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Title GLOBAL SINGLE CELL SEQUENCING REVEALS EXTENSIVE CELL-CELL CROSSTALK AND SPECIFIC REGULATION OF MACROPHAGE INFLAMMATORY POLARISATION BY CARDIAC FIBROBLASTS FOLLOWING MYOCARDIAL INJURY |
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Type Free Paper Session 1 |
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Theme ACC Asia & SCS 32nd Annual Scientific Meeting |
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Topic Basic Science |
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Main Author Matthew Ackers-Johnson1 2 |
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Presenting Author Matthew Ackers-Johnson1 2 |
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Co-Author Motakis Efthymios1 2 Justus Stenzig3 Rongrong Zhao1 2 Zenia Tiang2 Wilson Tan2 Tuan Luu1 Peter Li1 Roger Foo1 2 |
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Department / Institution / Country Cardiovascular Research Institute / National University of Singapore / Singapore1 Human Genetics / Genome Institute of Singapore / Singapore2 Institut für Experimentelle Pharmakologie und Toxikologie / Universitätsklinikum Hamburg-Eppendorf / Germany (Deutschland)3 |
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Objective(s) Cardiac function and injury responses depend upon complex interactions between multiple specialised cell types of various origin, including infiltrating inflammatory immune cells, which are increasingly understood to have critical roles in disease. Research is however limited by inherent difficulties in identifying, isolating and characterising relevant cellular populations. We sought to apply an unbiased transcriptomic approach to characterise global cardiac non-myocyte populations and map intercellular cross-talk, in order to identify novel disease-associated cell markers and interactions, which could represent targets for future therapies. |
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Material and Method A flow cytometry single-cell RNA sequencing “FACS-seq” approach was developed to characterise non-myocyte populations from murine myocardium. Over 2000 cells were collected one week after surgically induced myocardial infarction (disease group) or sham surgery (control group). Single cell transcriptomic profiles were generated, and integrated with published datasets to create global cardiac intercellular interaction maps. Validation was performed using mouse and human histological sections and cardiac primary cell culture models. |
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Result(s) All major cardiac cell types were identified at relative proportions in line with recent reports, and new putative cell-type and disease-associated marker genes were revealed and validated. Global interaction maps revealed a striking increase in specific cross-talk between resident cardiac fibroblasts and macrophages in the disease environment. A unique regulatory function for cardiac fibroblasts in suppressing pro-inflammatory macrophage activation and driving macrophages towards an M2, anti-inflammatory polarised phenotype, was confirmed using in-vitro mouse and human cell culture models. |
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Conclusion We have developed a FACS-Seq procedure to enable unbiased, marker-independent analysis of myocardial cell populations. This technique revealed extensive cell-cell crosstalk and specific regulation of macrophage inflammatory polarisation by cardiac fibroblasts following cardiac injury. We anticipate that further investigation into this immunomodulatory mechanism will lead to the elucidation of new targets for therapeutic intervention. |