Expression of Biglycan in First Trimester Chorionic Villous Sampling Placental Samples and Altered Function in Telomerase-Immortalized Microvascular Endothelial Cells

Amy Chui; Tilini Gunatillake; Shaun P Brennecke; Vera Ignjatovic; Paul T Monagle; John M Whitelock; Dagmar E van Zanten; Jasper Eijsink; Yao Wang; James Deane; Anthony J Borg; Janet Stevenson; Jan Jaap Erwich; Joanne M Said; Padma Murthi

doi:10.1161/ATVBAHA.117.309422

Expression of Biglycan in First Trimester Chorionic Villous Sampling Placental Samples and Altered Function in Telomerase-Immortalized Microvascular Endothelial Cells

Arterioscler Thromb Vasc Biol. 2017 Jun;37(6):1168-1179. doi: 10.1161/ATVBAHA.117.309422. Epub 2017 Apr 13.

Authors

Affiliations

¹ From the Department of Obstetrics and Gynaecology, The University of Melbourne, Parkville, Victoria, Australia (A.C., T.G., S.P.B., P.M.); Sunshine Hospital, St Albans, Victoria, Australia (A.C., T.G., S.P.B., P.M.); Department of Maternal-Fetal Medicine Pregnancy Research Centre, The Royal Women's Hospital, Parkville, Victoria, Australia (S.P.B., A.J.B., J.S., P.M.); Murdoch Children's Research Institute and Department of Clinical Haematology, Department of Paediatrics, The Royal Children's Hospital, The University of Melbourne, Parkville, Victoria, Australia (V.I., P.T.M.); Graduate School of Biomedical Engineering, University of New South Wales, Kensington, Australia (J.M.W.); Maternal Fetal Medicine, Sunshine Hospital, Western Health, St Albans, Victoria, Australia (J.M.S.); Department of Obstetrics and Gynecology, University Medical Centre Groningen, University of Groningen, The Netherlands (D.E.v.Z., J.J.E.); Department of Medicine, School of Clinical Sciences, Monash University, Clayton, Victoria, Australia (Y.W., J.D., P.M.); and The Ritchie Centre, Hudson Institute of Medical Research, Clayton, Victoria, Australia (P.M.). akl.chui83@gmail.com.
² From the Department of Obstetrics and Gynaecology, The University of Melbourne, Parkville, Victoria, Australia (A.C., T.G., S.P.B., P.M.); Sunshine Hospital, St Albans, Victoria, Australia (A.C., T.G., S.P.B., P.M.); Department of Maternal-Fetal Medicine Pregnancy Research Centre, The Royal Women's Hospital, Parkville, Victoria, Australia (S.P.B., A.J.B., J.S., P.M.); Murdoch Children's Research Institute and Department of Clinical Haematology, Department of Paediatrics, The Royal Children's Hospital, The University of Melbourne, Parkville, Victoria, Australia (V.I., P.T.M.); Graduate School of Biomedical Engineering, University of New South Wales, Kensington, Australia (J.M.W.); Maternal Fetal Medicine, Sunshine Hospital, Western Health, St Albans, Victoria, Australia (J.M.S.); Department of Obstetrics and Gynecology, University Medical Centre Groningen, University of Groningen, The Netherlands (D.E.v.Z., J.J.E.); Department of Medicine, School of Clinical Sciences, Monash University, Clayton, Victoria, Australia (Y.W., J.D., P.M.); and The Ritchie Centre, Hudson Institute of Medical Research, Clayton, Victoria, Australia (P.M.).

PMID: 28408374
DOI: 10.1161/ATVBAHA.117.309422

Abstract

Objective: Biglycan (BGN) has reduced expression in placentae from pregnancies complicated by fetal growth restriction (FGR). We used first trimester placental samples from pregnancies with later small for gestational age (SGA) infants as a surrogate for FGR. The functional consequences of reduced BGN and the downstream targets of BGN were determined. Furthermore, the expression of targets was validated in primary placental endothelial cells isolated from FGR or control pregnancies. APPROACH AND RESULTS: BGN expression was determined using real-time polymerase chain reaction in placental tissues collected during chorionic villous sampling performed at 10 to 12 weeks' gestation from pregnancies that had known clinical outcomes, including SGA. Short-interference RNA reduced BGN expression in telomerase-immortalized microvascular endothelial cells, and the effect on proliferation, angiogenesis, and thrombin generation was determined. An angiogenesis array identified downstream targets of BGN, and their expression in control and FGR primary placental endothelial cells was validated using real-time polymerase chain reaction. Reduced BGN expression was observed in SGA placental tissues. BGN reduction decreased network formation of telomerase-immortalized microvascular endothelial cells but did not affect thrombin generation or cellular proliferation. The array identified target genes, which were further validated: angiopoetin 4 (ANGPT4), platelet-derived growth factor receptor α (PDGFRA), tumor necrosis factor superfamily member 15 (TNFSF15), angiogenin (ANG), serpin family C member 1 (SERPIN1), angiopoietin 2 (ANGPT2), and CXC motif chemokine 12 (CXCL12) in telomerase-immortalized microvascular endothelial cells and primary placental endothelial cells obtained from control and FGR pregnancies.

Conclusions: This study reports a temporal relationship between altered placental BGN expression and subsequent development of SGA. Reduction of BGN in vascular endothelial cells leads to disrupted network formation and alterations in the expression of genes involved in angiogenesis. Therefore, differential expression of these may contribute to aberrant angiogenesis in SGA pregnancies.

Keywords: biglycan; cell proliferation; gene expression; pregnancy; thrombosis.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Biglycan / genetics
Biglycan / metabolism*
Case-Control Studies
Cell Line
Chorionic Villi / blood supply*
Chorionic Villi / metabolism*
Chorionic Villi Sampling
Endothelial Cells / metabolism*
Female
Fetal Growth Retardation / genetics
Fetal Growth Retardation / metabolism*
Fetal Growth Retardation / physiopathology
Gene Expression Profiling
Gene Expression Regulation, Developmental
Humans
Infant, Newborn
Infant, Small for Gestational Age
Male
Mice, Inbred C57BL
Microvessels / metabolism*
Neovascularization, Physiologic* / genetics
Pregnancy
Pregnancy Trimester, First / genetics
Pregnancy Trimester, First / metabolism*
RNA Interference
Signal Transduction
Telomerase / genetics
Telomerase / metabolism*
Thrombin / metabolism
Time Factors
Tissue Culture Techniques
Transfection

Substances

BGN protein, human
Biglycan
Telomerase
Thrombin