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Zebrafish miR-462-731 regulates hematopoietic specification and pu.1-dependent primitive myelopoiesis

Cell Death & Differentiation (2018) | Download Citation

Abstract

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.1-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.

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Edited by: S. Kumar.

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Acknowledgements

We are grateful to Prof. Anming Meng, Tsinghua University, for providing the Tg(flk1:GFP/gata1:DsRed) transgenic line. This work was supported by Natural Science Foundation of China (31872542), Top-notch Talent Support Program of College of Fishery, the Scientific Research Foundation for the Returned Overseas Chinese Scholars, State Education Ministry, and the Fundamental Research Funds for the Central Universities (2016PY030).

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  1. These authors contributed equally: Chun-Xiao Huang, Yan Huang.

Affiliations

  1. Key Lab of Freshwater Animal Breeding, Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction, Ministry of Education, College of Fishery, Huazhong Agricultural University, Wuhan, Hubei, P.R. China

    • Chun-Xiao Huang
    • , Yan Huang
    • , Xue-Ke Duan
    • , Mu Zhang
    • , Jia-Peng Tu
    • , Jing-Xia Liu
    • , Hong Liu
    • , Tian-Sheng Chen
    • , Wei-Min Wang
    •  & Huan-Ling Wang

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The authors declare that they have no conflict of interest.

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Correspondence to Huan-Ling Wang.

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https://doi.org/10.1038/s41418-018-0234-0