SP 5: Characterization of Fetal Endothelial Cells in Relation to Maternal Environment

Research questions, aims

Preexisting diseases of the mother may affect in utero development of the fetus and long-term programming. This subprogram will investigate gene and protein expression as well as functional parameters of placental and fetal endothelial cells derived from different maternal environments. We aim to compare fetal endothelial cells between pregnancies of healthy women and women with risk factors (diabetes, obesity, smoking, age), PE or in vitro fertilization. We expect functionally relevant differences in gene or protein expression, which will help to clarify molecular mechanisms of pregnancy complications and long-term consequences of fetal exposure to maternal factors. Moreover, we aim to identify targets of predictive value for chronic diseases of the offspring.

Scientific background

PE programs fetal vasculature in utero but underlying mechanisms are incompletely understood. A recent study compared human umbilical vein endothelial cells (HUVEC) from normotensive and preeclamptic pregnancies and identified >1,000 differentially expressed genes with specific patterns for male and female cells (1). Many of the PE-dysregulated genes are associated with endothelial dysfunction suggesting that they contribute to increased cardiovascular risk in the offspring. However, the functional relevance of gene dysregulation and the effect of individual maternal risk factors on fetal endothelial cells have not been studied.

Own preceding work

The Heller lab has a long-standing experience in endothelial cell biology with specific interest in stress signaling and adaptation and metabolic processes involved therein. We described the role of the metabolic enzyme AMP-activated protein kinase (AMPK), in endothelial cells including its crosstalk with several signaling pathways, its role in angiogenesis and its alterations in cellular stress situations, senescence and aging (2-4). Mechanistically, we focus on the role of posttranslational protein modifications as key mechanisms of cellular dysfunction and ageing. In this context, we have recently described the endothelial cell glycome and related it to cellular functions (5). We have comprehensively characterized endothelial cell metabolism in senescence and inflammation (6).

Method spectrum, involvement of the Medical Scientist

The Heller lab routinely isolates HUVEC and endothelial cells from other tissues. All methods to characterize endothelial function are established (mRNA, protein and metabolite analyses, functional assays (migration, angiogenesis), metabolic assays (flux measurements, Seahorse analyses), analysis of protein modifications, omics technologies). The Medical Scientist to be employed will profit from this spectrum and work in close collaboration with subprogram 1 and 4 to get access to tissue samples from specific cohorts for endothelial cell isolation. She/he will also have access to the endometrium biobank (see subprogram 1) to analyze samples for specific markers of endothelial dysfunction identified in cell studies.

Relevance for CEPRE and for Reproductive Health

The Medical Scientist will further clarify mechanisms through which maternal factors affect fetal health. The expected data may help to identify biomarkers for determining the risk of offspring to develop chronic disease.

References

1. Zhou C, Yan Q, Zou QY, Zhong XQ, Tyler CT, Magness RR, Bird IM, Zheng J. Sexual Dimorphisms of Preeclampsia-Dysregulated Transcriptomic Profiles and Cell Function in Fetal Endothelial Cells. Hypertension. 2019;74(1):154-63.
2. Stahmann N, Woods A, Spengler K, Heslegrave A, Bauer R, Krause S, Viollet B, Carling D, Heller R. Activation of AMP-activated protein kinase by vascular endothelial growth factor mediates endothelial angiogenesis independently of nitric-oxide synthase. J Biol Chem. 2010;285(14):10638-52.
3. Zibrova D, Vandermoere F, Goransson O, Peggie M, Marino KV, Knierim A, Spengler K, Weigert C, Viollet B, Morrice NA, Sakamoto K, Heller R. GFAT1 phosphorylation by AMPK promotes VEGF-induced angiogenesis. Biochem J. 2017;474(6):983-1001.
4. Spengler K, Kryeziu N, Grosse S, Mosig AS, Heller R. VEGF Triggers Transient Induction of Autophagy in Endothelial Cells via AMPKalpha1. Cells. 2020;9(3).
5. Di Sanzo S, Spengler K, Leheis A, Kirkpatrick JM, Randler TL, Baldensperger T, Dau T, Henning C, Parca L, Marx C, Wang ZQ, Glomb MA, Ori A, Heller R. Mapping protein carboxymethylation sites provides insights into their role in proteostasis and cell proliferation. Nat Commun. 2021;12(1):6743.
6. Stabenow LK, Zibrova D, Ender C, Helbing DL, Spengler K, Marx C, Wang ZQ, Heller R. Oxidative Glucose Metabolism Promotes Senescence in Vascular Endothelial Cells. Cells. 2022;11(14).