Human pericardial macrophages suppress cardiac fibrosis through cystatin C signaling after myocardial infarction
Ali Fatehi Hassanabad, Sarthak Sinha, Arzina Jaffer, Darrell Belke, Nicole L. Rosin, Elodie Labit, Daniel Young, Friederike I. Schoettler, Keerthana Chockalingam, Benjamin Haeyul Lee, Jameson A. Dundas, Emilie de Chantal, Carmina A. Isidoro, Alexander Tam, Hanjoo B. Shim, Anna N. Zarzycki, Afshin Derakhshani, Elisabeth Gorgiogianni, Jeannine D. Turnbull, Antoine Dufour, Shalina S. Ousman, Jeff A. Biernaskie, Paul W.M. Fedak, Justin F. Deniset
Ali Fatehi Hassanabad, Sarthak Sinha, Arzina Jaffer, Darrell Belke, Nicole L. Rosin, Elodie Labit, Daniel Young, Friederike I. Schoettler, Keerthana Chockalingam, Benjamin Haeyul Lee, Jameson A. Dundas, Emilie de Chantal, Carmina A. Isidoro, Alexander Tam, Hanjoo B. Shim, Anna N. Zarzycki, Afshin Derakhshani, Elisabeth Gorgiogianni, Jeannine D. Turnbull, Antoine Dufour, Shalina S. Ousman, Jeff A. Biernaskie, Paul W.M. Fedak, Justin F. Deniset
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Research Article Cardiology Inflammation

Human pericardial macrophages suppress cardiac fibrosis through cystatin C signaling after myocardial infarction

Abstract

The pericardium plays an important homeostatic role for the neighboring heart, providing both lubrication and structural support. In vivo models have further identified a protective role for the pericardium in modulating cardiac remodeling following myocardial infarction, possibly through the actions of tissue-resident pericardial macrophages. Using patient-derived pericardial samples, we establish that human pericardial immune cells directly inhibit cardiac fibroblast fibrotic activity, and this action is dampened following myocardial infarction. Using single-cell RNA sequencing of patient pericardial fluid cells, we identify two pericardial macrophage subsets that are uniquely altered in response to myocardial infarction, which contributes to a shift in their effector molecule expression profiles. We confirm that fibronectin-expressing human pericardial macrophages are the primary driver of the pericardial antifibrotic actions through the release of cystatin C. Finally, we establish cystatin C as a myeloid cell–derived cardioprotective effector molecule in an in vivo model of myocardial infarction. Collectively, we uncover a molecular mechanism of the local immune environment that regulates cardiac remodeling after myocardial infarction.

Authors

Ali Fatehi Hassanabad, Sarthak Sinha, Arzina Jaffer, Darrell Belke, Nicole L. Rosin, Elodie Labit, Daniel Young, Friederike I. Schoettler, Keerthana Chockalingam, Benjamin Haeyul Lee, Jameson A. Dundas, Emilie de Chantal, Carmina A. Isidoro, Alexander Tam, Hanjoo B. Shim, Anna N. Zarzycki, Afshin Derakhshani, Elisabeth Gorgiogianni, Jeannine D. Turnbull, Antoine Dufour, Shalina S. Ousman, Jeff A. Biernaskie, Paul W.M. Fedak, Justin F. Deniset

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Figure 6

Myeloid-derived Cst3 alters cardiac fibrosis.

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Myeloid-derived Cst3 alters cardiac fibrosis. (A) Representative flow c...
(A) Representative flow cytometry analysis of mouse GPCM expression of FN1, TIM4, and LYVE1. Representative of n = 4. (B) RNA-seq quantification of top-expressed genes by sorted mouse GPCMs. n = 4. (C) qPCR quantification of Cst3 mRNA expression in pericardial macrophages, cardiac macrophages, and Ly6Chi monocytes from WT (Cst3fl/fl) and Cst3ΔLyz2 mice (left), and ELISA quantification of serum (middle) and pericardial lavage (right) CST3 levels from WT and Cst3ΔLyz2 mice. n = 3 for qPCR; n = 8 for serum samples; n = 6 and 4 for pericardial lavage samples from WT and Cst3ΔLyz2 mice, respectively. *P < 0.05, **P < 0.01, unpaired 2-tailed t test for ELISA; ****P < 0.0001, 1-way ANOVA with Tukey’s multiple-comparison test for qPCR. (D) Immunohistochemistry staining and quantification for α-smooth muscle actin (α-SMA) and collagen I (Col I) in the infarct and border zones from WT (Cst3fl/fl) and Cst3ΔLyz2 mice at 7 days after MI. Scale bars: 1,000 μm. Data are represented as mean ± SEM; n = 6 and 8 for WT and Cst3ΔLyz2 mice, respectively. Unpaired 2-tailed t test, non-parametric (Mann-Whitney) for α-SMA and parametric (Welch’s correction) for Col I. (E) Schematic of experimental timeline for cardiac function and fibrosis analysis at 28 days after MI. (F) Representative confocal composite stitch images of the LV and border zone (BZ) with Picrosirius red (PSR) staining and quantification of total LV scar size and BZ fibrosis indicated by PSR staining in cardiac cross sections at 28 days after MI for WT and Cst3ΔLyz2 mice. Scale bars: 1,000 μm. Data are represented as mean ± SEM; n = 8 and 10 for WT and Cst3ΔLyz2 mice, respectively. *P < 0.05, 2-tailed t test. (G) LV functional parameters (end systolic pressure volume relationship [ESPVR], preload recruitable stroke work [PRSW], and end diastolic pressure volume relationship [EDPVR]) at 28 days after MI for WT and Cst3ΔLyz2 mice measured by pressure-volume assessment. Data are represented as mean ± SEM; n = 14 for WT and n = 15 for Cst3ΔLyz2 mice. *P < 0.05, unpaired 2-tailed t test.