RNA sequencing data of Ing4;Ing5 double knock-out embryonic mouse hearts

Inhibitor of growth 4 and 5 (ING4 and ING5) are chromatin-binding proteins in the KAT6A, KAT6B and KAT7 histone acetyltransferase protein complexes. Heterozygous mutations in the KAT6A or KAT6B gene cause human disorders with cardiac defects, but the contribution of their chromatin adaptor proteins to development is unknown. We found that Ing5-/- mice had isolated cardiac ventricular septal defects. ING4 and ING5 are structurally similar proteins, suggesting a degree of redundancy. Combined loss of ING4 and ING5 caused developmental arrest at embryonic day E8.5, loss of histone H3 lysine 14 acetylation (H3K14ac), a reduction in H3K23ac levels and disruption of developmental gene expression in Ing4-/-Ing5-/- compared to control embryos. E12.5 Ing4+/- Ing5-/- hearts showed a paucity of epicardial cells and epicardium-derived cells, failure of myocardium compaction and coronary vasculature defects, accompanied by a reduction in the expression of epicardium genes compared to control hearts. In addition, a reduction in the expression of genes required for cell adhesion, both in whole E8.75 Ing4-/-Ing5-/- embryos and in E10.5 Ing4+/- Ing5-/- hearts compared to controls was observed. In vitro assessment of fibroblast and proepicardium explants revealed cell spreading and outgrowth defects. Our findings suggest that ING4 and ING5 are essential for heart development and promote epicardium and epicardium-derived cell fates and mutation of the human ING5 gene as a possible cause of isolated ventricular septal defects. RNA seqencing data was generated from embryonic mouse hearts taken at embryonic day 8.25 or 8.75 (E8.25 or E8.75). The embryos were either wild type (WT) or Ing4-/-;Ing5-/- double knock-outs. Each genotype group has n=5 biological replicates. Differential expression analyses were conducted between the genotype groups.生长抑制因子4与5（Inhibitor of Growth 4 and 5，ING4、ING5）是组蛋白乙酰转移酶KAT6A、KAT6B及KAT7蛋白复合物中的染色质结合蛋白。KAT6A或KAT6B基因的杂合突变会引发伴有心脏缺陷的人类疾病，但其染色质衔接蛋白对发育过程的贡献尚不明确。本研究发现，Ing5基因敲除纯合子（Ing5-/-）小鼠可出现单纯性室间隔缺损。ING4与ING5为结构相似的蛋白，提示二者存在一定程度的功能冗余。与对照胚胎相比，Ing4-/-Ing5-/-双敲除胚胎中，ING4与ING5的同时缺失会导致胚胎发育阻滞于胚胎第8.5天（E8.5），出现组蛋白H3赖氨酸14乙酰化（H3K14ac）丧失、H3K23乙酰化（H3K23ac）水平降低以及发育相关基因表达紊乱。E12.5时期的Ing4+/-;Ing5-/-小鼠心脏表现为心外膜细胞及心外膜来源细胞匮乏、心肌致密化不全以及冠状动脉血管缺陷，同时伴随心外膜相关基因表达水平下调，与对照心脏相比差异显著。此外，与对照组相比，全胚胎E8.75时期的Ing4-/-Ing5-/-胚胎以及E10.5时期的Ing4+/-;Ing5-/-心脏中，细胞黏附相关基因的表达均出现下调。对成纤维细胞及心外膜前体组织外植体的体外实验显示，细胞铺展与向外生长存在缺陷。本研究结果表明，ING4与ING5对心脏发育至关重要，可促进心外膜及心外膜来源细胞的命运决定，且人类ING5基因的突变可能是单纯性室间隔缺损的潜在致病诱因。本研究的RNA测序数据来源于胚胎第8.25天或8.75天（E8.25、E8.75）的小鼠胚胎心脏组织，受试胚胎分为野生型（wild type，WT）及Ing4-/-;Ing5-/-双敲除型两组，每个基因型组设置5个生物学重复样本（n=5），并对两组基因型样本开展了差异表达分析。