Effect of proline rich 15-deficiency on trophoblast viability and survival

PLoS One. 2017 Apr 5;12(4):e0174976. doi: 10.1371/journal.pone.0174976. eCollection 2017.

Abstract

Deviations from the normal program of gene expression during early pregnancy can lead to early embryonic loss as well as dysfunctional placentation, which can cause significant morbidity and mortality. Proline rich 15 (PRR15) is a low molecular weight nuclear protein expressed by the trophoblast during early gestation. Lentivirus-mediated knockdown of PRR15 mRNA in ovine trophectoderm led to demise of the embryo by gestational day 15, providing compelling evidence that PRR15 expression is critical during this precarious window of development. Our objective was to determine the effect of PRR15 knockdown on trophoblast gene expression, proliferation, and survival. The first-trimester human trophoblast cell line, ACH-3P, was infected with control lentivirus or a lentivirus expressing a short hairpin (sh)RNA to target PRR15 mRNA for degradation, resulting in a 68% reduction in PRR15 mRNA. Microarray analysis of these cell lines revealed differential expression of genes related to cancer, focal adhesion, and p53 signaling. These changes included significant up-regulation of GDF15, a cytokine increased in pregnancies with preeclampsia. Viability and proliferation decreased in PRR15-deficient cells, which was consistent with down-regulation of cell cycle-related genes CCND1 and CDK6 and an up-regulation of CCNG2 and CDKN1A in the PRR15-deficient cells. TNFSF10, a tumor necrosis factor superfamily member known to induce apoptosis increased significantly in the PRR15-deficient cells. Migration through a basement membrane matrix decreased and an increased population of apoptotic cells was present when treated with shRNA to target PRR15. These results suggest that PRR15 enhances trophoblast viability and survival during early implantation and placentation.

MeSH terms

  • Apoptosis / genetics
  • Blotting, Western
  • Cell Cycle / genetics
  • Cell Line
  • Cell Proliferation / genetics
  • Gene Expression Regulation, Developmental / genetics
  • Gene Knockdown Techniques
  • Humans
  • Nuclear Proteins / isolation & purification
  • Oligonucleotide Array Sequence Analysis
  • Proteins / physiology*
  • RNA, Small Interfering / genetics
  • Real-Time Polymerase Chain Reaction
  • Trophoblasts / physiology*

Substances

  • Nuclear Proteins
  • Proteins
  • RNA, Small Interfering
  • proline rich 15 protein, human

Grant support

This work was supported by Agriculture and Food Research Initiative Competitive Grant no. 2009-65203-05670 from the United States Department of Agriculture National Institute of Food and Agriculture. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.