ZNF281/Zfp281 is a target of miR-1 and counteracts muscle differentiation.

Sara Nicolai,Artem Smirnov,Giuseppe Raschellà,Eleonora Candi,Marco Pieraccioli,Alessandro Mauriello,Ying Wang,Margherita Annicchiarico‐Petruzzelli,Consuelo Pitolli,Gerry Melino,Lucia Anemona,Yufang Shi

doi:10.1002/1878-0261.12605

Abstract

Defects in achieving a fully differentiated state and aberrant expression of genes and microRNAs (miRs) involved in differentiation are common to virtually all tumor types. Here, we demonstrate that the zinc finger transcription factor ZNF281/Zfp281 is down‐regulated during epithelial, muscle, and granulocytic differentiation in vitro. The expression of this gene is absent in terminally differentiated human tissues, in contrast to the elevated expression in proliferating/differentiating ones. Analysis of the 3’UTR of ZNF281/Zfp281 revealed the presence of numerous previously undescribed miR binding sites that were proved to be functional for miR‐mediated post‐transcriptional regulation. Many of these miRs are involved in differentiation pathways of distinct cell lineages. Of interest, ZNF281/Zfp281 is able to inhibit muscle differentiation promoted by miR‐1, of which ZNF281/Zfp281 is a direct target. These data suggest that down‐regulation of ZNF281/Zfp281 during differentiation in various cell types may occur through specific miRs whose expression is tissue‐restricted. In addition, we found that in rhabdomyosarcoma and leiomyosarcoma tumors, the expression of ZNF281/Zfp281 is significantly higher compared with normal counterparts. We extended our analysis to other human soft tissue sarcomas, in which the expression of ZNF281 is associated with a worse prognosis. In summary, we highlight here a new role of ZNF281/Zfp281 in counteracting muscle differentiation; its down‐regulation is at least in part mediated by miR‐1. The elevated expression of ZNF281/Zfp281 in soft tissue sarcomas warrants further analysis for its possible exploitation as a prognostic marker in this class of tumors.

Highlights

A variable degree of dedifferentiation compared with the normal tissue they originate from is a distinctive feature of tumors (Merrell and Stanger, 2016) and other pathologies (Xie et al, 2018)
Since ZNF281/Zfp281 has been detected in many normal tissues during embryonic and adult life (Fidalgo et al, 2011; Pieraccioli et al, 2018; Zhou et al, 2017), we looked for a flexible regulatory mechanism that can effectively regulate its expression in different cellular contexts
The amount of Zfp281 protein did not significantly change in miR-23a/b-transfected cells compared with controls (Fig. S1C), confirming that miR23a/b are unable to affect the expression of Zfp281 in mice

Summary

Introduction

A variable degree of dedifferentiation compared with the normal tissue they originate from is a distinctive feature of tumors (Merrell and Stanger, 2016) and other pathologies (Xie et al, 2018). The zinc finger transcription factor ZNF281/Zfp281 (ZNF281 in humans, Zfp281 in mice) acts as a stemness regulator mediating Nanog autorepression during embryonic/fetal life (Fidalgo et al, 2011). The biological activity of ZNF281/Zfp281 is not limited to embryonic/fetal life; Zfp281 increases cardiac reprogramming of adult mouse fibroblasts by modulating the expression of inflammatory and cardiac genes (Zhou et al, 2017); in normal and transformed cells, ZNF281/Zfp281 down-regulation promotes osteogenic (Seo et al, 2013) and neuronal differentiation (Pieraccioli et al, 2018), while its expression drives epithelial–mesenchymal transition (EMT) in colon cancer cells (Hahn et al, 2013). A further and somehow unexpected finding highlighted the involvement of ZNF281 in the transcriptional control of DNA damage repair genes (Pieraccioli et al, 2016), as well as in the recruitment of the repair machinery components directly on damage sites (Nicolai et al, 2019)

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Discussion

Conclusion