Background and Aim: Heart diseases are the most significant cause of morbidity and mortality worldwide, which accounts for approximately ۳۳% of all deaths [۲]. Recently, a cellular alchemy-like method, known as
direct cardiac reprogramming (DCR) has been developed for direct conversion of somatic cells such as
fibroblast to cardiomyocytes required for cardiac cell-based therapy. To this end, several studies have reported TFs and miRNAs for DCR, but the efficiency of all DCR studies is low. Analysis of RNA-seq data of cardiac progenitors cells (CPCs) produced through DCR seems to be very useful for finding new TFs and miRNAs, which can be used along with previously reported factors to enhance the efficiency the DCR strategies, as well as the quality of produced CPCs.Methods: The RNA-seq data set GSE۷۷۳۷۵ was obtained from the GEO database that included fibroblasts and induced CPCs (iCPCs; generated via DCR strategy). Differential expressed genes (DEGs) were investigated using Galaxy (https://usegalaxy.org/) [with p values <۰.۰۵ and│logFC│≥۱.۵]. Gene Ontology (http://geneontology.org/) and KEGG (https://www.genome.jp/kegg/) were used to observe the biological processes and pathways associated with the genes upregulated in iCPCs. To find the TFs controlling the expression of upregulated genes, ChEA was used . We also predicted the miRNAs targeting the fibroblasts specific genes (downregulated genes) using enrichR. Finally, we constructed the protein-protein interaction network (PPI) using STRING (https://string-db.org/) to find most important factors, especially epigenetic regulating factors in iCPCs.Results: We found that ۲۴۷ and ۲۰۵۷ genes were upregulated and downregulated in iCPCs, respectively. The upregulated genes extensively involved in post-translational modification and extracellular matrix disassembly and organization. KEGG pathway analysis showed that p۵۳ and PI۳K-Akt signaling are the most significant pathways associated with upregulated genes in iCPCs. TFs analysis revealed that the top TFs associated with up DEGs were TNK۱, CSNK۲A۲ and MAPK۱۰ and in the other hand the top TFs for down DEGs were JAK۲, MAPK۱ and RET. We also found that mmu-miR-۲۷b-۳p and mmu-miR-۱۸۳-۵p are the most important miRNAs targeting downregulated genes in iCPCs. PPI network construction of iCPCs upregulated genes revealed that epigenetic regulating factors such as EZH۲, HDAC and SUZ۱۲ are in the core of PPI network.Conclusion: Activation of signaling pathway (PI۳K-Akt), epigenetic regulating mechanism (histone acetylation and methylation) along with overexpression of miRNAs (miR-۲۷b-۳p) might increase the quantity and the quality of iCPCs required for cardiac cell-based therapy.