Single-cell and spatial heterogeneity landscapes of mature epicardial cells

作者全名："Du, Jianlin; Yuan, Xin; Deng, Haijun; Huang, Rongzhong; Liu, Bin; Xiong, Tianhua; Long, Xianglin; Zhang, Ling; Li, Yingrui; She, Qiang"

作者地址："[Du, Jianlin; Liu, Bin; Xiong, Tianhua; Long, Xianglin; Li, Yingrui; She, Qiang] Chongqing Med Univ, Dept Cardiol, Affiliated Hosp 2, Chongqing 400010, Peoples R China; [Yuan, Xin] Chongqing Med Univ, Dept Nephrol, Affiliated Hosp 2, Chongqing 400010, Peoples R China; [Deng, Haijun] Chongqing Med Univ, Inst Viral Hepatitis, Key Lab Mol Biol Infect Dis, Minist Educ,Affiliated Hosp 2, Chongqing 400010, Peoples R China; [Huang, Rongzhong] Chongqing Med Univ, Precis Med Ctr, Affiliated Hosp 2, Chongqing 400010, Peoples R China; [Zhang, Ling] Chongqing Med Univ, Basic Med Res & Innovat Ctr Novel Target & Therap, Minist Educ, Affiliated Hosp 2, Chongqing 400010, Peoples R China"

通信作者："Li, YR; She, Q (通讯作者)，Chongqing Med Univ, Dept Cardiol, Affiliated Hosp 2, Chongqing 400010, Peoples R China.; Zhang, L (通讯作者)，Chongqing Med Univ, Basic Med Res & Innovat Ctr Novel Target & Therap, Minist Educ, Affiliated Hosp 2, Chongqing 400010, Peoples R China."

来源：JOURNAL OF PHARMACEUTICAL ANALYSIS

ESI学科分类：PHARMACOLOGY & TOXICOLOGY

WOS号：WOS:001083841000001

JCR分区：Q1

影响因子：6.1

年份：2023

卷号：13

期号：8

开始页：894

结束页：907

文献类型：Article

关键词：Epicardial cells; Gene markers; Single-cell sequencing; Spatial transcriptomics

摘要："Tbx18, Wt1, and Tcf21 have been identified as epicardial markers during the early embryonic stage. However, the gene markers of mature epicardial cells remain unclear. Single-cell transcriptomic analysis was performed with the Seurat, Monocle, and CellphoneDB packages in R software with standard procedures. Spatial transcriptomics was performed on chilled Visium Tissue Optimization Slides (10x Genomics) and Visium Spatial Gene Expression Slides (10x Genomics). Spatial transcriptomics analysis was performed with Space Ranger software and R software. Immunofluorescence, whole-mount RNA in situ hybridization and X-gal staining were performed to validate the analysis results. Spatial transcriptomics analysis revealed distinct transcriptional profiles and functions between epicardial tissue and non-epicardial tissue. Several gene markers specific to postnatal epicardial tissue were identified, including Msln, C3, Efemp1, and Upk3b. Single-cell transcriptomic analysis revealed that cardiac cells from wildtype mouse hearts (from embryonic day 9.5 to postnatal day 9) could be categorized into six major cell types, which included epicardial cells. Throughout epicardial development, Wt1, Tbx18, and Upk3b were consistently expressed, whereas genes including Msln, C3, and Efemp1 exhibited increased expression during the mature stages of development. Pseudotime analysis further revealed two epicardial cell fates during maturation. Moreover, Upk3b, Msln, Efemp1, and C3 positive epicardial cells were enriched in extracellular matrix signaling. Our results suggested Upk3b, Efemp1, Msln, C3, and other genes were mature epicardium markers. Extracellular matrix signaling was found to play a critical role in the mature epicardium, thus suggesting potential therapeutic targets for heart regeneration in future clinical practice. (c) 2023 The Author(s). Published by Elsevier B.V. on behalf of Xi'an Jiaotong University. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/)."

基金机构："Supercomputing Center of Chongqing Medical University; National Natural Science Foundation of China [82270281]; Chongqing Medical University Program for Youth Innovation in Future Medicine [W0133]; Senior Medical Talents Program of Chongqing for Young and Middle-aged, China; Post-doctoral Research Funding of the Second Affiliated Hospital of Chongqing Medical University, China [rsc-postdoctor114]; Kuanren Talents Program of the Second Affiliated Hospital of Chongqing Medical University, China [kryc-gg-2102]"

基金资助正文："Tbx18: Cre mice were donated from Professor Sylvia M. Evans. We thank Dr. Andreas Kispert from Hannover Medical School (MHH) for providing probe plasmids of Tbx18. The computational work in this paper received partial support from the Supercomputing Center of Chongqing Medical University. This work was supported by grants from the National Natural Science Foundation of China (Grant No.: 82270281), Chongqing Medical University Program for Youth Innovation in Future Medicine (Grant No.: W0133), Senior Medical Talents Program of Chongqing for Young and Middle-aged, China (Program No.: JianlinDu [2022]), Post-doctoral Research Funding of the Second Affiliated Hospital of Chongqing Medical University, China (Grant No.: rsc-postdoctor114), and Kuanren Talents Program of the Second Affiliated Hospital of Chongqing Medical University, China (Program No.: kryc-gg-2102)."