Maternal Undernutrition Induces Cell Signalling and Metabolic Dysfunction in Undifferentiated Mouse Embryonic Stem Cells

Abstract

Peri-conceptional environment can induce permanent changes in embryo phenotype which alter development and associate with later disease susceptibility. Thus, mouse maternal low protein diet (LPD) fed exclusively during preimplantation is sufficient to lead to cardiovascular, metabolic and neurological dysfunction in adult offspring. Embryonic stem cell (ESC) lines were generated from LPD and control NPD C57BL/6 blastocysts and characterised by transcriptomics, metabolomics, bioinformatics and molecular/cellular studies to assess early potential mechanisms in dietary environmental programming. Previously, we showed these lines retain cellular and epigenetic characteristics of LPD and NPD embryos after several passages. Here, three main changes were identified in LPD ESC lines. First, their derivation capacity was reduced but pluripotency marker expression was similar to controls. Second, LPD lines had impaired Mitogen-activated protein kinase (MAPK) pathway with altered gene expression of several regulators (e.g., Maff, Rassf1, JunD), reduced ERK1/2 signalling capacity and poorer cell survival characteristics which may contribute to reduced derivation. Third, LPD lines had impaired glucose metabolism comprising reduced upstream enzyme expression (e.g., Gpi, Mpi) and accumulation of metabolites (e.g., glucose-6-P, fructose-6-P) above the phosphofructokinase (PFK) gateway with PFK enzyme activity reduced. ESC lines may therefore permit investigation of peri-conceptional programming mechanisms with reduced need for animal experimentation. Graphical Abstract

Unraveling the Molecular Mechanisms of the Neurodevelopmental Consequences of Fetal Protein Deficiency: Insights From Rodent Models and Public Health Implications

“…This remarkably short period of protein restriction reduced BCAA levels in blastocysts and blocked glucose metabolism, the primary energy source at this stage. In embryonic stem cells cultured from G3.5 blastocysts, MAPK (mitogen-activated protein kinase), ERK1/2 (extracellular signal-regulated kinase), and mTORC1 (mechanistic target of rapamycin complex 1) signaling pathways were dysregulated ( 92 , 93 ). Cell-tracing and immunohistochemical studies up to G12 showed reduced progenitor proliferation coupled with increased neuronal differentiation in the basal ganglia and cortex.…”

The mammalian preimplantation embryo: Its role in the environmental programming of postnatal health and performance

“…The latter is considered a compensatory mechanism to increase histotrophic nutrition during episodes of protein deprivation and can be induced in vitro by deficient BCAA availability, especially isoleucine (Caetano et al, 2021). There is also evidence of deficiencies in MAP kinase signalling associated with increased apoptosis, and in glycolysis affecting energy metabolism in embryonic stem cell lines derived from maternal protein restricted ICMs (Khurana et al, 2023).…”