MYC Controls Human Pluripotent Stem Cell Fate Decisions through Regulation of Metabolic Flux

Abstract

SUMMARY As human pluripotent stem cells (hPSCs) exit pluripotency they are thought to switch from a glycolytic mode of energy generation to one more dependent on oxidative phosphorylation. Here we show that although metabolic switching occurs during early mesoderm and endoderm differentiation, high glycolytic flux is maintained and in fact essential during early ectoderm specification. The elevated glycolysis observed in hPSCs requires elevated MYC/MYCN activity. Metabolic switching during endodermal and mesodermal differentiations coincides with a reduction in MYC/MYCN, and can be reversed by ectopically restoring MYC activity. During early ectodermal differentiation, sustained MYCN activity maintains the transcription of 'switch' genes that are rate-limiting for metabolic activity and lineage commitment. Our work, therefore, shows that metabolic switching is lineage-specific and not a required step for exit of pluripotency in hPSCs, and identifies MYC and MYCN as developmental regulators that couple metabolism to pluripotency and cell fate determination.

Revisiting the role of metabolism during development

“…Remarkably, the remodeling of energy metabolism during somatic cell reprogramming precedes the upregulation of pluripotency marker genes (Folmes et al, 2011;Kida et al, 2015). Moreover, lineagespecific metabolic rewiring is essential for commitment to a specific lineage (Cliff et al, 2017;Zheng et al, 2016). These examples suggest the possibility that cellular energy metabolism plays an instructive role in cell fate decisions.…”

Endogenous pluripotent factor expression after reprogramming cat fetal fibroblasts using inducible transcription factors

“…While all five transgenes remained expressed at P4, exogenous NANOG, SOX3 , and POU5FI were silenced by P14. The KLF4 and cMYC transgenes, however, remained active throughout 36 passages, thus limiting the usefulness of the iPSC as these genes are linked with differentiation and tumorigenesis (Cliff et al, ; Hu et al, ; Lin et al, ; Nakagawa et al, ; Ohnuki et al, ). Therefore, the complete transgene‐silencing achieved using the inducible system overcomes this limitation previously observed in cat and domestic species (Soto & Ross, ).…”

“…In our study, a lentiviral-based, drug inducible system was used to generate potential iPSCs in the domestic cat. passages, thus limiting the usefulness of the iPSC as these genes are linked with differentiation and tumorigenesis (Cliff et al, 2017;Hu et al, 2015;Lin et al, 2012;Nakagawa et al, 2008;Ohnuki et al, 2014). Therefore, the complete transgene-silencing achieved using the inducible system overcomes this limitation previously observed in cat and domestic species (Soto & Ross, 2016).…”

Human Pluripotent Stem Cell-Derived Multipotent Vascular Progenitors of the Mesothelium Lineage Have Utility in Tissue Engineering and Repair

“…Maintenance of pluripotent cells (Cliff et al, 2017) was as previously described. Briefly, WA09 (Sex: female, WiCell, NIHhESC-10-0062), WA07 (Sex: male, WiCell, NIHhESC-10-0061), WA01 (Sex: male, WiCell, NIHhESC-10-0043), TE03 (Sex: female, WiCell, NIHhESC-13-0204) human embryonic stem cells (hESCs) and K3 human induced pluripotent stem cells (hiPSCs), a gift from Stephen Duncan (Si-Tayebet al, 2010), were maintained in chemically-defined media (CDM) (Wang et al, 2007), seeded at 5 × 10 4 cells/cm 2 on Geltrex (Thermo Fisher, A1413302) coated plates at 1:200 dilution in DMEM/F12 w/o glutamine (Corning, 15-090-CM), and passaged with Accutase (Innovative Cell Technologies, AT104) upon confluency.…”