OVOL1 Influences the Determination and Expansion of iPSC Reprogramming Intermediates

Abstract

Summary: During somatic cell reprogramming to induced pluripotent stem cells (iPSCs), fibroblasts undergo dynamic molecular changes, including a mesenchymal-to-epithelial transition (MET) and gain of pluripotency; processes that are influenced by Yamanaka factor stoichiometry. For example, in early reprogramming, high KLF4 levels are correlated with the induction of functionally undefined, transiently expressed MET genes. Here, we identified the cell-surface protein TROP2 as a marker for cells with transient MET induction in the high-KLF4 condition. We observed the emergence of cells expressing the pluripotency marker SSEA-1+ mainly from within the TROP2+ fraction. Using TROP2 as a marker in CRISPR/Cas9-mediated candidate screening of MET genes, we identified the transcription factor OVOL1 as a potential regulator of an alternative epithelial cell fate characterized by the expression of non-iPSC MET genes and low cell proliferation. Our study sheds light on how reprogramming factor stoichiometry alters the spectrum of intermediate cell fates, ultimately influencing reprogramming outcomes. : Woltjen and colleagues identified the transcription factor OVOL1 as a potential regulator of transient MET induction in high-KLF4 reprogramming. Transient MET induction inhibits the expansion of intermediate partially reprogrammed cells. Their study provides insight into how reprogramming factor stoichiometry affects intermediate cell fates and reprogramming outcomes. Keywords: iPSC, reprogramming, stoichiometry, Klf4, mesenchymal-to-epithelial transition, Tacstd2, TROP2, Ovol1, SSEA-1, CRISPR/Cas9