Lu Yang, Wu Qiongfeng, Liao Jie, Zhang Shaoshao, Lu Kai, Yang Shuaitao, Wu Yuwei, Dong Qian, Yuan Jing, Zhao Ning, Du Yimei
Department of Cardiology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
Research Center of Ion Channelopathy, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
Ann Transl Med. 2021 Sep;9(18):1401. doi: 10.21037/atm-21-2913.
Dilated cardiomyopathy (DCM) is a common cause of heart failure. Cardiac remodeling is the main pathological change in DCM, yet the molecular mechanism is still unclear. Therefore, the present study aims to find potential crucial genes and regulators through bulk and single-cell transcriptomic analysis.
Three microarray datasets of DCM (GSE57338, GSE42955, GSE79962) were chosen from gene expression omnibus (GEO) to analyze the differentially expressed genes (DEGs). LASSO regression, SVM-RFE, and PPI network methods were then carried out to identify key genes. Another dataset (GSE116250) was used to validate these findings. To further identify DCM-associated specific cell types, transcription factors, and cell-cell interaction networks, GSEA, SCENIC, and CellPhoneDB were conducted on public datasets for single-cell RNA sequencing analysis of DCM (GSE109816 and GSE121893). Finally, reverse transcription-polymerase chain reaction (RT-PCR), western blot, and immunohistochemical were performed to validate DPT expression in fibroblasts and DCM.
There were 281 DEGs between DCM and non-failing donors. CCL5 and DPT were identified to be key genes and both genes had a 0.844 area under the receiver operating curve (AUC) in the validation dataset. Further single-cell sequencing analysis revealed three main findings: (I) DPT was mainly expressed in fibroblasts and was significantly upregulated in DCM fibroblasts; (II) DPT fibroblasts were involved in the organization of the extracellular matrix (ECM) and collagen fibrils and were regulated by the transcription factor STAT3; and (III) DPT fibroblasts had high interactions with endothelial cells through including Ephrin-Eph, ACKR-CXCL, and JAG-NOTCH signal pathways. RT-PCR, western blot, and immunohistochemical confirmed that DPT was highly expressed and co-localized with Vimentin and p-STAT3 in DCM patients. STAT3 inhibitor S3I-201 reduced the expression of DPT in mouse cardiac fibroblasts.
DPT could be used as a diagnostic marker and therapeutic target of DCM. DPT fibroblasts could be a novel regulator of the cardiac remodeling process in DCM.
扩张型心肌病（DCM）是心力衰竭的常见原因。心脏重塑是DCM的主要病理变化，但其分子机制仍不清楚。因此，本研究旨在通过批量和单细胞转录组分析寻找潜在的关键基因和调节因子。
从基因表达综合数据库（GEO）中选取三个DCM的微阵列数据集（GSE57338、GSE42955、GSE79962）来分析差异表达基因（DEG）。然后采用套索回归、支持向量机递归特征消除（SVM-RFE）和蛋白质-蛋白质相互作用（PPI）网络方法来鉴定关键基因。使用另一个数据集（GSE116250）来验证这些发现。为了进一步鉴定与DCM相关的特定细胞类型、转录因子和细胞-细胞相互作用网络，对DCM的单细胞RNA测序分析的公共数据集（GSE109816和GSE121893）进行基因集富集分析（GSEA）、单细胞调控网络推断和聚类（SCENIC）以及细胞间通讯分析工具（CellPhoneDB）。最后，进行逆转录-聚合酶链反应（RT-PCR）、蛋白质免疫印迹和免疫组织化学来验证DPT在成纤维细胞和DCM中的表达。
DCM与非衰竭供体之间有281个DEG。CCL5和DPT被鉴定为关键基因，在验证数据集中这两个基因的受试者工作特征曲线下面积（AUC）均为0.844。进一步的单细胞测序分析揭示了三个主要发现：（I）DPT主要在成纤维细胞中表达，在DCM成纤维细胞中显著上调；（II）DPT成纤维细胞参与细胞外基质（ECM）和胶原纤维的组织，并受转录因子STAT3调控；（III）DPT成纤维细胞通过Ephrin-Eph、ACKR-CXCL和JAG-NOTCH信号通路与内皮细胞有高度相互作用。RT-PCR、蛋白质免疫印迹和免疫组织化学证实DPT在DCM患者中高表达，并与波形蛋白和磷酸化STAT3共定位。STAT3抑制剂S3I-201降低了小鼠心脏成纤维细胞中DPT的表达。
DPT可作为DCM的诊断标志物和治疗靶点。DPT成纤维细胞可能是DCM心脏重塑过程中的一种新型调节因子。
