Abstract
Pregnancy is a state of high metabolic demand. Fasting diverts metabolism to fatty acid oxidation, and the fasted response occurs much more rapidly in pregnant women than in non-pregnant women. The product of the imprinted DLK1 gene (delta-like homolog 1) is an endocrine signaling molecule that reaches a high concentration in the maternal circulation during late pregnancy. By using mouse models with deleted Dlk1, we show that the fetus is the source of maternal circulating DLK1. In the absence of fetally derived DLK1, the maternal fasting response is impaired. Furthermore, we found that maternal circulating DLK1 levels predict embryonic mass in mice and can differentiate healthy small-for-gestational-age (SGA) infants from pathologically small infants in a human cohort. Therefore, measurement of DLK1 concentration in maternal blood may be a valuable method for diagnosing human disorders associated with impaired DLK1 expression and to predict poor intrauterine growth and complications of pregnancy.
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References
Butte, N.F. Carbohydrate and lipid metabolism in pregnancy: normal compared with gestational diabetes mellitus. Am. J. Clin. Nutr. 71 (Suppl. 5), 1256S–1261S (2000).
Metzger, B.E., Ravnikar, V., Vileisis, R.A. & Freinkel, N. “Accelerated starvation” and the skipped breakfast in late normal pregnancy. Lancet 1, 588–592 (1982).
Schmidt, J.V., Matteson, P.G., Jones, B.K., Guan, X.J. & Tilghman, S.M. The Dlk1 and Gtl2 genes are linked and reciprocally imprinted. Genes Dev. 14, 1997–2002 (2000).
Takada, S. et al. Delta-like and Gtl2 are reciprocally expressed, differentially methylated linked imprinted genes on mouse chromosome 12. Curr. Biol. 10, 1135–1138 (2000).
Smas, C.M., Chen, L. & Sul, H.S. Cleavage of membrane-associated pref-1 generates a soluble inhibitor of adipocyte differentiation. Mol. Cell. Biol. 17, 977–988 (1997).
Bachmann, E., Krogh, T.N., Højrup, P., Skjødt, K. & Teisner, B. Mouse fetal antigen 1 (mFA1), the circulating gene product of mdlk, pref-1 and SCP-1: isolation, characterization and biology. J. Reprod. Fertil. 107, 279–285 (1996).
Floridon, C. et al. Does fetal antigen 1 (FA1) identify cells with regenerative, endocrine and neuroendocrine potentials? A study of FA1 in embryonic, fetal, and placental tissue and in maternal circulation. Differentiation 66, 49–59 (2000).
Carlsson, H.E., Persdotter-Hedlund, G., Fries, E., Eriksson, U.J. & Hau, J. Purification, characterization, and biological compartmentalization of rat fetal antigen 1. Biol. Reprod. 63, 30–33 (2000).
Smas, C.M. & Sul, H.S. Pref-1, a protein containing EGF-like repeats, inhibits adipocyte differentiation. Cell 73, 725–734 (1993).
Charalambous, M. et al. Imprinted gene dosage is critical for the transition to independent life. Cell Metab. 15, 209–221 (2012).
Charalambous, M. et al. DLK1/PREF1 regulates nutrient metabolism and protects from steatosis. Proc. Natl. Acad. Sci. USA 111, 16088–16093 (2014).
Moraitis, A.A., Wood, A.M., Fleming, M. & Smith, G.C. Birth weight percentile and the risk of term perinatal death. Obstet. Gynecol. 124, 274–283 (2014).
Raghunandan, R. et al. Dlk1 influences differentiation and function of B lymphocytes. Stem Cells Dev. 17, 495–507 (2008).
Chacón, M.R. et al. Human serum levels of fetal antigen 1 (FA1/Dlk1) increase with obesity, are negatively associated with insulin sensitivity and modulate inflammation in vitro. Int. J. Obes. (Lond.) 32, 1122–1129 (2008).
Rossant, J. & Cross, J.C. Placental development: lessons from mouse mutants. Nat. Rev. Genet. 2, 538–548 (2001).
Malik, N.M. et al. Leptin requirement for conception, implantation, and gestation in the mouse. Endocrinology 142, 5198–5202 (2001).
Schulz, L.C., Widmaier, E.P., Qiu, J. & Roberts, R.M. Effect of leptin on mouse trophoblast giant cells. Biol. Reprod. 80, 415–424 (2009).
Roman, E.A., Ricci, A.G. & Faletti, A.G. Leptin enhances ovulation and attenuates the effects produced by food restriction. Mol. Cell. Endocrinol. 242, 33–41 (2005).
da Rocha, S.T. et al. Gene dosage effects of the imprinted delta-like homologue 1 (Dlk1/Pref1) in development: implications for the evolution of imprinting. PLoS Genet. 5, e1000392 (2009).
Appelbe, O.K., Yevtodiyenko, A., Muniz-Talavera, H. & Schmidt, J.V. Conditional deletions refine the embryonic requirement for Dlk1. Mech. Dev. 130, 143–159 (2013).
Tallquist, M.D. & Soriano, P. Epiblast-restricted Cre expression in MORE mice: a tool to distinguish embryonic vs. extra-embryonic gene function. Genesis 26, 113–115 (2000).
Muzumdar, M.D., Tasic, B., Miyamichi, K., Li, L. & Luo, L. A global double-fluorescent Cre reporter mouse. Genesis 45, 593–605 (2007).
El-Kasti, M.M. et al. The pregnancy-induced increase in baseline circulating growth hormone in rats is not induced by ghrelin. J. Neuroendocrinol. 20, 309–322 (2008).
Soares, M.J. The prolactin and growth hormone families: pregnancy-specific hormones/cytokines at the maternal–fetal interface. Reprod. Biol. Endocrinol. 2, 51 (2004).
