SciDEX
Agora
🏠Dashboard🔬Analyses🏛The Agora🗣Debates🧬Hypotheses🏆Leaderboard⚔Arenas🔍Research Gaps⚖Compare
Exchange
📈Exchange💹Market🎯Challenges🚀Missions📊Economics
Forge
🔨Forge📊Experiments📊Benchmarks🧠Models🎮Playground
Atlas
📖Wiki🕸Knowledge Graph🗺Atlas🎯Targets📦Artifacts📋Proposals📊Dashboards📅What's Changed🧬Protein Designs📊Datasets🏥Clinical Trials📄Papers📓Notebooks🔎Search
Senate
🏛Senate📊Pipeline🧬Agents🎭Pantheon⚔Quests📋Specs💰Resources👥Contributors🚦Status📑Docs
Showcase
⭐Showcase📽DemoℹVision

Defining the lineage of thermogenic perivascular adipose tissue.

Nature metabolism 2021
Open on PubMed

Brown adipose tissue can expend large amounts of energy, and therefore increasing its size or activity is a promising therapeutic approach to combat metabolic disease. In humans, major deposits of brown fat cells are found intimately associated with large blood vessels, corresponding to perivascular adipose tissue (PVAT). However, the cellular origins of PVAT are poorly understood. Here, we determine the identity of perivascular adipocyte progenitors in mice and humans. In mice, thoracic PVAT develops from a fibroblastic lineage, consisting of progenitor cells (Pdgfra<sup>+</sup>, Ly6a<sup>+</sup> and Pparg<sup>-</sup>) and preadipocytes (Pdgfra<sup>+</sup>, Ly6a<sup>+</sup> and Pparg<sup>+</sup>), which share transcriptional similarity with analogous cell types in white adipose tissue. Interestingly, the aortic adventitia of adult animals contains a population of adipogenic smooth muscle cells (Myh11<sup>+</sup>, Pdgfra<sup>-</sup> and Pparg<sup>+</sup>) that contribute to perivascular adipocyte formation. Similarly, human PVAT contains presumptive fibroblastic and smooth muscle-like adipocyte progenitor cells, as revealed by single-nucleus RNA sequencing. Together, these studies define distinct populations of progenitor cells for thermogenic PVAT, providing a foundation for developing strategies to augment brown fat activity.

72 Figures Extracted
Figure 1
Figure 1 PMC
(C, D) H&E staining (C) and mRNA levels of indicated genes (D) in thoracic aorta from C57/Bl6 mice housed at thermoneutrality or 4°C for one week ...
Figure 1
Figure 1 PMC
(H, I) H&E staining of thoracic aorta (H) and mRNA levels of indicated genes in eWAT and thoracic PVAT (I) from above mice. (n=6 mice per group; m...
Figure 1
Figure 1 PMC
(B) mRNA levels of indicated genes in thoracic PVAT, iBAT, and iWAT from C57/Bl6 mice. (n=5 mice per group; mean+/−SEM). One way ANOVA with Holm Sidak...
Figure 1
Figure 1 PMC
(G) Body weights of C57/Bl6 mice fed normal chow or western diet for 16 weeks (n=8 mice per group; mean+/−SEM). Two way-repeated measures ANOVA with H...
Figure 1
Figure 1 PMC
(E, F) H&E staining of thoracic aorta (E) and mRNA levels of indicated genes in aortic PVAT (F) from Ebf2WT and Ebf2ΔAdipoq mice (n=5 mice per gro...
Figure 1
Figure 1 PMC
Aortic PVAT expresses an EBF2-regulated classical brown fat program.
Figure 1
Figure 1 PMC
(A) H&E staining of thoracic aorta and iBAT from C57/Bl6 mice (scale bar, 100 μm) Representative images of n=5 experiments.
Figure 2
Figure 2 PMC
(H) UMAP showing expression of the progenitor marker gene Pi16 (left) and mRNA in-situ of hybridization of: (1) Pi16 (green) (middle); and (2) Clec11a...
Figure 2
Figure 2 PMC
(F) UMAP showing expression of preadipocyte marker genes and immunostaining of PPARg (red) and PDGFRa (green) in P3 thoracic aorta. Arrowheads show pr...
Figure 2
Figure 2 PMC
(A) H&E staining of retroperitoneum en bloc from E18 and P3 CD1 mice (scale bar, 517.6 μm). Representative of n=3 experiments.
Figure 2
Figure 2 PMC
(B) mRNA levels of indicated genes in thoracic aorta from E18 and P3 CD1 mice (n=3 biological replicates of pooled aortas; mean+/−SEM). Two sample, tw...
Figure 2
Figure 2 PMC
Lu: vessel lumen; DAPI (blue) was used to stain nuclei. Statistical Testing: NS p>0.05, **p≤0.01, ***p≤0.001, ****p≤0.0001.
Figure 2
Figure 2 PMC
(I) Model for thoracic aorta tissue organization at P3.
