T cell-derived TNF drives myocarditis but is dispensable for antitumor efficacy of checkpoint immunotherapy
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ABSTRACT:
ORGANISM(S): Mus musculus
PROVIDER: GSE302512 | GEO | 2026/02/03
REPOSITORIES: GEO
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Project description:Objective: Immune checkpoint inhibitors (ICIs) have revolutionized cancer therapy but are frequently associated with immune-related adverse events, particularly in combination regimens. ICI-associated myocarditis (ICI-M) is rare but highly lethal, with a mortality rate approaching 50% and a notable risk of fatal ventricular arrhythmias. Prevention and effective management of ICI-M remain significant clinical challenges. This study aims to elucidate the immune landscape and spatial organization of immune cells in ICI-M using single-cell multi-omics in a preclinical mouse model. Methods: We utilized Ctla4+/–Pdcd1–/– mice, modeling human ICI-induced myocarditis. Single-cell RNA (scRNAseq) and T cell receptor (TCR) sequencing were performed on heart-infiltrating immune cells and peripheral blood mononuclear cells (PBMCs). Spatial transcriptomics was applied to map immune infiltration in cardiac tissue. Results: Approximately half of Ctla4+/–Pdcd1–/– mice died after 100 days, while Ctla4+/+ mice showed normal survival. ECG monitoring identified ventricular arrhythmias in 15% of Ctla4+/–Pdcd1–/– mice. Histology revealed severe myocardial inflammation and fibrosis. Cardiac scRNAseq data indicated that T cells were the predominant immune population in ICI-M hearts. TCR sequencing demonstrated substantial overlap in TCR clonotypes between cardiac-infiltrating and circulating CD8+ T cells. These TCR-shared CD8+ T cells exhibited an activated phenotype and were enriched in pathways associated with allograft rejection and inflammation. Spatial mapping revealed localization near the AV node and subendocardium, implicating potential CD8+ T cell–mediated conduction disruption. Conclusion: Clonal, activated CD8+ T cells contribute to arrhythmogenic myocarditis in ICI-M. Targeting specific TCR-peptide-MHC interactions may enable myocarditis prevention without impairing antitumor efficacy.
2025-06-11 | GSE299032 | GEO
Project description:Objective: Immune checkpoint inhibitors (ICIs) have revolutionized cancer therapy but are frequently associated with immune-related adverse events, particularly in combination regimens. ICI-associated myocarditis (ICI-M) is rare but highly lethal, with a mortality rate approaching 50% and a notable risk of fatal ventricular arrhythmias. Prevention and effective management of ICI-M remain significant clinical challenges. This study aims to elucidate the immune landscape and spatial organization of immune cells in ICI-M using single-cell multi-omics in a preclinical mouse model. Methods: We utilized Ctla4+/–Pdcd1–/– mice, modeling human ICI-induced myocarditis. Single-cell RNA (scRNAseq) and T cell receptor (TCR) sequencing were performed on heart-infiltrating immune cells and peripheral blood mononuclear cells (PBMCs). Spatial transcriptomics was applied to map immune infiltration in cardiac tissue. Results: Approximately half of Ctla4+/–Pdcd1–/– mice died after 100 days, while Ctla4+/+ mice showed normal survival. ECG monitoring identified ventricular arrhythmias in 15% of Ctla4+/–Pdcd1–/– mice. Histology revealed severe myocardial inflammation and fibrosis. Cardiac scRNAseq data indicated that T cells were the predominant immune population in ICI-M hearts. TCR sequencing demonstrated substantial overlap in TCR clonotypes between cardiac-infiltrating and circulating CD8+ T cells. These TCR-shared CD8+ T cells exhibited an activated phenotype and were enriched in pathways associated with allograft rejection and inflammation. Spatial mapping revealed localization near the AV node and subendocardium, implicating potential CD8+ T cell–mediated conduction disruption. Conclusion: Clonal, activated CD8+ T cells contribute to arrhythmogenic myocarditis in ICI-M. Targeting specific TCR-peptide-MHC interactions may enable myocarditis prevention without impairing antitumor efficacy.
