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JACC Basic to Translational ScienceImmunologyPeer-reviewed journal article

Interleukin-1β Drives Disease Progression in Arrhythmogenic Cardiomyopathy

Vinay R. PennaiD, Junedh AmruteiD, Morgan EngeliD, Emily A. ShieliD, Waleed Farra, Elisa Nicole CannoniD, Colleen Leu-TurneriD, Pan MaiD, Ana VillanuevaiD, Hae-Won ShiniD, Alekhya ParvathaneniiD, Joanna JageriD, Carlos Bueno‐BetíiD, Angeliki AsimakiiD, Kory J. LavineiD, Jeffrey E. SaffitziD, Stephen P. ChelkoiD

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Abstract

Arrhythmogenic cardiomyopathy (ACM) is a genetic form of heart failure that affects 1 in 5000 people globally and is caused by mutations in cardiac desmosomal proteins including PKP2, DSP, and DSG2. Individuals with ACM suffer from ventricular arrhythmias, sudden cardiac death, and heart failure. There are few effective treatments and heart transplantation remains the best option for many affected individuals. Here we performed single nucleus RNA sequencing (snRNAseq) and spatial transcriptomics on myocardial samples from patients with ACM and control donors. We identified disease-associated spatial niches characterized by co-existence of fibrotic and inflammatory cell types and failing cardiac myocytes. The inflammatory-fibrotic niche co-localized to areas of cardiac myocyte loss and was comprised of FAP (fibroblast activation protein) and POSTN (periostin) expressing fibroblasts and macrophages expressing NLRP3 (NLR family pyrin domain containing 3) and NFB activated genes. Using homozygous Desmoglein-2 mutant (Dsg2mut/mut) mice, we identified analogous populations of Postn expressing fibroblasts and inflammatory macrophage populations that co-localized within diseased areas. Detailed single cell RNA sequencing analysis of inflammatory macrophage subsets that were increased in ACM samples revealed high levels of interleukin-1{beta} (Il1b) expression. To delineate the possible benefit of targeting IL-1{beta} in ACM, we treated Dsg2mut/mut mice with an anti-IL-1{beta} neutralizing antibody and observed attenuated fibrosis, reduced levels of inflammatory cytokines and chemokines, preserved cardiac function, and diminished conduction slowing and automaticity, key mechanisms of arrhythmogenesis. These results suggest that currently approved therapeutics that target IL-1{beta} or IL-1 signaling may improve outcomes for patients with ACM.

In-depth summary

Claude Opus 5.5 · xhigh effort · 8 Nov 2026 · from the full text of version 1 (17 Dec 2024) · CC BY 4.0

Human arrhythmogenic cardiomyopathy hearts harbor a lesion niche of POSTN fibroblasts and NLRP3+ inflammatory macrophages; IL-1β neutralization in Dsg2-mutant mice reduced fibrosis, dysfunction, and ventricular ectopy.

Arrhythmogenic cardiomyopathy (ACM), driven mostly by desmosomal variants such as PKP2, DSP and DSG2, has few effective therapies beyond transplantation, and the cellular makeup of its inflammatory lesions and their key mediators were largely undefined. Extending earlier work implicating NFκB and CCR2+ monocyte recruitment in Dsg2 mutant mice, this study maps human lesion niches and tests IL-1β, a target of approved drugs, as a causal driver.

Key results

  1. ShownWeekly anti-IL-1β neutralizing antibody given to Dsg2 mutant mice from 8 weeks of age, before overt remodeling, improved left ventricular ejection fraction, reduced premature ventricular contractions and fibrotic area, and lowered inflammatory and profibrotic cytokines relative to isotype control.