Figure 2
Figure 2 PMC
Identification of multiple fibroblast populations in developing aortic PVAT
Figure 2
Figure 2 PMC
(G) UMAP showing expression of intermediate cell genes and mRNA in-situ hybridization of Bace2 (red) in P3 thoracic aorta. Arrowheads indicate interme...
Figure 2
Figure 2 PMC
(E) UMAP showing expression of adipocyte marker genes (left) and immunostaining of PLIN1 (red) in P3 thoracic aorta. (scale bar, 543.5 μm). Representa...
Figure 2
Figure 2 PMC
(D) UMAP showing expression of smooth muscle marker genes (left) and immunostaining of PDGFRa (red) and MYH11 (green) in sections of E18 thoracic aort...
Figure 2
Figure 2 PMC
(C) UMAP of gene expression in 17,957 cells from P3 thoracic aorta of P3 CD1 mice.
Figure 3
Figure 3 PMC
(C) Pathway analysis of cluster-defining genes for each cell population. Cluster-defining genes are defined as significantly differentially expressed ...
Figure 3
Figure 3 PMC
Isolation of adipogenic fibroblasts from perinatal thoracic aorta
Figure 3
Figure 3 PMC
(E) Quantification of adipocyte differentiation (Bodipy+ area above threshold/nuclei) (Data pooled from n=5 separate experiments, n=5 biologically ind...
Figure 3
Figure 3 PMC
(D) Bodipy (lipid; green) and Hoechst (DNA; blue) staining of cell cultures following differentiation with adipogenic cocktail (Representative image o...
Figure 3
Figure 3 PMC
(A) FACS isolation of fibroblast and SMC populations. Live (FVS510-); Lin- (CD45-,CD31-,Ter119-) cells were gated as shown to isolate the following ce...
Figure 3
Figure 3 PMC
(B) Expression heatmaps of Seurat-generated cluster-defining genes mapped on to sorted-cell RNA-Seq results (n=3 biological replicates per group).
Figure 4
Figure 4 PMC
(C) Staining of GFP (green) and PLIN1 (red) in sections of thoracic aorta from Pdgfra-CreER(−); mTmG (control) and Pdgfra-CreER+; mTmG mice harvested ...
Figure 4
Figure 4 PMC
PVAT: Perivascular Adipose Tissue; Lu: vessel lumen; DAPI (blue) was used to stain nuclei.
Figure 4
Figure 4 PMC
(A) Schematic of cell types in perinatal aorta and corresponding marker genes. Created with BioRender.com.
Figure 4
Figure 4 PMC
Adipogenic fibroblasts are the major source of aortic adipocytes in neonates.
Figure 4
Figure 4 PMC
(B) Staining of PLIN1 (green) and RFP (red) in sections of P3 thoracic aorta from Myh11-Cre(−); tdTom (control) and Myh11-Cre+; tdTom reporter mice. (...
Figure 4
Figure 4 PMC
(D) Staining of GFP (green) in sections of P3 and adult thoracic aorta from Vipr2-Cre(−); mTmG and Vipr2-Cre+; mTmG mice. Arrowheads indicate GFP expr...
Figure 4
Figure 4 PMC
(E) Staining of GFP (green) in sections of P3 and adult thoracic aorta from Penk-Cre(−); mTmG and Penk-Cre+; mTmG+ mice. Yellow arrowheads show GFP+ f...
Figure 5
Figure 5 PMC
Comparative gene expression profiling of white and brown adipogenic cells
Figure 5
Figure 5 PMC
(C) Venn diagram of genes enriched in progenitor populations relative to preadipocyte populations from iWAT and PVAT. Pathway analysis of genes common...
Figure 5
Figure 5 PMC
(A) mRNA levels of indicated genes in differentiated adipocytes derived from the following cell types of P3 CD1 mice: iWAT preadipocytes, thoracic per...
Figure 5
Figure 5 PMC
Statistical Testing: NS p>0.05, **p≤0.01, ***p≤0.001, ****p≤0.0001.
Figure 5
Figure 5 PMC
(F) Model for conserved ontogeny of adipocyte development. Created with Biorender.com.
Figure 5
Figure 5 PMC
(E) Z-Score split heatmap of representative genes from GO analysis. Gene expression levels are calculated between cell types (i.e. PVAT Progenitors vs...
Figure 5
Figure 5 PMC
(D) Venn diagram of genes enriched in preadipocyte populations relative to progenitor populations. Pathway analysis of genes commonly enriched in both...
Figure 5
Figure 5 PMC
(B) Principal component analysis (PCA) of the top 500 differentially expressed genes in sorted cell RNA Seq. (n=3 biological replicates per cell type)...
Figure 6
Figure 6 PMC
(B) UMAPs showing expression of fibroblast marker genes.
Figure 6
Figure 6 PMC
(F) Pathway analysis of cluster-defining genes for each cell population. Cluster-defining genes were defined as significantly differentially expressed...
Figure 6
Figure 6 PMC
(E) Expression heatmap of Seurat-generated cluster defining genes mapped on to sorted-cell RNA-Seq results (n=3 biological replicates per group).