2025-06-11 | GSE298932 | GEO
Project description:Immune checkpoint inhibitors (ICIs) are effective against many cancers but can also cause immune-related adverse events. Although rare, ICI-mediated myocarditis has a high fatality rate of up to 40% and is also associated with heart failure and life-threatening arrhythmias. We characterized a population of CXCL9+CXCL10+ macrophages and CXCR3hi CD8+ T-cells in the hearts of mice with myocarditis and elucidated chemokine crosstalk between the CXCR3hi CD8+ effector T-cells and the CXCL9+CXCL10+ macrophages in the heart. Depletion of macrophages or blockade of CXCR3 both resulted in significantly decreased immune cell infiltration in the hearts of mice. Similarly, CXCR3 blockade decreased immune cell infiltration into the heart,, thus posing CXCR3 and its ligands as an attractive therapeutic target. Additionally, an in vitro transwell assay showed that selective blockade of CXCR3 and its ligand CXCL10 significantly decreased CD8+ T-cell migration towards macrophages, implicating this interaction in T-cell cardiotropism towards cardiac macrophages. These findings were compared with cardiac biopsies from patients with ICI myocarditis that also demonstrated infiltrating CXCR3+ lymphocytes and CXCL9+/CXCL10+ macrophages. In both mouse cardiac immune cells and patient peripheral blood immune cells, T-cell receptor (TCR)-sequencing revealed expanded TCRs that correlated with CXCR3hi CD8+ T-cells. The identification of a CXCR3-specific T-cell and macrophage interaction as important in the pathogenesis of ICI myocarditis offers a new potential target for a chemokine/chemokine receptor inhibition-focused form of therapy in this disease.
2025-01-31 | GSE250056 | GEO
Project description:Immune checkpoint inhibitors (ICIs), antibodies targeting PD-1/PD-L1 or CTLA4 have revolutionized cancer management but are associated with devastating immune-related adverse events (irAEs) including myocarditis. The main risk factor for ICI myocarditis is the use of combination PD-1 and CTLA4 inhibition. ICI-myocarditis is often fulminant and is pathophysiologically characterized by myocardial infiltration of T lymphocytes and macrophages. While much has been learned regarding the role of T-cells in ICI-myocarditis, little is understood regarding the identity, transcriptional diversity, and functions of infiltrating macrophages. We employed an established murine ICI myocarditis model (Ctla4+/-Pdcd1-/- mice) to explore the cardiac immune landscape using single-cell RNA-sequencing, immunostaining, and molecular imaging. We observed marked increases in CCR2+ monocyte-derived macrophages and CD8+ T-cells in this model. The macrophage compartment was heterogeneous and displayed marked enrichment in an inflammatory CCR2+ subpopulation expressing Cxcl9, Cxcl10, Gbp2b, and Fcgr4 that originated from CCR2+ monocytes. Importantly, a similar macrophage population expressing CXCL9, CXCL10, and CD16α (human homologue of mouse FcgR4) were found selectively in patients with ICI myocarditis compared to other forms of heart failure and myocarditis. In silico prediction of cell-cell communication suggested interactions between T-cells and Cxcl9+Cxcl10+ macrophages via IFN-γ and CXCR3 signaling pathways. Depleting CD8+T-cells and blockade of IFN-γ signaling blunted the emergence of Cxcl9+Cxcl10+ macrophages in the heart and attenuated myocarditis suggesting that this interaction was necessary for disease pathogenesis. Collectively, these data demonstrate that ICI-myocarditis is associated with the emergence of a specific population of inflammatory macrophages and suggest the possibility that IFN-γ blockade may serve as an effective treatment for this devastating condition.
2023-07-01 | GSE227437 | GEO
Project description:Immune checkpoint inhibitors (ICIs) are monoclonal antibodies used widely to activate the immune system against tumor cells. Despite their therapeutic benefits, ICIs are known to cause immune-related adverse events (irAE) such as myocarditis, a rare but serious side effect with up to 50% mortality. To identify pathogenic immune subset(s) responsible for ICI myocarditis and other irAE, we performed single cell mass cytometry (CyTOF) on peripheral blood mononuclear cells from 40 patients and controls with irAE including four patients with ICI myocarditis. We also profiled 15 patients/controls using single cell RNA sequencing (scRNA-seq) with feature barcoding for surface marker expression confirmation. In both CyTOF/scRNAseq analyses, we found expansions of cytotoxic CD8+ T effector cells re-expressing CD45RA (Temra CD8+ cells) in ICI myocarditis patients compared to controls. Using T cell receptor sequencing, we demonstrated a significant clonal expansion of CD8+ Temra cells in myocarditis patients. Transcriptomic profiling of these Temra CD8+ clones confirmed their highly activated and cytotoxic phenotype with expression of granzyme/perforin. Longitudinal data revealed the progression of these Temra CD8+ cells into an exhausted phenotype two months after diagnosis of myocarditis and after treatment with glucocorticoids. Differential expression of several proinflammatory chemokines (CCL4/CCL4L2/CCL5) was uncovered in the clonally expanded Temra CD8+ cells and ligand-receptor analysis implicated their regulation of other inflammatory cells such as monocytes and NK cells. These data suggest that the therapeutic modulation of these chemokines may serve as an attractive strategy for reducing life-threatening irAE in ICI-treated cancer patients.
