A Novel Therapeutic Approach using CXCR3 Blockade to Treat Immune Checkpoint Inhibitor-mediated Myocarditis

Huang, Y. V.; Lee, D.; Sun, Y.; Chou, H.; Xu, B.; Lin, Z.; Branche, C.; Bayer, A. L.; Waliany, S.; Neal, J.; Wakelee, H.; Witteles, R.; Nguyen, P.; Graves, E. E.; Alcaide, P.; Berry, G. J.; Wu, S. M.; Zhu, H.

doi:10.1101/2024.01.30.576279

bioRxiv · 10.1101/2024.01.30.576279

A Novel Therapeutic Approach using CXCR3 Blockade to Treat Immune Checkpoint Inhibitor-mediated Myocarditis

Huang, Y. V.·Lee, D.·Sun, Y.·Chou, H.·Xu, B.·Lin, Z.·Branche, C.·Bayer, A. L.·Waliany, S.·Neal, J.·Wakelee, H.·Witteles, R.·Nguyen, P.·Graves, E. E.·Alcaide, P.·Berry, G. J.·Wu, S. M.·Zhu, H.

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Abstract

BackgroundImmune checkpoint inhibitors (ICIs) are successful in treating many cancers but may cause immune-related adverse events. ICI-mediated myocarditis has a high fatality rate of up to 40%, with severe cardiovascular consequences. Targeted therapies for ICI myocarditis are currently lacking. MethodsWe used a genetic mouse model of PD-1 deletion (MRL/Pdcd1-/-) along with a novel drug-treated ICI myocarditis mouse model to recapitulate the disease phenotype. We performed single-cell RNA-sequencing (scRNAseq), single-cell T-cell receptor sequencing (scTCR-seq), and cellular indexing of transcriptomes and epitopes (CITE-seq) on immune cells isolated from MRL and MRL/Pdcd1-/- mice at serial timepoints. We assessed the impact of macrophage deletion in MRL/Pdcd1-/- mice, then inhibited CXC chemokine receptor 3 (CXCR3) in ICI-treated mice to assess therapeutic effect on myocarditis phenotype. Furthermore, we delineated functional effects of CXCR3 blockade on T-cell and macrophage interactions in a transwell assay. We then correlated the results in human single-cell multi-omics data from blood and heart biopsy data from patients with ICI myocarditis. ResultsSingle-cell multi-omics demonstrated expansion of CXCL9/10+CCR2+ macrophages and CXCR3hi CD8+ effector T-lymphocytes in the hearts of MRL/Pdcd1-/- mice correlating with onset of myocarditis development. Both depletion of CXCL9/10+CCR2+ macrophages and CXCR3 blockade respectively led to decreased CXCR3hiCD8+ T-cell infiltration into the heart and significantly improved survival. A transwell assay showed that selective blockade of CXCR3 and its ligand, CXCL10 decreased CD8+ T-cell migration towards macrophages, implicating this interaction in T-cell cardiotropism towards cardiac macrophages. Cardiac biopsies from patients with confirmed ICI myocarditis demonstrated infiltrating CXCR3+ lymphocytes and CXCL9+/CXCL10+ macrophages. Both mouse cardiac immune cells and patient peripheral blood immune cells revealed expanded TCRs correlating with CXCR3hi CD8+ T-cells in ICI myocarditis samples. ConclusionsThese findings bring forth the CXCR3-CXCL9/10 axis as an attractive therapeutic target for ICI myocarditis treatment, and more broadly, as a druggable pathway in cardiac inflammation.

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Huang, Y. V., Lee, D., Sun, Y., Chou, H., Xu, B., Lin, Z., Branche, C., Bayer, A. L., Waliany, S., Neal, J., Wakelee, H., Witteles, R., Nguyen, P., Graves, E. E., Alcaide, P., Berry, G. J., Wu, S. M., Zhu, H.. 2024-02-02. A Novel Therapeutic Approach using CXCR3 Blockade to Treat Immune Checkpoint Inhibitor-mediated Myocarditis. https://doi.org/10.1101/2024.01.30.576279

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