Figure 6
Figure 6 PMC
Identification of adipogenic smooth muscle cells (SMCs) in adult PVAT
Figure 6
Figure 6 PMC
(A) UMAP of gene expression in 6,753 cells from adult thoracic aorta of pooled 13-week-old male CD1 mice.
Figure 6
Figure 6 PMC
(C) UMAPs showing expression of SMC marker genes.
Figure 6
Figure 6 PMC
(D) FACS isolation of fibroblast and SMC populations. Live Lin(−) cells were gated to isolate the following cell populations: Intermediate Cells [PDGF...
Figure 7
Figure 7 PMC
(I) Model of thoracic aorta and associated tissue organization. Created with Biorender.com.
Figure 7
Figure 7 PMC
(C) Flow cytometry analysis of cell populations in stripped aorta or PVAT alone from adult mice (n=2 biological replicates of pooled aortas; mean+/−SE...
Figure 7
Figure 7 PMC
(D) UMAP of gene expression in 12,969 nuclei from adult human peri aortic PVAT (Subclustered to remove immune, endothelial, and mesothelial cells, n=3...
Figure 7
Figure 7 PMC
(E-G) UMAPs showing expression of adipocyte (E), Fibroblast (F) and SMC-related (G) marker genes in the dataset shown in (D).
Figure 7
Figure 7 PMC
Adipogenic activity of smooth muscle cells (SMCs) in adult PVAT
Figure 7
Figure 7 PMC
(A) Cell populations were FACS purified from adult aortas and induced to differentiate into adipocytes, followed by staining with Bodipy (lipid, green...
Figure 7
Figure 7 PMC
(B) Staining of RFP (red), PLIN1 (green), and DAPI (blue) in thoracic aorta from adult Myh11-Cre-; tdTom/+ and Myh11-Cre+; tdTom/+ mice treated with v...
Figure 7
Figure 7 PMC
(H) WikiPathways 2019 GO analysis of all genes differentially expressed (average logFC>0) in SMC-like cells vs. SMCs. Graph plots -log10 unadjusted...
Figure 7
Figure 7 PMC
Statistical Testing: NS p>0.05, **p≤0.01, ***p≤0.001, ****p≤0.0001.
Figure 8
Figure 8 PMC
(B-F) UMAPs showing expression of indicated genes for: smooth muscle cells (SMC) (B); Intermediate cells (C); Progenitors (D); Preadipocytes (E); and ...
Figure 8
Figure 8 PMC
(A) UMAPs of gene expression in cells from E18 and P3 thoracic aorta of pooled CD1 mice.
Figure 8
Figure 8 PMC
Single cell transcriptional profiling of aortas from E18 and P3 mice
Figure 8
Figure 8 PMC
(G) Expression dotplot of indicated genes for cell clusters from P3 aorta.
Figure 9
Figure 9 PMC
Purification of smooth muscle and fibroblast cells from aorta (Related to Fig. 3)
Figure 9
Figure 9 PMC
(A) FACS isolation of fibroblastic and smooth muscle cell (SMC) populations. Dissociated cells were gated on: (1) SSC-A and FSC-A to exclude debris; (...
Figure 10
Figure 10 PMC
Expression profiles for Cre-driver mouse lines (Related to Fig. 4)
Figure 10
Figure 10 PMC
(A) UMAPs showing expression of genes used for Cre and CreER driver mouse strains.
Figure 10
Figure 10 PMC
(B) Vipr2 expression levels in indicated cell types from sorted cell bulk RNAseq analysis. (One way ANOVA followed by two sided pairwise comparisons w...
Figure 11
Figure 11 PMC
Gene profiling analyses of iWAT vs. PVAT progenitors and preadipocytes (Related to Fig. 5)
Figure 11
Figure 11 PMC
(A) mRNA levels of indicated genes in differentiated primary adipocytes from: iWAT preadipocytes, aortic PVAT progenitors, aortic PVAT preadipocyte ce...
Figure 12
Figure 12 PMC
Purification and analysis of aorta-associated cells from adult animals (Related to Fig. 6)
Figure 12
Figure 12 PMC
(A) Expression dot plot of indicated genes in cell clusters from adult aorta. (B) mRNA in situ hybridization of Ly6a (green) in adult aorta. Arrowhead...
Figure 13
Figure 13 PMC
Analysis of aortic PVAT in adult mice (Related to Fig. 7)
Figure 13
Figure 13 PMC
(A) Staining for MYH11 (red) and PLIN (green) in aortas of Rosiglitazone (Rosi)-treated mice, scale bar 200 μm. Representative of n=1 experiment. Scal...
Figure 14
Figure 14 PMC
Single nucleus transcriptomic analyses of human PVAT (Related to Fig. 7)
Figure 14
Figure 14 PMC
(A) UMAP of gene expression in 18,758 nuclei from adult human peri-aortic PVAT (Full dataset, n=3 humans, integrated analysis). (B) Expression dotplot...
SciDEX — scidex.ai — Dashboard | Mission Control | Status | How it Works | GitHub