2022-06-29 | GSE180045 | GEO
Project description:Immune checkpoint inhibitors (ICIs) have been increasingly used in combination for cancer treatment, but are particularly associated with myocarditis. Here, we report that tumor-bearing mice exhibited response to treatment with combinatorial anti-PD-1 (programmed cell death 1) and anti-CTLA-4 (Cytotoxic T-lymphocyte Antigen-4) antibodies but also presented with cardiovascular toxicities observed clinically with ICI therapy, including myocarditis and arrhythmia. Female mice were preferentially affected with myocarditis compared to male mice, consistent with a previously described genetic model of ICI-myocarditis as well as emerging clinical data. Mechanistically, myocardial tissue from ICI-treated mice, the genetic mouse model, and human heart tissue from affected patients with ICI-myocarditis all exhibited downregulation of MANF (Mesencephalic Astrocyte Derived Neurotrophic Factor) and HSPA5 (Heat Shock 70kDa Protein 5) in the heart; this downregulation was particularly striking in female mice. ICI-myocarditis was amplified by heart-specific genetic deletion of mouse Manf and was attenuated by administration of recombinant MANF protein, suggesting a causal role. Ironically, both MANF and HSPA5 were transcriptionally induced by liganded estrogen receptor-βbeta and inhibited by androgen receptor. However, ICI treatment reduced serum estradiol concentration to a greater extent in female compared to male mice. Treatment with an estrogen receptor β-specific agonist as well as androgen depletion therapy attenuated ICI-associated cardiac effects. Taken together, our data suggest that ICI treatment inhibits the estradiol-dependent expression of MANF/HSPA5 in the heart, curtailing the cardiomyocyte response to immune injury. This endocrine-cardiac-immune pathway offers new insights into the mechanisms of sex differences in cardiac disease and may offer treatment strategies for ICI-myocarditis.
2022-09-18 | GSE213468 | GEO
Project description:PURPOSE: The goal of this study was to determine the gene expression networks regulated by tumor necrosis factor receptor 2 (TNFR2, or Tnfrsf1b) and to evaluate their potential bearing on immune cell subsets and inflammatory bowel disease (IBD). METHODS: mRNA-seq was performed on isolated distal colons from TNFR2-knockout and wildtype mice. Differentially expressed transcripts were compared to human ulcerative colitis microarray datasets on Gene Expression Omnibus and to mouse immunological expression datasets at the Immunological Genome Project. RESULTS: We identified 252 mouse transcripts whose expressions were significantly altered by the loss of TNFR2. The majority of these transcripts (228 of 252, ~90%) were downregulated in TNFR2-/- colons. TNFR2-regulated genes were able to positively discriminate between ulcerative colitis patients and healthy individuals with ~80% accuracy. Many TNFR2-regulated genes were also highly expressed in CD8+ T cells. CONCLUSIONS: Downregulation of TNFR2 is associated with a gene expression profile that is prominent in IBD and supportive of the role of CD8+ T cells in IBD pathogenesis. MANUSCRIPT ABSTRACT: Increased tumor necrosis factor (TNF) production has been associated with inflammatory bowel disease (IBD), and anti-TNF therapy is a common therapeutic for this patient population. However, the role of TNF or its receptors (TNFR1 and TNFR2) in the immunopathogenesis of inflammatory bowel disease (IBD) remains unclear. Here we report that TNFR2 is protective in spontaneous (IL-10 knockout) and chemically (azoxymethane/dextran sodium sulfate)-induced mouse models of colitis and colitis-associated cancer. Mechanistically, TNFR2-deficiency in hematopoietic cells significantly increased incidence and severity of colitis and colitis-associated cancer characterized by a selective expansion of CD8+ T cells. We identified TNFR2-regulated genes in the colon that were specific for CD8+ T cells, interacted with multiple IBD risk genes, and are important regulators of CD8+ T cell biology. TNFR2 regulated CD8+ T-cell-specific genes that act as genetic susceptibility modifiers for IBD to mitigate the development of a pro-colitogenic milieu. Antibody-mediated depletion of CD8+ T cells prevented colonic inflammation and significantly reduced pathology in IL10-/-/TNFR2-/- deficient mice. Furthermore, adoptive transfer of TNFR2-/- naïve CD8+ T cells resulted in more severe disease than with wildtype naïve CD8+ T cells. Our findings provide insight into the disease modifier role of TNFR2 in the immunopathogenesis of IBD through the modulation of CD8+ T cell responses and support future investigation of this therapeutic target, especially in the subset of IBD patients with CD8+ T-cell dysfunction. Total RNA from distal colons of 8 week-old male wildtype C57Bl/6 and TNFR2-/- mice (n=3 each) was isolated using the PureLink RNA kit (Ambion, Life Technologies). RNA samples were submitted to the Genomic Services Lab at the HudsonAlpha Institute for Biotechnology (Huntsville, AL) for multiplex library preparation, mRNA enrichment, and sequencing. Sequencing was performed to an average depth of 50M paired-end 50bp reads per sample (HiSeq, Illumina, San Diego, CA). Data files containing raw reads were aligned to the mouse genome using Tophat2/Bowtie2. Alignments were assembled into transcript representations with Cufflinks, and statistical tests for differential expression were performed with Cuffdiff 2. An adjusted P value < 0.05 (q<0.05) from the Cuffdiff 2 output was used as the cutoff for statistical significance.
