Trophoblast H2S Maintains Early Pregnancy via Regulating Maternal-Fetal Interface Immune Hemostasis

Abstract

Context Dysregulated immune hemostasis occurs in unexplained recurrent spontaneous abortion (URSA). Synthesized by cystathionine β-synthase (CBS) and cystathionine γ-lyase (CSE), hydrogen sulfide (H2S) promotes regulatory T-cell differentiation and regulates immune hemostasis; yet, its role in URSA is elusive. Objective To determine if H2S plays a role in early pregnancy and if dysregulated H2S signaling results in RSA. Design First-trimester placenta villi and decidua were collected from normal and URSA pregnancies. Protein expression were examined by immunohistochemistry and immunoblotting. Human trophoblast HTR8/SVneo and JEG3 cells were treated with H2S donors; HTR8/SVneo cells were transfected with CBS RNAi or cDNA. Cell migration and invasion were determined by trans-well assays; trophoblast transcriptomes were determined by RNA-seq. Wild-type, CBS-deficient, and CBA/J × DBA/2 mice were treated with CBS and CSE inhibitors or H2S donors to determine the role of H2S in early pregnancy in vivo. Results CBS and CSE proteins showed cell-specific expressions but only CBS decreased in the villous cytotrophoblast in URSA vs. normal subjects. H2S donors promoted migration and invasion and MMP-2 and VEGF expression in HTR8/SVneo and JEG3 cells, similar to forced CBS expression in HTR8/SVneo cells. The CBS-responsive transcriptomes in HTR8/SVneo cells contained differentially regulated genes, i.e., IL-1R and PGTS2, that are associated with nuclear factor-κB-mediated inflammatory response. In vivo, dysregulated CBS/H2S signaling significantly increased embryonic resorption and decidual Th1/Th2 imbalance in mice, which was partially rescued by H2S donors. Conclusion CBS/H2S signaling maintains early pregnancy possibly via regulating maternal-fetal interface immune hemostasis, offering opportunities for H2S-based immunotherapies for URSA.

Circ-CCNB1 Modulates Trophoblast Proliferation and Invasion in Spontaneous Abortion by Regulating miR-223/SIAH1 axis

“…JEG3 is a choriocarcinoma, and HTR8/SVneo is an extravillous trophoblast cell line. JEG-3 and HTR8/SVneo cells have been widely used as a model for placental trophoblast, as well as modeling the physiologically invasive extravillous trophoblast in spontaneous abortion and early pregnancy ( 36 , 37 ). CircRNAs can regulate gene expression by influencing transcription, mRNA turnover, and translation by sponging RNA-binding proteins and microRNAs ( 38 ).…”

Focal adhesion kinase signaling is necessary for the hydrogen sulfide-enhanced proliferation, migration and invasion of HTR8/SVneo human trophoblasts

“…After 24 hours of culture, HTR8/SVneo cells were lysed in 200 μL of RIPA lysis solution supplemented with protease inhibitor and phosphatase inhibitor cocktail (1:100) on ice for 10 minutes, and the proteins in the supernatant were collected by centrifugation at 10,000× g for 10 minutes at 4°C. Protein samples (2 mg/mL) were separated by 10% SDS-PAGE and transferred to polyvinylidene fluoride membranes (0.45 μm, Millipore) as described previously [13,20] . After blocking, the membrane was probed with rabbit monoclonal antibodies against p-FAK (1:1000; Signalway Antibody, cat.…”

Enhanced Sp1/YY1 Expression Directs CBS Transcription to Mediate VEGF-Stimulated Pregnancy-Dependent H 2 S Production in Human Uterine Artery Endothelial Cells

“…Our recent work has shown that enhanced H 2 S production via selective CBS upregulation contributes to pregnancy-associated uterine vasodilation and endometrial/placental angiogenesis, 9,11,20,21 as well as immune tolerance at the maternal-fetal interface 34 and that pregnancy augments VEGF-stimulated human artery endothelial cell H 2 S production via selective CBS upregulation. 19 Adenoviral delivery of VEGF gene abelites undernutrition-induced fetal growth restriction via upregulating UtBF.…”