug targets (hepatic or beta-cell glucokinase) are not excluded. · noted in this summary

Read this if you study cardiac immunometabolism or diabetic cardiomyopathy and want preclinical evidence linking a glucokinase activator to Treg infiltration, metabolic flux and infarct protection.

Citations
4
FWCI
19.99
Field percentile
top 1%

Abstract

Type 2 diabetes (T2D) precipitates diabetic cardiomyopathy (dbCM), a condition characterized by chronic inflammation, metabolic dysregulation and impaired cardiac performance. Here we show that the glucokinase activator AZD1656, originally developed for glycaemic control but later identified to have immunomodulatory effects, reverses cardiac dysfunction and metabolic remodelling in dbCM. In obese, hyperglycaemic db/db mice with diastolic dysfunction, six weeks of AZD1656 treatment improved myocardial performance, reduced infarct size and enhanced post-ischaemic recovery. Integrated metabolic, functional and histological analyses revealed restoration of mitochondrial metabolism and attenuation of fibrosis. Mechanistically, AZD1656 remodelled the cardiac immune landscape by promoting regulatory T-cell infiltration. These findings demonstrate a link between cardiac inflammation and metabolic remodelling in dbCM and highlight that modulation of immune cells and metabolism can protect the diabetic heart. Targeting immunometabolic pathways may therefore offer a therapeutic strategy to alleviate cardiac dysfunction and reduce infarct vulnerability in T2D

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