Contribution of VEGF-B-induced endocardial endothelial cell lineage in physiological versus pathological cardiac hypertrophy.. Contribution of VEGF-B-induced endocardial endothelial cell lineage in physiological versus pathological cardiac hypertrophy.

ScRNA-seq was used to investigate the effects of autocrine versus paracrine VEGF-B signaling in the heart using transgenic mouse models. The paracrine model was further investigated in pregnancy-induced cardiac hypertrophy as well as in mice with ligation of the left anterior descending (LAD) coronary artery. Overall design: Murine hearts were isolated and digested followed by analysis of the stromovascular fraction (SVF) by scRNA-seq, or FAC sorting of the cardiac enodthelial cells (ECs) by using FITC-tagged Pecam1 antibody and analysis by scRNA-seq.

本研究采用单细胞RNA测序（single-cell RNA sequencing, scRNA-seq）技术，借助转基因小鼠模型，对比探究自分泌（autocrine）与旁分泌（paracrine）通路的血管内皮生长因子B（VEGF-B）信号在心脏中的作用。旁分泌VEGF-B信号模型还被进一步应用于妊娠诱导的心肌肥厚小鼠，以及接受左前降支（left anterior descending coronary artery, LAD）冠状动脉结扎术的小鼠中开展研究。实验设计概述：分离并消化小鼠心脏组织后，通过两种方案开展分析：其一为对血管基质组分（stromovascular fraction, SVF）进行单细胞RNA测序；其二为利用异硫氰酸荧光素（fluorescein isothiocyanate, FITC）标记的PECAM1抗体对心脏内皮细胞（cardiac endothelial cells, ECs）进行荧光激活细胞分选（fluorescence-activated cell sorting, FACS），随后对分选所得细胞实施单细胞RNA测序。