Insights Into The Cell Fate Decision Of The Leydig-like Cells Reprogrammed From Mouse Fibroblasts

Objective:To investigate the role of transcription factors combination or small molecule compounds in determining the fate of mouse embryonic fibroblasts reprogrammed into Leydig cells and its possible molecular mechanisms.Our study provides a theoretical basis and method for induced Leydig cells with more basic research and clinical application value in the future.Methods:We downloaded the RNA-Seq data sets(GSE87020,GSE145797)respectively from the National Center for Biotechnology Information,screened and homogenized the raw data before performing similarity analysis of gene expression profile between samples in the transcription factors combination induction group and small molecule compounds induction group.After preliminary screening of differentially expressed genes,GO annotation and pathway enrichment were applicated.In addition,pathway enrichment analysis was performed on the expression levels of all transcription factors in both groups.Result:Transcriptome data showed that the genes expressed in embryonic fibroblasts of different types of mice are different.Compared with C57BL/7 mice,Kunming mice fibroblasts expresses more extracellular matrix,while the gene expression levels of Leydig cell differentiation and gonadal development are lower.Both combination of transcription factors and small molecule compounds can induce fibroblasts to undergo trans-differentiation.However,at the level of trans-differentiation,transcription factors combination is more effective.Several key genes,including Mrpl36,Rchy1,Atp5 b,N4bp1,Lcn2,Adh1,Mrap are expressed at levels comparable to those found in wild-type LCs.Wikipath analysis showed that 58 transcription factors including Cox2 are activated,and these factors are also concentrated in the cholesterol pathway,lipid metabolism,insulin Pathway,etc.It indicated that the combination of transcription factors can directly activate the development and differentiation of Leydig cells and the function of androgen synthesis.The combination of small molecule compounds can also activate a variety of transcription factors,including the MEF2/NR4A1 pathway,which are closely related to the expression of key enzymes in the androgen synthesis pathway of the Leydig cells.Conclusion:Compared with wild-type LC,the expression profiles of Ti LC and Ci LC are closer to MEF.Ci LC has more differentially expressed genes than Ti LC,while GO functional annotation and pathway enrichment showed that the combination of transcription factors prefers to promote the transformation of MEF into Leydig-like cells than chemicals induction.Compared with Ci LC,Ti LC has more transcription factors with a similar expression level as wild-type LC,and the expression trend is unidirectional,but Ci LC tend to bidirectional.Therefore,in the process of embryonic fibroblasts transdifferentiating into Leydig cells,the choice of transcription factors combination is superior to the combination of small molecule compounds,while in the expression of transcription factors and key enzymes in androgen synthesis,MEF <Ci LC <Ti LC < LC.