Metabolic regulation of pluripotency and germ cell fate through α-ketoglutarate.

John Reid,Lorenz Wernisch,M Azim Surani,Edward Allgeyer,Ben D Simons,Wolfram H Gruhn,Julia Tischler,Carsten Marr,Fabian Theis,Florian Buettner

doi:10.15252/embj.201899518

Abstract

An intricate link is becoming apparent between metabolism and cellular identities. Here, we explore the basis for such a link in an in vitro model for early mouse embryonic development: from naïve pluripotency to the specification of primordial germ cells (PGCs). Using single‐cell RNA‐seq with statistical modelling and modulation of energy metabolism, we demonstrate a functional role for oxidative mitochondrial metabolism in naïve pluripotency. We link mitochondrial tricarboxylic acid cycle activity to IDH2‐mediated production of alpha‐ketoglutarate and through it, the activity of key epigenetic regulators. Accordingly, this metabolite has a role in the maintenance of naïve pluripotency as well as in PGC differentiation, likely through preserving a particular histone methylation status underlying the transient state of developmental competence for the PGC fate. We reveal a link between energy metabolism and epigenetic control of cell state transitions during a developmental trajectory towards germ cell specification, and establish a paradigm for stabilizing fleeting cellular states through metabolic modulation.

Highlights

Embryonic stem cells (ESCs) have the capacity for indefinite selfrenewal in vitro, while retaining the ability to differentiate into specialized cell types (Ng & Surani, 2011; Young, 2011)
Aerobic glycolysis has been linked to chromatin structure and the maintenance of human ESCs (hESCs) pluripotency, with glycolysis-derived cytosolic acetyl-CoA serving as an essential substrate for histone acetylation (Moussaieff et al, 2015)
We harnessed the cellular heterogeneity arising during epiblast-like cells (EpiLCs) differentiation to derive dynamic gene expression trajectories by statistically ordering single-cell transcriptomes over a developmental time (“pseudotime”; Trapnell et al, 2014; Reid & Wernisch, 2016; Figs 1B and EV1D), and comprehensively quantified expression level changes (Appendix Table S1)

Summary

Introduction

Embryonic stem cells (ESCs) have the capacity for indefinite selfrenewal in vitro, while retaining the ability to differentiate into specialized cell types (Ng & Surani, 2011; Young, 2011). The in vitro differentiation of mouse ESCs (mESCs) from a naıve pluripotent state into primed epiblast-like cells (EpiLCs) confers transient developmental competence for the primordial germ cell (PGC) fate (Hayashi et al, 2011) and provides a tractable model system for investigations on early embryonic cell state conversions (Fig 1A). These cell states and their transitions are associated with functional heterogeneity, which needs consideration (Cahan & Daley, 2013). The precise molecular regulation underlying the impact of energy metabolism on mESC pluripotency and during early embryonic development, remains poorly defined

Methods

Results

Conclusion