AUTHOR=Fu Yuting , Liu Fangyuan , Cao Shuo , Zhang Jia , Wang Huizhi , Wu Baojiang , Song Yongli , Duo Shuguang , Li Xihe , Bao Siqin 

TITLE=Bdh2 Deficiency Promotes Endoderm-Biased Early Differentiation of Mouse Embryonic Stem Cells

JOURNAL=Frontiers in Cell and Developmental Biology

VOLUME=9

YEAR=2021

URL=https://www.frontiersin.org/journals/cell-and-developmental-biology/articles/10.3389/fcell.2021.655145

DOI=10.3389/fcell.2021.655145

ISSN=2296-634X

ABSTRACT=<p>3-hydroxybutyrate dehydrogenase-2 (<italic>Bdh2</italic>), a short-chain dehydrogenase, catalyzes a rate-limiting step in the biogenesis of the mammalian siderophore, playing a key role in iron homeostasis, energy metabolism and apoptosis. However, the function of <italic>Bdh2</italic> in embryonic stem cells (ESCs) remains unknown. To gain insights into the role of <italic>Bdh2</italic> on pluripotency and cell fate decisions of mouse ESCs, we generated <italic>Bdh2</italic> homozygous knockout lines for both mouse advanced embryonic stem cell (ASC) and ESC using CRISPR/Cas9 genome editing technology. <italic>Bdh2</italic> deficiency in both ASCs and ESCs had no effect on expression of core pluripotent transcription factors and alkaline phosphatase activity, suggesting dispensability of <italic>Bdh2</italic> for self-renewal and pluripotency of ESCs. Interestingly, cells with <italic>Bdh2</italic> deficiency exhibited potency of endoderm differentiation <italic>in vitro</italic>; with upregulated endoderm associated genes revealed by RNA-seq and RT-qPCR. We further demonstrate that <italic>Bdh2</italic> loss inhibited expression of multiple methyltransferases (DNMTs) at both RNA and protein level, suggesting that <italic>Bdh2</italic> may be essentially required to maintain DNA methylation in ASCs and ESCs. Overall, this study provides valuable data and resources for understanding how <italic>Bdh2</italic> regulate earliest cell fate decision and DNA methylation in ASCs/ESCs.</p>