RUNX1-205, a novel splice variant of the human RUNX1 gene, has blockage effect on mesoderm-hemogenesis transition and promotion effect during the late stage of hematopoiesis.

Bo Chen,Feng Ma,Guohui Bian,Jiahui Zeng,Jiaxing Liu,Jing Chang,Mowen Lai,Qiongxiu Zhou,Wencui Sun,Xu Pan,Ya Zhou,Yonggang Zhang,Yuan Xue

doi:10.1093/jmcb/mjaa019

Abstract

Runt-related transcription factor 1 (RUNX1) is required for definitive hematopoiesis; however, the functions of most human RUNX1 isoforms are unclear. In particular, the effects of RUNX1-205 (a novel splice variant that lacks exon 6 in comparison with RUNX1b) on human hematopoiesis are not clear. In this study, a human embryonic stem cell (hESC) line with inducible RUNX1-205 overexpression was established. Analyses of these cells revealed that induction of RUNX1-205 overexpression at early stage did not influence the induction of mesoderm but blocked the emergence of CD34+ cells, and the production of hematopoietic stem/progenitor cells was significantly reduced. In addition, the expression of hematopoiesis-related factors was downregulated. However, these effects were abolished when RUNX1-205 overexpression was induced after Day 6 in co-cultures of hESCs and AGM-S3 cells, indicating that the inhibitory effect occurred prior to generation of hemogenic endothelial cells, while the promotive effect could be observed during the late stage of hematopoiesis. This is very similar to that of RUNX1b. Interestingly, the mRNA expression profile of RUNX1-205 during hematopoiesis was distinct from that of RUNX1b, and the protein stability of RUNX1-205 was much higher than that of RUNX1b. Thus, the function of RUNX1-205 in normal and diseased models should be further explored.

Highlights

Runt-related transcription factor 1 (RUNX1) is the key gene for human hematopoiesis, which plays a critical role in the development of hemogenic endothelium and hematopoietic stem cell (HSC) formation during embryogenesis (Chen et al, 2009)
A BLAST search of higher vertebrates from fish to human revealed that splice variants homologous to RUNX1-205 have been highly conserved during evolution (Supplementary Figure S1C)
In comparison with the noninduced control, induction of RUNX1-205 overexpression from Day 0 (D0) significantly blocked formation of burst-forming unit-erythroid (BFU-E), colony-forming unit-erythroid (CFU-E), colony-forming unit-mixed (CFU-Mix), and colony-forming unit-granulocyte/macrophage (CFU-GM) colonies (P < 0.05); the colony numbers were normal with RUNX1-205 induction from D6 (Figure 4A), consistent with the Fluorescence activated cell sorting (FACS) results (Figure 2B and C; Supplementary Figure S3B and C). These results indicate that overexpression of RUNX1-205 at early stage could block human hematopoiesis, similar to RUNX1b

Summary

Introduction

Runt-related transcription factor 1 (RUNX1) is the key gene for human hematopoiesis, which plays a critical role in the development of hemogenic endothelium and hematopoietic stem cell (HSC) formation during embryogenesis (Chen et al, 2009). Embryonic stem cells of Runx knockout mouse are unable to undergo hematopoietic differentiation, which can be rescued by Runx re-expression (Nishimura et al, 2004).

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Results

Conclusion