Mann-Whitney test, ????p?< 0.0001. (D) Representative immunofluorescence and RNAscope images of murine livers following acute CCl4 administration: NGFR/(RNAscope) (red), (RNAscope) (green), DAPI (blue). to Figures 4 and 6 This table provides the list of genes associated with each module of differentially expressed genes over the central-associated HSC pseudotemporal trajectory in homeostatic and fibrotic murine hepatic mesenchyme (Figures 4E and 4F), the central-associated HSC pseudotemporal trajectory in acute-injury murine hepatic mesenchyme (Figures 6C and 6D), and the portal-associated HSC pseudotemporal trajectory in acute-injury murine hepatic mesenchyme (Figures 6E and 6F), as well as their associated GO terms. It also provides the list of transcription factor regulons differentially expressed over the central-associated HSC pseudotemporal trajectory in homeostatic and fibrotic murine hepatic mesenchyme and the central-associated quiescent to activated HSC pseudotemporal trajectory in acute-injury murine hepatic mesenchyme (Figures S9C and S9D). mmc4.xlsx (4.8M) GUID:?C964742F-ADE5-4647-9CF3-C42F3F840AD8 Table S4. Antibodies Used for Immunofluorescence, Related to STAR Methods This table provides a list of commercial antibodies and conditions used in this study (STAR Methods). mmc5.xlsx (10K) GUID:?B026354E-EEFA-41C4-B0D4-9D0B4BFCE549 Document S2. Article plus Supplemental Information mmc6.pdf (22M) GUID:?1F82B6FE-43BF-44A3-87AC-E564C74F2A21 Data Availability StatementAll mouse mesenchymal data Senicapoc (ICA-17043) is deposited in the Gene Expression Senicapoc (ICA-17043) Omnibus. The accession number for the data is GEO: “type”:”entrez-geo”,”attrs”:”text”:”GSE137720″,”term_id”:”137720″GSE137720. All human mesenchymal data, as well as mouse leucocyte data, is available from the Gene Expression Omnibus (GEO: “type”:”entrez-geo”,”attrs”:”text”:”GSE136103″,”term_id”:”136103″GSE136103). R markdown scripts enabling the Rabbit Polyclonal to TESK1 main steps of the analysis are available from the Lead Contact upon reasonable request. Additional Resources Our uninjured and 6?week CCl4 expression data is freely available for user-friendly interactive browsing online: http://livermesenchyme.hendersonlab.mvm.ed.ac.uk Summary Iterative liver injury results in progressive fibrosis disrupting hepatic architecture, regeneration potential, and liver function. Hepatic stellate cells (HSCs) are a major source of pathological matrix Senicapoc (ICA-17043) during fibrosis and are thought to be a functionally homogeneous population. Here, we use single-cell RNA sequencing to deconvolve the hepatic mesenchyme in healthy and fibrotic mouse liver, revealing spatial zonation of HSCs across the hepatic lobule. Furthermore, we show that HSCs partition into topographically diametric lobule regions, designated portal vein-associated HSCs (PaHSCs) and central vein-associated HSCs (CaHSCs). Importantly we uncover functional zonation, identifying CaHSCs as the dominant pathogenic collagen-producing cells in a mouse model of centrilobular fibrosis. Finally, we identify LPAR1 as a therapeutic target on collagen-producing CaHSCs, demonstrating that blockade of LPAR1 inhibits liver fibrosis in a rodent NASH model. Taken together, our work illustrates the power of single-cell transcriptomics to resolve the key collagen-producing cells driving liver fibrosis with high precision. R package (Camp et?al., 2017) to visualize coordinately expressed gene groups across the transcriptomic landscape (Figure?S2F). We identified three metagene signatures, denoted as ACC, that strongly define the subpopulations (Table S2). Signature A, enriched for gene ontology (GO) terms relating to extracellular structure organization, defined both FBs and VSMCs mesenchymal subpopulations. Signature B defined the HSCs subpopulation and was enriched for terms including retinoid metabolic process and antigen processing and presentation. Signature C defined VSMCs exclusively and was enriched for terms such as actin filament-based processes (Figures 1F and S2F). Using a single-cell approach also allowed us to interrogate traditional hepatic mesenchymal markers at high resolution. We found that certain historic HSC markers, such as and expression was negligible in our dataset. We confirmed and as specific markers for HSCs within the hepatic mesenchyme (Lua et?al., 2016, Mederacke et?al., 2013), and displayed a spectrum of expression across the HSC population. expression was confined to the FB and HSC subpopulations as Senicapoc (ICA-17043) opposed to and and 29 genes associated with and including and (Figure?2A; Table S1). Supervised clustering using this signature?allowed us to.