A novel rare missense variant in PLCD1 is associated with recurrent spontaneous abortion and inhibits cell migration and invasion

Recurrent spontaneous abortion (RSA) is a common clinical complication during pregnancy and seriously affects the physical and mental health of women reproductive age. Its etiology and pathogenesis have not yet been fully clarified. In this study, we examined whether genetic variations in PLCD1 contribute to RSA risk. Sanger sequencing was performed to verify PLCD1 (p.H755Y) heterozygous mutation. In silico analysis was used to predict the pathogenicity of this mutation. Wild-type (WT) and mutant-type (MT) plasmids of PLCD1 were constructed and transfected into HTR8/SVneo cells. Western Blot (WB) was used to verify the significant gene overexpression in HTR8/SVneo cells. Transwell migration assays, Matrigel-coated Transwell invasion assays, and cell adhesion experiments were performed using HTR8/SVneo cells transfected with PLCD1 MT, WT and control plasmids respectively. Meanwhile, the expression level of MMP9 was detected via WB. A novel rare heterozygous missense variant in PLCD1 (p.His755Tyr/ c.2263 C > T) was identified in 2 out of 325 (0.6%) RSA patients. This rare variant was absent from in public databases or in 987 control women of Han Chinese ancestry. Functional assays demonstrated that this rare variant significantly inhibited HTR8/SVneo cell migration, invasion, and adhesion, and suppressed MMP9 expression ( p < 0.05). For the first time, we identified a rare heterozygous variant of PLCD1 significantly enriched in patients with RSA. This rare variant may contribute to the pathogenesis of RSA by impairing trophoblast migration and invasion.