Deficiency of NONO is associated with impaired cardiac function and fibrosis in mice
Deficiency of NONO is associated with impaired cardiac function and fibrosis in mice
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NONO 缺乏与小鼠心脏功能受损和纤维化有关
DOI：
10.1016/j.yjmcc.2019.10.004
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发表时间：
2019-12-01
影响因子：
5
通讯作者：
Zhang, Yun
中科院分区：
文献类型：
--
作者：
Xu, Xingli;Jiang, Hong;Zhang, Yun
非POU结构域八聚体结合蛋白（non-POU-domain-containing octamer-binding protein，NONO）是多功能果蝇行为/人类剪接（DBHS）家族的成员，在调节糖脂代谢、昼夜节律、细胞分裂、胶原形成和纤维化等方面发挥重要作用。NONO的功能障碍变体被描述为男性先天性心脏缺陷的原因。但NONO缺乏对心室功能和心肌纤维化的影响及其机制尚不清楚。在本研究中，我们的目的是揭示整体表型，心脏功能和成纤维细胞在NONO敲除（NONO KO）小鼠与野生型（WT）雄性同窝仔相比。结果显示，NONOgt/0小鼠的出生率在断奶时远低于其WT雄性同窝出生小鼠。NONOgt/0小鼠的体重比WT同窝雄性小鼠的体重低19%（27.2 +/- 1.49 g对22.01 +/- 1.20 g，P < .001）。NONO KO小鼠从出生到一年后表现出持续较高的死亡率（P <0.05）。与野生型小鼠相比，（142.0 +/- 8.7 mg vs. 179.0 +/- 10.4 mg，P <0.001），心脏重量与体重比（HW/BW）相似，E/A比值较高（1.80 +/- 0.47 vs. 1.44 +/- 0.26，P <0.05），左心室舒张末期直径（LVEDd）显著降低（2.72 +/- 0.51 mm vs.3.54 +/- 0.43 mm，P <0.001）。我们还发现了过量的基质沉积在体内。在体外，NONO缺乏导致成纤维细胞过度增殖，而迁移受到抑制，这将诱导胶原成熟和沉积。相反，NONO的过表达抑制成纤维细胞增殖并增加迁移，从而减少胶原沉积。心脏成纤维细胞的RNA-seq进一步表明，NONO缺陷上调细胞周期调节因子，包括细胞周期蛋白B2，起源识别复合物1（ORC 1）和细胞分裂周期6（CDC 6），同时下调迁移调节因子，包括肌球蛋白，整合素和凝血因子II。NONO的过表达进一步验证了这些指标的作用。总之，我们的研究表明NONO缺乏与小鼠心脏缺陷的发生有关。NONO KO小鼠心脏成纤维细胞的过度增殖和胶原的过度分泌可能是导致心脏缺陷的原因。
Non-POU-domain-containing octamer-binding protein (NONO), a component of multifunctional Drosophila behavior/human splicing (DBHS) family, plays an important role in regulating glucose and fat metabolism, circadian cycles, cell division, collagen formation and fibrosis. Dysfunctional variants of NONO have been described as the cause of congenital heart defects in males. However, the effects of NONO deficiency on the ventricular function and cardiac fibrosis as well as the related mechanisms are not clear. In the present study, we aimed to reveal the overall phenotypes, cardiac function and fibroblasts in NONO knockout (NONO KO) mice compared with the wild-type (WT) male littermates. The results showed that the birth rate of NONOgt/0 mice was much lower than their WT male littermates at the time of weaning. The body weight of NONOgt/0 mice was 19% lower than that of WT male littermates (27.2 +/- 1.49 g vs. 22.01 +/- 1.20 g, P < .001). NONO KO mice exhibited continuous higher mortality from birth to a year later (P < .05). Compared with those in the WT mice, the heart weight was lower(142.0 +/- 8.7 mg vs. 179.0 +/- 10.4 mg, P < .001), the heart weight to body weight ratio (HW/BW) was similar, the E/A ratio was higher (1.80 +/- 0.47 vs. 1.44 +/- 0.26, P < .05), and the left ventricular end diastolic diameter (LVEDd) was significantly lower (2.72 +/- 0.51 mm vs.3.54 +/- 0.43 mm, P < .001) in the NONO KO mice. We also found excessive matrix deposition in vivo. In vitro, NONO deficiency led to fibroblasts hyperproliferation, while migration was inhibited, which would induce collagen maturation and deposition. Conversely, overexpression of NONO inhibited fibroblasts proliferation and increased migration which reduced collagen deposition. RNA-seq of cardiac fibroblasts further indicated that NONO deficiency upregulated the cell cycle regulators, which included cyclin B2, the origin recognition complex 1 (ORC1) and cell division cycle 6 (CDC6), while downregulated the migration regulators, which included myosins, integrin and coagulation factor II. Overexpression of NONO further verified the effects of these indicators. In conclusion, our study demonstrated that NONO deficiency was associated with developing heart defects in mice. Hyperproliferation of cardiac fibroblasts with dramatically excessive collagen secretion might be the cause of heart defects of NONO KO mice.