2015-08-11 | E-GEOD-65408 | biostudies-arrayexpress
Project description:The heterogenicity of hepatocellular carcinoma (HCC) remains a key obstacle in turning the majority of ‘immune-cold’ tumors ‘hot’ for effective immune checkpoint inhibitors (ICIs). Through analyzing the naturally-existed ‘hot’ HCC variants, we identified fas-associated death domain (FADD) as a key molecule upregulated in patients with dense tumor-filtrating CD8+T cells and better response to ICIs. Apart from the canonical role in apoptosis pathway, our data showed that CRISPR knockout of hepatoma-intrinsic Fadd led to increased tumor weights in immunocompetent but not immunodeficient mice, accompanied with decreased numbers and IFN-γ/TNF-ɑ production of tumor-filtrating CD8+T cells. Mechanistically, phosphorylated FADD translocated into cell nucleus, where it interacted with Sam68 to upregulate NF-κB transcription of CCL5, thereby promoted CD8+T cell tumor infiltration. Interestingly, anti-PD1 triggered FADD phosphorylation by CD8+T cell-derived IFN-γ/TNF-ɑ in ICI-sensitive, but not resistant tumors. Sequential FADD activation through genetic or pharmacologic approaches to orchestrate p-FADD-CD8+T cell axis therefore overcame ICI resistance in ICI-resistant orthotopic and spontaneous HCC mouse models in vivo. Taken together, our findings may provide insights into the combinatory immunotherapy approaches for the majority of HCC patients in the future.
2025-07-07 | PXD057519 | Pride
Project description:Myocarditis has emerged as an immune-related adverse event of immune-checkpoint inhibitor (ICI) cancer therapy associated with significant mortality. Our new NOD-cMHCI/II-/-.DQ8 mouse strain expresses human HLA-DQ8 in the absence of classical murine MHC class I and II. These mice are highly susceptible to myocarditis and acute heart failure following anti-PD-1 ICI-induced treatment. Additionally, anti-PD-1 administration accelerates skeletal muscle myositis development. Using RNA sequencing analyses we performed a thorough characterization of cardiac and skeletal muscle infiltrating T-cells, identifying shared and unique characteristics of both populations.
2024-02-02 | GSE248625 | GEO
Project description:Immune checkpoint inhibitor-induced myocarditis (ICI-m) is a highly fatal cancer treatment complication with challenging diagnosis and invasive diagnostic methods. We developed a plasma cell-free mRNA (cf-mRNA) platform for noninvasive characterization of the immune and myocardial landscape in ICI-m. We sequenced bulk cell-free total RNA from plasma of cancer patients treated with ICIs: without immune-related adverse events (irAE) (group A, n=5), with noncardiac irAE (group B, n=7), and with ICI-m with/without noncardiac irAE (group C, n=10). Publicly available healthy control cfRNA (n=30) served as external controls. Cell type-specific gene panels from single cell RNA-sequencing of circulating immune and cardiac cells were used for cellular deconvolution. Cancer and inflammatory pathways were enriched in ICI-treated patients versus controls. Cardiac pathways and conduction system genes were significantly enriched in group C versus A and B. Temra CD8+ T cell and cardiomyocyte signature scores, with CCL5 and MYH6 expression, were elevated in group C. Our custom ICI-m classifier, containing three cardiac- and three immune cell-specific genes, effectively differentiated group C from A (AUC 0.903, 95% CI 0.858-0.903) and B patients (AUC 0.983, 95% CI 0.970-0.983), outperforming unsupervised feature selection. outperforming unsupervised feature selection. This platform captures disease-specific signatures enabling precision diagnostics for inflammatory diseases.
2025-05-24 | GSE296680 | GEO