Early atherosclerosis depends upon responses by immune cells resident in the intimal aortic wall. Specifically, the healthy intima is thought to be populated by vascular dendritic cells (DC) that, during hypercholesterolemia, initiate atherosclerosis by being the first to accumulate cholesterol. Whether these cells remain key players in later stages of disease is unknown. Using murine lineage tracing models and gene expression profiling, we reveal that myeloid cells present in the intima of the aortic arch are not DCs but instead specialized aorta intima resident macrophages (Mac AIR ) that depend upon Csf-1 and sustained by local proliferation. Although Mac AIR comprise the earliest foam cells in plaques, their proliferation during plaque progression is limited. After months of hypercholesterolemia, their presence in plaque is overtaken by recruited monocytes, which induce Mac AIR -defining genes. These data redefine the lineage of intimal phagocytes and suggest that proliferation is insufficient to sustain generations of macrophages during plaque progression.

No abstract

Atherosclerotic plaque formation is driven by the continued expansion of cholesterol loaded foamy macrophages within the arterial intima. Foamy macrophages are primarily derived from newly recruited monocytes, but factors regulating monocyte specification toward foamy macrophage differentiation and prolonged survival in plaque remain poorly understood. We used trajectory analysis of integrated single cell RNA-seq data, along with a genome-wide CRISPR screening approach to identify Triggering Receptor Expressed on Myeloid Cells 2 (Trem2) as a candidate regulator for foamy macrophage specification. Loss of Trem2 led to a reduced ability of foamy macrophages to take up additional oxidized low density lipoprotein (LDL) cholesterol in vitro. Competitive chimera experiments showed that Trem2-deficient macrophages were less competent to form foamy macrophages when competed against Trem2-sufficient macrophages in vivo. In addition, myeloid specific conditional deletion of Trem2 resulted in a dramatic attenuation of plaque progression, even when targeted in established atherosclerotic lesions. This was independent of changes in circulating inflammatory cytokines, monocyte recruitment, or serum cholesterol levels, but due to a reduction in plaque macrophage proliferation and enhanced cell death. Mechanistically, we link Trem2-deficient macrophages with an inability for cells to sense cholesterol loading and failure to upregulate efflux molecules. Accumulation of cholesterol in the endoplasmic reticulum enhanced activation of the ER-stress response that increased susceptibility for cholesterol-toxicity and cell death in foamy Trem2-deficient macrophages. Overall, this study identifies Trem2 as a regulator of foamy macrophage differentiation, atherosclerotic plaque growth, and as a putative therapeutic target for future intervention studies.

Native freshwater mussels are a guild of benthic, filter feeding invertebrates that perform important ecological functions in rivers. Because of their long lifespans (30–50 years or longer), mussels are slow to respond to human‐induced alterations. Thus, development of sensitive indicators of mussel population responses to river conditions and management would be beneficial. Compared to marine species, estimation of vital rates (e.g. survival, growth) in freshwater mussels has received little attention. We placed passively integrated transponder tags on 578 mussels of four species (Amblema plicata, Cyclonaias pustulosa, Obliquaria reflexa, and Pleurobema sintoxia) in a well‐studied mussel assemblage in a side channel of the upper Mississippi River. Growth and survival of tagged mussels were assessed annually for 4 years across core (high density) and peripheral (low density) areas of the assemblage. Overall survival was highly variable, ranging from c. 15 to 90%, and was related to life history, habitat quality, and hydrologic events. Survival, which varied significantly among species and over time, was consistently higher in the dense and species‐rich core of the mussel assemblage, relative to the periphery because substrates were consistently more stable in the core of the mussel bed relative to the periphery. Substrate movement during low flows was an order of magnitude lower in the core relative to the periphery, and survival was inversely related to stability of river substrates. Patterns in habitat‐specific survival indicate source–sink population dynamics such that mussels in the core habitat provide recruitment to the periphery, but mussels in the periphery are subject to unsustainably low survival; additional studies to track the source of recruitment in the periphery are needed to test this hypothesis. Growth rate did not vary significantly between core and peripheral areas but did vary by species. Growth rate (proportional change per year) declined with age, and was similar at mean age for A. plicata (0.016 per year), P. sintoxia (0.015 per year), and C. pustulosa (0.013 per year), but much lower for O. reflexa (0.008 per year). Effective management decisions for mussels requires a better understanding of how vital rates govern populations and how they vary across a suite of physical and biological factors. Information on how population vital rates vary among species and over time gives managers another tool to understand how mussels may respond to management actions such as habitat restoration projects. Given the importance of substrate stability inferred from this study, management actions that maintain or increase substrate stability are likely to result in high quality mussel assemblages and may restore a valuable component of ecosystem function in this region.

No abstract