SF3B1 deficiency impairs human erythropoiesis via activation of p53 pathway: implications for understanding of ineffective erythropoiesis in MDS.
SF3B1 deficiency impairs human erythropoiesis via activation of p53 pathway: implications for understanding of ineffective erythropoiesis in MDS.
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SF3B1 缺陷通过激活 p53 通路损害人类红细胞生成：对理解 MDS 无效红细胞生成的影响
DOI：
10.1186/s13045-018-0558-8
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发表时间：
2018-02-12
影响因子：
28.5
通讯作者：
An X
中科院分区：
文献类型：
--
作者：
Huang Y;Hale J;Wang Y;Li W;Zhang S;Zhang J;Zhao H;Guo X;Liu J;Yan H;Yazdanbakhsh K;Huang G;Hillyer CD;Mohandas N;Chen L;Sun L;An X
dsf3b1是拼接机械的核心部件。SF3B1突变经常在骨髓增生异常综合征（MDS）中发现，特别是在具有环状铁母细胞（RARS）的难治性贫血患者中，其特征是孤立性贫血。SF3B1突变与RARS的病理生理有关；然而，SF3B1在红细胞生成中的生理功能尚不清楚。方法采用shrna介导的方法敲除人CD34+细胞中的SF3B1。流式细胞术观察SF3B1基因敲低对人红细胞分化、细胞周期和凋亡的影响。采用RNA-seq、qRT-PCR和western blot分析来确定SF3B1基因敲低后表型的机制。结果在人CD34+细胞中，SF3B1敲低可导致早期红细胞凋亡增加和细胞周期阻滞，并产生异常成核的晚期红母细胞。sf3b1敲低的红系祖细胞CFU-E的RNA-seq分析显示E3连接酶Makorin Ring Finger Protein 1 （MKRN1）剪接改变，随后p53通路激活。重要的是，MKRN1的异位表达挽救了sf3b1敲低诱导的改变。与对照细胞相比，sf3b1基因敲除的多色和正色红细胞中有丝分裂/细胞分裂途径相关基因的表达减少，包括polo样激酶1 （PLK1）。PLK1的药理学抑制也导致异常成核红细胞的产生。结论这些发现使我们能够确定SF3B1在人红细胞生成中的新作用，并为其在正常红细胞生成中的调节作用提供了新的见解。此外，这些发现有助于更好地理解SF3B1突变MDS患者的无效红细胞生成。
BackgroundSF3B1 is a core component of splicing machinery. Mutations in SF3B1 are frequently found in myelodysplastic syndromes (MDS), particularly in patients with refractory anemia with ringed sideroblasts (RARS), characterized by isolated anemia. SF3B1 mutations have been implicated in the pathophysiology of RARS; however, the physiological function of SF3B1 in erythropoiesis remains unknown.MethodsshRNA-mediated approach was used to knockdown SF3B1 in human CD34+cells. The effects of SF3B1 knockdown on human erythroid cell differentiation, cell cycle, and apoptosis were assessed by flow cytometry. RNA-seq, qRT-PCR, and western blot analyses were used to define the mechanisms of phenotypes following knockdown of SF3B1.ResultsWe document that SF3B1 knockdown in human CD34+cells leads to increased apoptosis and cell cycle arrest of early-stage erythroid cells and generation of abnormally nucleated late-stage erythroblasts. RNA-seq analysis of SF3B1-knockdown erythroid progenitor CFU-E cells revealed altered splicing of an E3 ligase Makorin Ring Finger Protein 1 (MKRN1) and subsequent activation of p53 pathway. Importantly, ectopic expression of MKRN1 rescued SF3B1-knockdown-induced alterations. Decreased expression of genes involved in mitosis/cytokinesis pathway including polo-like kinase 1 (PLK1) was noted in SF3B1-knockdown polychromatic and orthochromatic erythroblasts comparing to control cells. Pharmacologic inhibition of PLK1 also led to generation of abnormally nucleated erythroblasts.ConclusionsThese findings enabled us to identify novel roles for SF3B1 in human erythropoiesis and provided new insights into its role in regulating normal erythropoiesis. Furthermore, these findings have implications for improved understanding of ineffective erythropoiesis in MDS patients with SF3B1 mutations.
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影响因子：
11.4
作者：
通讯作者：
--
DOI：
10.1182/asheducation-2009.1.87
发表时间：
2009
期刊：
Hematology. American Society of Hematology. Education Program
影响因子：
--
作者：
Haldar K;Mohandas N
通讯作者：
Mohandas N
影响因子：
15.8
作者：
Ebert, Benjamin L.;Galili, Naomi;Raza, Azra
通讯作者：
Raza, Azra
影响因子：
82.9
作者：
Dussiot M;Maciel TT;Fricot A;Chartier C;Negre O;Veiga J;Grapton D;Paubelle E;Payen E;Beuzard Y;Leboulch P;Ribeil JA;Arlet JB;Coté F;Courtois G;Ginzburg YZ;Daniel TO;Chopra R;Sung V;Hermine O;Moura IC
通讯作者：
Moura IC
影响因子：
8
作者：
Cantor, AB;Orkin, SH
通讯作者：
Orkin, SH