    Dosing 1mg/kg/week antibody vs 10mg/kg/week isotype for 8 weeks; significant LVEF gain and smaller Masson's trichrome fibrotic area, plus fewer PVCs and less ectopy (n=6, 10, and 9 per group; Brown-Forsythe and Welch ANOVA, P < 0.05 threshold); cytokine array showed lower CD14, CXCL2, CXCL9, IFNγ, osteopontin and POSTN. No effect sizes printed in text. · Targeting IL-1β attenuates disease…; IL1β treatment; Statistical analysis; Figure legends

  2. ShownHuman ACM left ventricles show expanded fibroblast, myeloid and T-cell compartments; POSTN/THBS4 fibroblasts (Fib3) and NLRP3+ macrophages (Mac4) are enriched and co-localize in a cardiomyocyte-depleted inflammatory-fibrotic spatial niche overlying histological lesions.

    snRNA-seq of 6 ACM (3 DSP, 3 PKP2) vs 12 donors: 14 cell types, 7 fibroblast and 6 myeloid states; Visium of 3 ACM vs 2 donors: 7 niches, Niche 4 concentrated in lesions with PROGENy NFκB and JAK-STAT enrichment and Tangram macrophage-fibroblast correlation; CD68+CCR2+ and FAP+ cells co-localized by immunofluorescence in two samples. · Single nucleus RNA (snRNAseq) sequencing reveals…; Spatial transcriptomics reveals formation of an…; snRNAseq reveals expansion of POSTN expressing…; Figure legends

  3. SuggestedDsg2 mutant mouse hearts contain transcriptionally analogous lesion populations: a Postn+ fibroblast cluster and inflammatory monocyte/macrophage states that score highly for orthologous human ACM signatures and express Il1b.

    Sorted-cell scRNA-seq of 6-week-old mutant vs WT hearts (n=3 per group): mFib3 (Postn, Comp) and mFib7 enriched in mutants; mMono1, mMac1 and mMac3 overrepresented; human myeloid ortholog signature highest in mMono1 and mMac3, which also expressed Il1b. Human fibroblast signature is reported in mFib2. · snRNAseq of Dsg2mut/mut mice reveals expansion…; Mouse 10x Single Cell preparation; Figure legends

  4. ShownIL-1β blockade partially reversed the ACM transcriptional program in cardiomyocytes, fibroblasts and endothelial cells and prevented cardiomyocyte RelA/p65 nuclear translocation, CCR2/CD68+ macrophage infiltration, lesional NLRP3/CASP1 staining and loss of junctional JUP and Cx43.

    iCell8cx SMART-seq Pro snRNA-seq of 16-week-old hearts (a platform offering >5,000 genes per nucleus): DE genes in all six cell types without composition change; many of the top 25 mutant-vs-WT genes fell after treatment; nuclear RelA and CCR2+ cells/mm^2 not elevated after treatment, and RelA-CCR2 correlation seen only in isotype mutants; n≥5 hearts/cohort/stain. · Treatment of Dsg2mut/mut mice with an anti-IL-1β…; Cardiac myocyte NFĸB nuclear localization and…; Targeting IL-1β attenuates disease…; Figure legends

  5. SuggestedDelaying treatment to 16 weeks, when dysfunction and biventricular fibrosis are established, still gave modest functional gains and a marked fibrosis reduction while isotype-treated mutants worsened, implying the largest benefit in the early inflammatory phase.

    Late-intervention cohort treated from 16 to 24 weeks with the same dosing (fig S6); described only qualitatively in the main text, without effect sizes, statistics or group sizes. · Targeting IL-1β attenuates disease…; Discussion; IL1β treatment

Methods, data and reproducibility

Paired 10x 5' v2 single-nucleus RNA-seq and FFPE Visium spatial transcriptomics of explanted ACM and donor left ventricles (Tangram deconvolution, niche clustering, PROGENy), cross-referenced to sorted-cell scRNA-seq of Dsg2 mutant mice, then anti-IL-1β antibody treatment assessed by echocardiography, ECG, histology, cytokine arrays and iCell8cx snRNA-seq.

