Zebrafish miR-462-731 regulates hematopoietic specification and pu.1-dependent primitive myelopoiesis
Zebrafish miR-462-731 regulates hematopoietic specification and pu.1-dependent primitive myelopoiesis
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斑马鱼 miR-462-731 调节造血规范和 pu.1 依赖性原始骨髓生成
DOI：
10.1038/s41418-018-0234-0
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发表时间：
2019-08-01
影响因子：
12.4
通讯作者：
Wang, Huan-Ling
中科院分区：
文献类型：
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作者：
Huang, Chun-Xiao;Huang, Yan;Wang, Huan-Ling
microRNAs（miRNAs）在胚胎造血和血液系统恶性肿瘤中发挥重要作用。斑马鱼miR-462-731簇是人类miR-191-425的正向同源簇，调控细胞增殖和肿瘤发生。在我们之前的工作中，发现miR-462-731在早期胚胎发生过程中高度且普遍表达。在这项研究中，通过功能丧失分析（吗啉代敲除结合CRISRP/Cas9敲除）和mRNA谱分析，我们表明miR-462-731通过调节细胞存活而为正常胚胎发育所需。我们发现，miR-462/miR-731的缺失导致48 hpf时红系细胞数量的显著减少以及异位髓系细胞扩增，表明髓系-红系谱系分化的偏斜。从机制上讲，miR-462-731为pu.通过调节etsrp/scl信号转导与新的pu结合来依赖原始骨髓生成。1/miR-462-731反馈环。另一方面，miR-462/miR-731的吗啉代（MO）敲低导致24 hpf时后血岛的扩张，这是由BMP信号传导升高导致的轻度腹侧化表型。使用BMP I型受体抑制剂dorsomorphin和alk 8 MO的拯救实验表明，miR-462-731在BMP/Smad信号传导途径内的alk 8上游起作用，并作为新的内源性BMP拮抗剂起作用。此外，在miR-462/miR-731变体中观察到血管生成受损。动脉和静脉的特化也受到干扰，其特征在于efnb 2a和flt 4表达的不规则模式。我们的研究揭示了miR-462-731在BMP/Smad信号介导的中胚层祖细胞造血特化中的先前未被认识的作用，并证明了miR-462-731介导的驱动ALPM中原始骨髓生成的调节机制。我们还显示了miR-462-731在调节动脉-静脉特化和永久性造血干细胞（HSC）产生中的需求。本研究结果可能为进一步了解miRNA调控胚胎造血和血液系统恶性肿瘤的分子机制基础提供依据。
MicroRNAs (miRNAs) play significant roles in both embryonic hematopoiesis and hematological malignancy. Zebrafish miR-462-731 cluster is orthologous of miR-191-425 in human which regulates proliferation and tumorigenesis. In our previous work, miR-462-731 was found highly and ubiquitously expressed during early embryogenesis. In this study, by loss-of-function analysis (morpholino knockdown combined with CRISRP/Cas9 knockout) and mRNA profiling, we suggest that miR-462-731 is required for normal embryonic development by regulating cell survival. We found that loss of miR-462/miR-731 caused a remarkable decrease in the number of erythroid cells as well as an ectopic myeloid cell expansion at 48 hpf, suggesting a skewing of myeloid-erythroid lineage differentiation. Mechanistically, miR-462-731 provides an instructive input for pu./-dependent primitive myelopoiesis through regulating etsrp/scl signaling combined with a novel pu. 1/miR-462-731 feedback loop. On the other hand, morpholino (MO) knockdown of miR-462/miR-731 resulted in an expansion of posterior blood islands at 24 hpf, which is a mild ventralization phenotype resulted from elevation of BMP signaling. Rescue experiments with both BMP type I receptor inhibitor dorsomorphin and alk8 MO indicate that miR-462-731 acts upstream of alk8 within the BMP/Smad signaling pathway and functions as a novel endogenous BMP antagonist. Besides, an impairment of angiogenesis was observed in miR-462/miR-731 morphants. The specification of arteries and veins was also perturbed, as characterized by the irregular patterning of efnb2a and flt4 expression. Our study unveils a previously unrecognized role of miR-462-731 in BMP/Smad signaling mediated hematopoietic specification of mesodermal progenitors and demonstrates a miR-462-731 mediated regulatory mechanism driving primitive myelopoiesis in the ALPM. We also show a requirement for miR-462-731 in regulating arterial-venous specification and definitive hematopoietic stem cell (HSC) production. The current findings might provide further insights into the molecular mechanistic basis of miRNA regulation of embryonic hematopoiesis and hematological malignancy.