Probing the signaling requirements for naive human pluripotency by high-throughput chemical screening

• DOI: 10.1016/j.celrep.2021.109233
• Publication Date: 2021-06-15T14:45:06Z
• Authors: Shafqat A. Khan; Kyoung-mi Park; Laura A. Fischer; Chen Dong; Tenzin Lungjangwa; Marta Jimenez; Dominick Casalena; Brian Chew; Sabine Dietmann; Douglas S. Auld; Rudolf Jaenisch; Thorold W. Theuni...
• Keywords: Epiblast; Nodal signaling

Naive human embryonic stem cells (hESCs) have been isolated that more closely resemble the pre-implantation epiblast compared to conventional "primed" hESCs, but signaling principles underlying these discrete cell states remain incompletely understood. Here, we describe results from a high-throughput screen using ∼3,000 well-annotated compounds identify essential requirements for naive pluripotency. We report MEK1/2 inhibitors can be replaced during maintenance of pluripotency by targeting either upstream (FGFR, RAF) or downstream (ERK1/2) kinases. hESCs maintained under alternative conditions display elevated levels ERK phosphorylation retain genome-wide DNA hypomethylation and transcriptional identity epiblast. In contrast, dual inhibition MEK promotes efficient primed-to-naive resetting in combination with PKC, ROCK, TNKS activin A. This work demonstrates induction are governed distinct requirements.