Placental deficiency of the (pro)renin receptor ((P)RR) reduces placental development and functional capacity

Lachlan G Schofield; Richard G S Kahl; Samantha L Rodrigues; Joshua J Fisher; Saije K Endacott; Sarah J Delforce; Eugenie R Lumbers; Jacinta H Martin; Kirsty G Pringle

doi:10.3389/fcell.2023.1212898

Placental deficiency of the (pro)renin receptor ((P)RR) reduces placental development and functional capacity

Front Cell Dev Biol. 2023 Aug 1:11:1212898. doi: 10.3389/fcell.2023.1212898. eCollection 2023.

Authors

Lachlan G Schofield^{1

2}, Richard G S Kahl^{2

3}, Samantha L Rodrigues^{1

2}, Joshua J Fisher^{2

3}, Saije K Endacott^{1

2}, Sarah J Delforce^{1

2}, Eugenie R Lumbers^{1

2}, Jacinta H Martin^{4

5}, Kirsty G Pringle^{1

2}

Affiliations

¹ School of Biomedical Sciences and Pharmacy, College of Health, Medicine and Wellbeing, University of Newcastle, Callaghan, NSW, Australia.
² Mothers and Babies Research Program, Hunter Medical Research Institute, New Lambton Heights, NSW, Australia.
³ School of Medicine and Public Health, College of Health, Medicine and Wellbeing, University of Newcastle, Callaghan, NSW, Australia.
⁴ School of Environmental and Life Sciences, College of Engineering, Science and Environment, University of Newcastle, Callaghan, NSW, Australia.
⁵ Infertility and Reproduction Research Program, Hunter Medical Research Institute, New Lambton Heights, NSW, Australia.

Abstract

The (pro)renin receptor ((P)RR; also known as ATP6AP2) is a multifunctional receptor. The (P)RR activates the tissue renin-angiotensin system (RAS) and is also involved in regulating integral intracellular pathways such as V-ATPase and Wnt/β-catenin signalling. Given this, the (P)RR may be associated with essential pathways in placentation, however its role within the context of pregnancy remains poorly characterised. The first trimester/extravillous trophoblast cell line, HTR-8/SVneo, underwent an siRNA knockdown where they were incubated for 24 h with a negative control siRNA or siRNA targeting ATP6AP2 mRNA. xCELLigence real-time cell analysis was performed to assess the effect of ATP6AP2 mRNA knockdown on HTR-8/SVneo cell proliferation, migration, and invasion. In subsequent experiments, GFP-encoding lentiviral packaged gene-constructs were used to knockdown (P)RR expression in the trophectoderm of C57/BL6/CBA-F1 mouse blastocysts. Blastocysts were incubated for 6 h with vehicle (no-virus), control virus (non-targeting shRNA and GFP), or (P)RR-knockdown virus ((P)RR shRNA and GFP) before transfer into recipient pseudo-pregnant Swiss CD1 female mice. Fetal and placental tissues were collected and assessed at embryonic age (EA) 10 and 18. (P)RR levels were measured in the labyrinth zone of day 18 placentae and stereological Merz grid analysis was performed to determine the volumetric distribution of trophoblasts, fetal capillaries, and the maternal blood space. We showed that a reduction of ATP6AP2 expression in HTR-8/SVneo cells in vitro, impaired trophoblast proliferation, migration, and invasion. In vivo, decreasing placental labyrinth (P)RR expression adversely effected placental physiology, decreasing placental trophoblast number and total surface area available for exchange, while also increasing maternal blood space. Additionally, decreased (P)RR affected placental efficacy evident by the reduced fetal-placental weight ratio. Our study shows that the (P)RR is necessary for appropriate placental development and function.

Keywords: blastocyst; placenta; placental development; prorenin receptor; trophoblast (TC).

Grants and funding

This work was supported by an NHMRC project grant (APP1161957) and an ARC Future Fellowship awarded to KP (FT150100179).