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. 2021 Feb;12(2):152-157.
doi: 10.1007/s13238-020-00788-6. Epub 2020 Nov 7.

Single-cell transcriptomics of cardiac progenitors reveals functional subpopulations and their cooperative crosstalk in cardiac repair

Lei Gao #  1 , Hongjie Zhang #  2 , Jingyi Cui  1 , Lijuan Pei  2 , Shiqi Huang  2 , Yaning Mao  1 , Zhongmin Liu  3 , Ke Wei  4 , Hongming Zhu  5
Affiliations

Single-cell transcriptomics of cardiac progenitors reveals functional subpopulations and their cooperative crosstalk in cardiac repair

Lei Gao et al. Protein Cell. 2021 Feb.
No abstract available

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Figures

Figure 1
Figure 1
scRNA-seq reveals two major subpopulations within CDCs. (A) Schematic of experimental strategy. CSps, cardiospheres; CDCs, cardiosphere-derived cells; scRNA-seq, single-cell RNA sequencing. (B) Uniform manifold approximation and projection (UMAP) plot revealed that CDCs contained 6 distinct clusters with removing cell cycle genes. No., number of cells. (C) Dot plot of marker genes for each cluster. Endo, endothelial cells; Ave. Exp., average expression. (D) Feature plot showing the distribution and expression of Ly6a in CDCs. Network diagram of ligand-receptor connections (E); comparison of total incoming/outgoing path weights (F); network diagram of angiogenesis-related ligand-receptor connectivity among Sca-1−, Sca-1+, and endothelial cells within CDCs (G). D, donor; R, recipient. Number of interactions are annotated on the edge. (H) Volcano plot illustrating differentially expressed genes (DEGs) between Sca-1− and Sca-1+ CDCs based on scRNA-seq dataset. Red, significantly upregulated genes; blue, significantly downregulated genes; gray, no significant difference. Fold change ≥ 2 and P < 0.05 were considered significant. (I) Feature plot showing the distribution and expression of Vegfa and Flt1 in CDCs. Sorting strategy (J) and representative gating strategy (K) to sort Sca-1− and Sca-1+ cells from CDCs by fluorescence-activated cell sorting (FACS). (L) Representative immunofluorescence images of sorted Sca-1− and Sca-1+ CDCs. Cells were stained with Sca-1 (red); nuclei were labelled with DAPI (blue). Representative images of 3 independent experiments are shown. Scale bar, 100 μm. (M) Angiogenic proteome analysis of CM from Sca-1− and Sca-1+ CDCs, respectively. CM, conditioned medium. (N) Quantification of the percentage of Flt1+ cells in Sca-1− and Sca-1+ CDCs as assessed by flow cytometry. Data are shown as mean ± SD from 4 biological replicates. *, significantly different from Sca-1− CDCs; **, P < 0.01. (O) Tube formation analysis of Sca-1− CDCs after treatment with indicated CM. Average tube length was quantified. Data are shown as mean ± SD from 3 independent experiments. *, significantly different from group a; **, P < 0.01. #, significantly different from group b; #, P < 0.05. (P) Tube formation analysis of Sca-1− and Sca-1+ CDCs. Data are shown as mean ± SD from 2 independent experiments. *, significantly different from Sca-1− CDCs; **, P < 0.01. (Q) Experimental strategy. (R) Representative bright field (BF) and CD31 immunohistochemical images. Scale bar, 100 μm. n = 6 mice per group. Data are shown as mean ± SD. *, significantly different from Sca-1− CDCs; **, P < 0.01
Figure 2
Figure 2
Sca-1+ CDCs have beneficial effects on the cardiac function after MI. (A) Representative images of echocardiography and echocardiographic measurements of EF. MI, myocardial infarction; LVEF, left ventricle ejection fraction. n = 7 mice per group at least. Data are shown as mean ± SD from 2 independent experiments. *, significantly different from control; **, P < 0.01. #, significantly different from MI; #, P < 0.05. (B) Representative images of Masson’s trichrome-stained heart sections. Scale bar, 1 mm. n = 4 mice per group. (C) Representative images of CD31 (red) expression in heart sections. Nuclei were labeled with DAPI (blue). Scale bar, 200 μm. n = 5 mice per group. Data are shown as mean ± SD. *, significantly different from MI; *, P < 0.05; **, P < 0.01. #, significantly different from Sca-1− CDCs; #, P < 0.05. (D) Volcano plot illustrating DEGs between Sca-1− and Sca-1+ CDCs based on RNA-seq dataset. Red, significantly upregulated genes; blue, significantly downregulated genes; gray, no significant difference. Fold change ≥ 2 and P < 0.05 were considered significant. (E) Top 5 GO terms of significantly upregulated DEGs in Sca-1+ CDCs. (F) Heatmap illustrating the expression of ligands within these significantly upregulated DEGs in Sca-1+ CDCs. Red, reported cardioprotective molecules. (G) Schematic diagram. There is a crosstalk between Sca-1+ and Sca-1− cells within CDCs mediated by Vegfa and Flt1. Sca-1+ CDCs transplantation improves the cardiac function after MI via secreting plenty of cardioprotective molecules
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References

    1. Cambier L, de Couto G, Ibrahim A, Echavez AK, Valle J, Liu W, Kreke M, Smith RR, Marban L, Marban E. Y RNA fragment in extracellular vesicles confers cardioprotection via modulation of IL-10 expression and secretion. EMBO Mol Med. 2017;9:337–352. doi: 10.15252/emmm.201606924. - DOI - PMC - PubMed
    1. Das S, Goldstone AB, Wang H, Farry J, D’Amato G, Paulsen MJ, Eskandari A, Hironaka CE, Phansalkar R, Sharma B, et al. A unique collateral artery development program promotes neonatal heart regeneration. Cell. 2019;176:1128–1142.e1118. doi: 10.1016/j.cell.2018.12.023. - DOI - PMC - PubMed
    1. DeLaughter DM, Bick AG, Wakimoto H, McKean D, Gorham JM, Kathiriya IS, Hinson JT, Homsy J, Gray J, Pu W, et al. Single-cell resolution of temporal gene expression during heart development. Dev Cell. 2016;39:480–490. doi: 10.1016/j.devcel.2016.10.001. - DOI - PMC - PubMed
    1. Ellison GM, Torella D, Dellegrottaglie S, Perez-Martinez C, Perez de Prado A, Vicinanza C, Purushothaman S, Galuppo V, Iaconetti C, Waring CD, et al. Endogenous cardiac stem cell activation by insulin-like growth factor-1/hepatocyte growth factor intracoronary injection fosters survival and regeneration of the infarcted pig heart. J Am Coll Cardiol. 2011;58:977–986. doi: 10.1016/j.jacc.2011.05.013. - DOI - PubMed
    1. Ibrahim AGE, Li C, Rogers R, Fournier M, Li L, Vaturi SD, Antes T, Sanchez L, Akhmerov A, Moseley JJ, et al. Augmenting canonical Wnt signalling in therapeutically inert cells converts them into therapeutically potent exosome factories. Nat Biomed Eng. 2019;3:695–705. doi: 10.1038/s41551-019-0448-6. - DOI - PMC - PubMed

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