A locus control region generates distinct active placental lactogen and inactive growth hormone gene domains in term placenta that are disrupted with obesity

Abstract

Introduction

Placental villi include an outer syncytiotrophoblast (STB) layer and an inner layer of cytotrophoblasts (CTBs) that fuse to generate the STB layer in pregnancy. While activation of the locus containing the human (h) placental lactogen (hPL) genes (hPL-A/CSH1 and hPL-B/CSH2) begins in the CTBs, their expression in the STB requires epigenetic modifications and interactions between locus control region (LCR) and gene regulatory sequences. No factor that limits or facilitates hPL LCR/gene interactions for locus activation is reported. The paternally-expressed gene 3 (PEG3/PW1) transcription factor was pursued as a candidate. PEG3 is expressed by villous CTBs but not the STB and putative binding sites were identified in hPL-related sequences.

Methods

PEG3 expression was assessed in multiple cell types including in CTB-like JEG-3 cells. PEG3 binding was also assessed in JEG-3 cells and term placenta samples from women with or without maternal obesity, where chromosomal architecture of the hPL gene locus was also examined.

Results

In JEG-3 cells, PEG3 was found to bind to hypersensitive site (HS III-V) sequences within the LCR. Knockdown of PEG3 in these cells resulted in increased hPL gene expression. In term placenta, PEG3 binding at placenta-specific HS IV was increased with maternal obesity, where a decrease in hPL RNA levels is seen, while PEG3 binding was reduced in women with obesity who develop insulin-treated gestational diabetes mellitus (O/GDM + Ins), where increased hPL gene expression is observed. Chromatin conformation capture revealed distinct hPL gene domain interactions that are modified with maternal obesity but largely reversed in O/GDM + Ins, correlating with PEG3 binding.

Discussion

Decreased PEG3 binding may be required for hPL domain generation and expression during CTB to STB transition.