TLR4 Modulates Senescence and Paracrine Action in Placental Mesenchymal Stem Cells via Inhibiting Hedgehog Signaling Pathway in Preeclampsia

Abstract

Preeclampsia (PE) is a heterogeneous disease closely associated with the accelerated senescence of the placentas. Placental mesenchymal stem cells (PMSCs) modulate placental development, which is abnormally senescent in PE together with abnormal paracrine. Both pivotal in the placenta development, Toll-like receptor 4 (TLR4) and Hedgehog (HH) pathway are also tightly involved in regulating cellular senescence. This study was aimed at demonstrating that TLR4/HH pathway modulated senescence of placentas and PMSCs in vitro and in vivo. Preeclamptic and normal PMSCs were isolated. Smoothed agonist (SAG) and cyclopamine were used to activate and inhibit HH pathway, respectively. Lipopolysaccharide (LPS) was used to activate TLR4 in vitro and establish the classic PE-like rat model. qRT-PCR, Western blotting, and immunofluorescence were used to detect the expression of TLR4 and HH components (SHH, SMO, and Gli1). Cellular biological function such as proliferation, apoptosis, and migration was compared. Cell cycle analysis, β-galactosidase staining, and the protein expressions of p16 and p53 were detected to analyze the cellular senescence. The secretion levels of human matrix metalloproteinase 9 (MMP-9) and soluble fms-like tyrosine kinase-1 (sFlt-1) were measured in the conditioned medium. Cell migration, invasion, and tube formation were analyzed in HTR8/SVneo cells or human umbilical vein endothelial cells (HUVECs). Our study demonstrated that activation of TLR4 accelerated senescence of PMSCs via suppressing HH pathway both in vitro and in vivo, accompanied by the detrimental paracrine to impair the uterine spiral artery remodeling and placental angiogenesis. Meanwhile, induction of HH pathway could alleviate PE-like manifestations, improve pregnancy outcomes, and ameliorate multiorgan injuries, suggesting that strengthening the HH pathway may serve as a potential therapy in PE.

PFKFB3-mediated glycometabolism reprogramming modulates endothelial differentiation and angiogenic capacity of placenta-derived mesenchymal stem cells

“…PMSCs expressed common MSCs markers (CD44, CD73, CD90, and CD105) but not CD31, CD34, CD45 (Fig. 1 C) [ 38 ].

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Identification of hub genes and pathways associated with cellular senescence in diabetic foot ulcers via comprehensive transcriptome analysis

“…The findings suggest a strong interaction between P53 and TLR proteins, regulating various cellular processes such as DNA repair and replication, CS, differentiation, cell cycle arrest and tumour dynamics 34 . Studies have shown that overexpression of TLR4 can induce CS in osteocytes, placental mesenchymal cells and dental pulp stem cells 35–38 . TLR2 and TLR5, drove CS of MSCs 39 …”

“…34 Studies have shown that overexpression of TLR4 can induce CS in osteocytes, placental mesenchymal cells and dental pulp stem cells. [35][36][37][38] TLR2 and TLR5, drove CS of MSCs. 39 We found eight overlapping hub genes, including TP53, SRC, SIRT1, CCND1, EZH2, CXCL8, AR and CDK4.…”

Non-invasive prediction of preeclampsia using the maternal plasma cell-free DNA profile and clinical risk factors

“…The cfDNA profile in C EPE model contained eight genes, of which pTSS coverages of FOSL2 , CAMKK2 , CCND1 , ITPR1 , PRKACB , WNT7B , CACNB2 and NRF1 genes were stably differential between pregnant women with early-onset PE and healthy controls. CAMKK2 , CCND1 , ITPR1 , PRKACB and WNT7B genes were enriched in more than two pathways associated with PE, including “Hedgehog signaling pathway” ( 35 ), “Hippo signaling pathway” ( 36 ), “AMPK signaling pathway” ( 37 ), “Apelin signaling pathway” ( 38 ), “Autophagy” ( 39 ), “Oxytocin signaling pathway” ( 40 ), “Wnt signaling pathway” ( 41 ) and “VEGFA-VEGFR2 signaling” ( 42 ). The AMPK signaling pathway has been repeatedly reported to be associated with PE based on omics data ( 37 , 43 ).…”