Pasupathy, D. et al. Study protocol. A prospective cohort study of unselected primiparous women: the Pregnancy Outcome Prediction Study. BMC Pregnancy Childbirth 8, 51 (2008).
Sovio, U., White, I.R., Dacey, A., Pasupathy, D. & Smith, G.C. Screening for fetal growth restriction with universal third trimester ultrasonography in nulliparous women in the Pregnancy Outcome Prediction (POP) study: a prospective cohort study. Lancet 386, 2089–2097 (2015).
Rotham, K.J.G. et al. Modern Epidemiology 3rd edn. (Lippincott-Raven, 2008).
de Zegher, F. et al. Abundance of circulating preadipocyte factor 1 in early life. Diabetes Care 35, 848–849 (2012).
Schrey, S. et al. The adipokine preadipocyte factor-1 is downregulated in preeclampsia and expressed in placenta. Cytokine 75, 338–343 (2015).
Moore, G.E. et al. The role and interaction of imprinted genes in human fetal growth. Phil. Trans. R. Soc. Lond. B 370, 20140074 (2015).
Kappil, M.A. et al. Placental expression profile of imprinted genes impacts birth weight. Epigenetics 10, 842–849 (2015).
Smith, G.C. Researching new methods of screening for adverse pregnancy outcome: lessons from pre-eclampsia. PLoS Med. 9, e1001274 (2012).
Ioannides, Y., Lokulo-Sodipe, K., Mackay, D.J., Davies, J.H. & Temple, I.K. Temple syndrome: improving the recognition of an underdiagnosed chromosome 14 imprinting disorder: an analysis of 51 published cases. J. Med. Genet. 51, 495–501 (2014).
Acknowledgements
M.A.M.C. was supported by a PhD studentship from the Cambridge Centre for Trophoblast Research. Research was supported by grants from the MRC (MR/J001597/1 and MR/L002345/1), the Medical College of Saint Bartholomew's Hospital Trust, a Wellcome Trust Investigator Award, EpigeneSys (FP7 Health-257082), EpiHealth (FP7 Health-278414), a Herchel Smith Fellowship (N.T.) and NIH grant RO1 DK89989. The contents are the authors' sole responsibility and do not necessarily represent official NIH views. We thank G. Burton for invaluable support, and M. Constância and I. Sandovici (University of Cambridge) for the Meox2-cre mice. We are extremely grateful to all of the participants in the Pregnancy Outcome Prediction study. This work was supported by the NIHR Cambridge Comprehensive Biomedical Research Centre (Women's Health theme) and project grants from the MRC (G1100221) and Sands (Stillbirth and Neonatal Death Charity). The study was also supported by GE Healthcare (donation of two Voluson i ultrasound systems for this study) and by the NIHR Cambridge Clinical Research Facility, where all research visits took place.
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M.C., M.A.M.C., A.C.F.-S. and G.C.S.S. conceived and designed the experiments. M.C., M.A.M.C., J.A.C., M.H., I.G., N.T., C.L.D. and D.S.C.-J. performed the experiments. M.C., M.A.M.C., A.C.F.-S., C.L.D., F.G. and G.C.S.S. analyzed the data. M.C. and U.S. performed statistical analysis. S.R.B., T.L.P., A.C.F.-S. and G.C.S.S. contributed reagents. M.C., M.A.M.C., A.C.F.-S. and G.C.S.S. wrote the manuscript. M.C., A.C.F.-S. and G.C.S.S. provided supervision.
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Integrated supplementary information
Supplementary Figure 1 Altered body composition in Null females can be detected during embryogenesis.
(a) Total bodyweight of virgin females at 12 weeks does not differ between genotypes. Null females have larger abdominal WAT deposits (b) and reduced muscle (combined gastrocnemius/soleus) (c) than WT and Mat females at 12 weeks. WT and Null n = 8, Mat n = 7, compared by One-way ANOVA, with Bonferroni's Multiple comparison post-hoc test with WT vs Null and WT vs Mat, *p<0.05. Body weight (d), crown rump (C-R) length (e) and tissue weights (f) in embryos at E18.5. Null embryos have reduced mass and length, and reduced lean (hindlimb and forelimb) mass than embryos expressing Dlk1, WT n > 21, Null n > 13, Mat n > 10, compared by Kruskall-Wallace test with Dunn's Multiple comparison post-hoc test comparing WT vs Null and WT vs Mat, *p<0.05, ***p<0.001. (g) Null mice are born small but catch up in the preweaning period. Serial measurements of pup weight from birth to weaning, Nulls weigh significantly less at birth but not thereafter, n = 4-11 per genotype, each time point compared as above.
Supplementary Figure 2 Summary of experimental crosses
Experimental crosses used to determine which compartment of the conceptus (embryo, placental fetal endothelium or placental trophoblast) is the source of maternal circulating DLK1 in pregnancy.
Supplementary Figure 3 Levels of pituitary Gh mRNA do not differ between the groups.
Expression levels of Gh (a) measured by real-time quantitative PCR in pituitaries from females in the cohort. Gh levels did not differ between the groups. Data was normalised to β-actin levels, then expressed as WT = 1. Groups were compared by One-way ANOVA, with Bonferroni's Multiple comparison post-hoc test with WT vs Null and WT x WT vs Null x WT and Null x Null, ***p<0.001.
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Supplementary Figures 1–3 and Supplementary Tables 1–11. (PDF 1167 kb)
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Cleaton, M., Dent, C., Howard, M. et al. Fetus-derived DLK1 is required for maternal metabolic adaptations to pregnancy and is associated with fetal growth restriction. Nat Genet 48, 1473–1480 (2016). https://doi.org/10.1038/ng.3699
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DOI: https://doi.org/10.1038/ng.3699
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