Dlk1 maintains adult mice long-term HSCs by activating Notch signaling to restrict mitochondrial metabolism.
Dlk1 maintains adult mice long-term HSCs by activating Notch signaling to restrict mitochondrial metabolism.
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Dlk1通过激活Notch信号限制线粒体代谢来维持成年小鼠的长期HSC
DOI：
10.1186/s40164-022-00369-9
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发表时间：
2023-01-18
影响因子：
10.9
通讯作者：
Qian, Pengxu
中科院分区：
文献类型：
--
作者：
Huang, Deyu;Han, Yingli;Tang, Tian;Yang, Lin;Jiang, Penglei;Qian, Wenchang;Zhang, Zhaoru;Qian, Xinyue;Zeng, Xin;Qian, Pengxu
成体造血干细胞（HSC）的稳态对于维持终生造血至关重要。然而，成人造血干细胞如何协调其稳态仍不完全清楚。印迹基因 Dlk1 已被证明在小鼠胚胎造血和干细胞调节中发挥关键作用，但其在成人 HSC 中的生理作用尚不清楚。我们对 Dlk1 进行了基因表达分析，并通过将 Mx1 cre 小鼠与 Dlkflox/flox 小鼠杂交构建了条件 Dlk1 敲除 (KO) 小鼠。采用Western blot和定量PCR检测Dlk1 KO效率。采用流式细胞术研究 Dlk1 KO 对原代小鼠 HSC、祖细胞和谱系细胞的影响。采用竞争性HSC移植和二次移植来检测Dlk1 KO对HSC长期造血再增殖潜力的影响。 RNA-Seq 和细胞代谢测定用于确定潜在机制。相对于祖细胞和成熟谱系细胞，Dlk1 在成年小鼠长期 HSC (LT-HSC) 中高度表达。成年小鼠 HSC 中的 Dlk1 KO 促使 HSC 进入活跃的细胞周期，并扩大表型 LT-HSC，但破坏了其长期造血再增殖潜力。 Dlk1 KO导致HSCs代谢活性增加，包括葡萄糖摄取、核糖体翻译、线粒体代谢和ROS产生，从而损害HSCs功能。此外，成年小鼠 HSC 中的 Dlk1 KO 减弱了 Notch 信号传导，而 Dlk1 KO 下 Notch 信号传导的重新激活降低了线粒体活性和 ROS 产生，并挽救了 HSC 频率和绝对数量的变化。通过抗氧化剂N-乙酰半胱氨酸清除ROS可以抑制线粒体代谢活性，并挽救Dlk1 KO引起的HSC变化。我们的研究表明，Dlk1 在维持 HSC 稳态中发挥着重要作用，这是通过控制细胞周期和限制线粒体代谢活动来实现的。在线版本包含可在 10.1186/s40164-022-00369-9 获取的补充材料。
Adult hematopoietic stem cells (HSCs) homeostasis is critically important in maintaining lifelong hematopoiesis. However, how adult HSCs orchestrate its homeostasis remains not fully understood. Imprinted gene Dlk1 has been shown to play critical role in mouse embryonic hematopoiesis and in regulation of stem cells, but its physiological roles in adult HSCs are unknown. We performed gene expression analysis of Dlk1, and constructed conditional Dlk1 knockout (KO) mice by crossing Mx1 cre mice with Dlkflox/flox mice. Western blot and quantitative PCR were used to detect Dlk1 KO efficiency. Flow cytometry was performed to investigate the effects of Dlk1 KO on HSCs, progenitors and linage cells in primary mice. Competitive HSCs transplantation and secondary transplantation was used to examine the effects of Dlk1 KO on long-term hematopoietic repopulation potential of HSCs. RNA-Seq and cell metabolism assays was used to determine the underlying mechanisms. Dlk1 was highly expressed in adult mice long-term HSCs (LT-HSCs) relative to progenitors and mature lineage cells. Dlk1 KO in adult mice HSCs drove HSCs enter active cell cycle, and expanded phenotypical LT-HSCs, but undermined its long-term hematopoietic repopulation potential. Dlk1 KO resulted in an increase in HSCs’ metabolic activity, including glucose uptake, ribosomal translation, mitochondrial metabolism and ROS production, which impaired HSCs function. Further, Dlk1 KO in adult mice HSCs attenuated Notch signaling, and re-activation of Notch signaling under Dlk1 KO decreased the mitochondrial activity and ROS production, and rescued the changes in frequency and absolute number of HSCs. Scavenging ROS by antioxidant N-acetylcysteine could inhibit mitochondrial metabolic activity, and rescue the changes in HSCs caused by Dlk1 KO. Our study showed that Dlk1 played an essential role in maintaining HSC homeostasis, which is realized by governing cell cycle and restricting mitochondrial metabolic activity. The online version contains supplementary material available at 10.1186/s40164-022-00369-9.
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影响因子：
2.6
作者：
Chou, Song;Flygare, Johan;Lodish, Harvey F.
通讯作者：
Lodish, Harvey F.
影响因子：
30.8
作者：
Cleaton MA;Dent CL;Howard M;Corish JA;Gutteridge I;Sovio U;Gaccioli F;Takahashi N;Bauer SR;Charnock-Jones DS;Powell TL;Smith GC;Ferguson-Smith AC;Charalambous M
通讯作者：
Charalambous M
影响因子：
--
作者：
Bray, Sarah J.;Takada, Shuji;Harrison, Emma;Shen, Shing-Chuan;Ferguson-Smith, Anne C.
通讯作者：
Ferguson-Smith, Anne C.
影响因子：
23.9
作者：
Lv K;Gong C;Antony C;Han X;Ren JG;Donaghy R;Cheng Y;Pellegrino S;Warren AJ;Paralkar VR;Tong W
通讯作者：
Tong W
影响因子：
4
作者：
Doggett, YL;Briggs, JA;Briggs, RC
通讯作者：
Briggs, RC