Mechanisms Of Forkhead Family Transcription Factors In Mouse Somatic Reprogramming

Somatic cell reprogramming is an ideal model to explore the mechanism of cell fate regulation.Transcription factors play a key regulatory role in cell fate transition.Studying the new molecular functions of transcription factors on the basis of somatic cell reprogramming models is of great significance for expanding the mechanism of transcription factors in determining cell fate,establishing a new reprogramming technology system,and obtaining function lineage cells.Forkhead family of transcription factors play an important role in embryonic development,disease occurrence and cell lineage transdifferentiation,but there are few studies for Forkhead family transcription factors on regulation of pluripotency,especially in the process of somatic cell reprogramming in mice.In this study,we firstly collected the high-throughput sequencing data such as ATAC-seq and single-cell RNA-seq of the mouse somatic cell reprogramming system and early embryonic development published in our laboratory and others.By using bioinformatics analysis,We depicted the dynamic changes of chromatin opening and closing in different systems of mouse somatic reprogramming,the distribution of different reprogramming branches,and the expression changes of different lineage cells during mouse early embryonic development.Among them,the data analysis results suggest that the FoxD and Fox R subgroups may be related to the regulation of pluripotency,and Fox J3,Fox R1 and FoxO subgroups may play an important role in early embryonic development.In order to further verify these results and explore the relationship between Forkhead family transcription factors and pluripotency regulation network,we cloned all members of mouse Forkhead family transcription factors.In the Oct4,Klf4 and Sox2 reprogramming system based on i CD1 medium,the effect of Forkhead family transcription factors on the generation of i PSCs was systematically explored.It was found that FoxD subfamily and FoxG1 could accelerate the generation of i PSCs on day 4.More importantly,FoxD3,FoxD4 and FoxG1 could replace Oct4 to produce germline transmitted i PSCs with Sox2 and Klf4.In contrast,Fox A and FoxO subfamilies significantly inhibit somatic reprogramming,in which FoxO6 inhibits this process by inhibiting cell proliferation,suppressing the expression of pluripotent genes and hindering the process of mesenchymal to epithelial transition.Thus,this study sheds light on the role of Forkhead transcription factors in mouse somatic cell reprogramming,and provides new knowledge and insights to regulate cell fate transition and establish a new reprogramming technology system.