Bacterial infections shape cardiac macrophages’ response to ischemia

Background: Patients with bacterial infections are at increased risk for subsequent cardiovascular events. Whether infections’ effects on innate immune cells within the cardiovascular system influence subsequent pathologies remains unclear. Here, we explore cardiac myeloid cells’ chronic adaptations to a preceding bacterial insult and implications for subsequent myocardial ischemia. Methods: We used various flow cytometry protocols to assess cardiac immune cells, peripheral leukocytes and hematopoietic stem and progenitor cells in bone marrow and spleen. A genetic fate mapping model was used to determine cardiac macrophages’ origin after bacteremia. Cardiac leukocytes were analyzed using scRNAseq. Nanoparticle-mediated RNAi was used to target macrophages in-vivo. Results: Cardiac macrophage numbers increased sharply, and numerical changes alongside subset alterations persisted over time. Fate mapping pointed towards local origin as the primary macrophage source after infection. Profiling macrophage heterogeneity using scRNAseq, we identified two previously unknown subpopulations remaining after resolution of infection. Whereas heightened metabolic activity was one subset’s primary feature, the other displayed excessive chemotactic properties, which amplified cardiac leukocyte recruitment and inflammation after a subsequent ischemic injury. Targeting cardiac macrophages’ surplus inflammatory activity after infection using nanoparticle-enabled, macrophage-directed RNAi kept disproportionate subsequent ischemic inflammation at bay. Conclusions: Bacteremia induces long-lasting changes in the cardiovascular system’s innate immune cells’ composition. This may amplify myocardial inflammation after a subsequent ischemic injury. C57Bl6/J mice were subject to cecal ligation and puncture (CLP); 6 weeks later, animals were sacrificed and hearts were flushed and digested. From Single cell dispension, CD45 positive cells were isolated by fluorescence activated cell sorting and immediately prepped for single cell sequencing by 10x genomics workflow. Animals of the same age without treatment were used as a control.