  • Explanted left ventricular myocardium profiled by 10x snRNA-seq; ACM carriers of DSP or PKP2 variants and donor controls without heart disease · human · n = 6 ACM (3 DSP, 3 PKP2); 12 donor controls
  • FFPE left ventricular sections profiled by 10x Visium spatial transcriptomics (samples with DV200 > 40 selected) · human · n = 3 ACM (2 PKP2, 1 DSP); 2 donor controls
  • ACM myocardium immunostained for CD68, CCR2 and FAP to validate niche composition · human · n = two independent samples
  • FACS-sorted cardiac fibroblasts and myeloid cells from 6-week-old Dsg2 mut/mut and WT C57BL/6 hearts, 10x 3' v3.1 scRNA-seq · mouse · n = 3 per group
  • Early intervention: WT and Dsg2 mut/mut mice given anti-IL-1β or isotype from 8 weeks of age for 8 weeks; echo, ECG, histology, cytokine array · mouse · n = 6, 10, and 9 per group (Fig. 5)
  • Late intervention: WT and Dsg2 mut/mut mice treated with anti-IL-1β or isotype from 16 to 24 weeks of age · mouse
  • Nuclei from frozen early-intervention hearts at 16 weeks profiled by iCell8cx SMART-seq Pro snRNA-seq · mouse
  • Mouse heart sections immunostained for RelA, CCR2/CD68, JUP, Cx43, NLRP3 and CASP1 · mouse · n = n≥5 hearts/cohort/stain
Methods & tools
10x Genomics Chromium 5' v2 snRNA-seq and Visium FFPE, Cell Ranger v6.1 and Space Ranger, Seurat v4.0 with SCTransform, Scrublet, Tangram, stlearn, PROGENy, EnrichR, iCell8cx SMART-seq Pro with Cogent AP (Takara), Anti-IL-1β neutralizing antibody and isotype control (Novartis), Vevo F2 echocardiography and iWorx ECG analysis, GraphPad Prism v10; ImageJ 1.53e
Reagents & environment
Cell Ranger v6.1 (human GRCh38); Space Ranger; Seurat v4.0 in R with SCTransform; Scrublet; Tangram; stlearn; Cogent AP for iCell8cx data; GraphPad Prism v10; ImageJ version 1.53e.
Reproducibility gaps
Sequencing data announced for future GEO deposition with no accession given; analysis code available only on request; mouse scRNA-seq alignment described as 'mouse GRCh38', leaving the reference genome ambiguous.
Limitations (6)
  • Human tissue came from transplant explants at advanced disease, so the atlas cannot separate concealed from active ('hot') phases, and heterogeneous disease course and age at diagnosis may shape the cell states detected. · stated by the authors
  • Only PKP2 and DSP carriers were sequenced; variants in other ACM-linked genes, such as structural or Z-disc genes, may produce distinct cellular features. · stated by the authors
  • All human samples were left ventricular, so right ventricular adipocyte dysplasia, a major clinical feature of ACM, was not examined. · stated by the authors
  • The focus on IL-1β leaves other inflammatory pathways that likely contribute to ACM untested. · stated by the authors
  • Cohorts are small (spatial: 3 ACM vs 2 donors; mouse scRNA-seq: 3 per group), human composition shifts are reported without statistical testing, and the main text gives no numeric effect sizes for ejection fraction, PVC burden or fibrosis. · noted in this summary
  • Reporting inconsistencies weaken cross-species claims: the human fibroblast signature is attributed to mFib2 although the Postn cluster is mFib3, 12 myeloid states are stated but fewer listed, and isotype (10mg/kg/week) and anti-IL-1β (1mg/kg/week) doses differ. · noted in this summary

Read this if you study inherited cardiomyopathy, cardiac macrophage-fibroblast crosstalk, or repurposing IL-1 blockade, and want paired human single-nucleus/spatial atlases with mouse intervention data.

Citations
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top 1%

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