Disrupting Interactions Between beta-Catenin and Activating TCFs Reconstitutes Ground State Pluripotency in Mouse Embryonic Stem Cells

作者：Saj Abil; Chatterjee Sujash S; Zhu Bowen; Cukuroglu Engin; Gocha Tenzin; Zhang Xiaoqian; Goke Jonathan; DasGupta Ramanuj^*

来源：Stem Cells, 2017, 35(8): 1924-1933.

DOI：10.1002/stem.2647

• Mouse embryonic stem cell
• iCRT3
• beta-catenin
• Pluripotency
• 2i-media

The 2i-media, composed of two small molecule inhibitors (PD0325901 and CHIR99021) against MEK and GSK3-kinases, respectively, is known to establish naive ground state pluripotency in mouse embryonic stem cells (mESCs). These inhibitors block MEK-mediated differentiation, while driving beta-catenin dependent de-repression of pluripotency promoting targets. However, accumulating evidence suggest that beta-catenin's association with activating TCFs (TCF7 and TCF7L2) can induce expression of several lineage-specific prodifferentiation genes. We posited that CHIR-induced upregulation of beta-catenin levels could therefore compromise the stability of the naive ive state in long-term cultures. Here, we investigated whether replacing CHIR with iCRT3, a small molecule that abrogates beta-catenin-TCF interaction, can still retain ground state pluripotency in mESCs. Our data suggests that iCRT3+PD mediated coinhibition of MEK and beta-catenin/TCF-dependent transcriptional activity over multiple passages significantly reduces expression of differentiation markers, as compared to 2i. Furthermore, the ability to efficiently contribute toward chimera generation and germline transmission suggests that the inhibition of beta-catenin's TCF-dependent transcriptional activity, independent of its protein expression level, retains the naive ground state pluripotency in mESCs. Additionally, growth medium containing iCRT3+PD can provide an alternative to 2i as a stable culture method.

• 出版日期2017-8