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Effect of basigin (CD147) on heparin-binding EGF-like growth factor and matrix metalloproteinase expression in human cytotrophoblast cells Authors Institutions Background - HB-EGF, MMPs ● ● Trophoblast cell survival is dependent on matrix metaloproteinase (MMP) mediated shedding HB-EGF Basigin is an Ig superfamily cell surface glycoprotein with multiple functions, including induction of MMPs Mol Reprod Dev 2009, 76(12): 1116-1127 Background - What can we learn from Basigin null mice ? ● Severely impaired implantation – ● ● ● ● Normal embryos, but <8% achieved implantation Females are capable of supporting Basigin wildtype embryos, but with severely reduced efficiency Males experience spermatogenesis arrest at M1 metaphase Impaired neutrophil migration into renal tissue after ischemia/reperfusion (renal injury) Deficient MCT1, MCT3, MCT4 in retina Background - Basigin in Pregnancy Figure 1. Basigin expression in implantation sites of the mouse uterus on d 5 (A), d 6 (B), d 7 (C), and d 8 (D) of pregnancy Endocrinology 2009, 150: 966-976 Mouse Uterus rBSG Expression Profile Figure 1. Expression profiles of MMP-2, MMP-3, MMP-9, and TIMP-3 mRNA during in vitro decidualization. Endocrinology 2009; 150: 966-976 Figure 2. Treatment of uterine stromal cells with rBSG increases MMP-3 and MMP-9 mRNA levels without affecting MMP-2 or TIMP-3 expression Objective: To determine the effect of basigin on HB-EGF and MMP expression in human first trimester cytotrophoblast cells. ? BSG ? Mol Reprod Dev 2009, 76(12): 1116-1127 Methods - rBSG Treatment, ELISA, and Gene Expression Profiling ELISA: HB-EGF, MMPs protein quantification using cell medium and lysates rBSG Gene Expression Profiling http://www.allelebiotech.cn/mgr/eWebEditor/UploadFile/20071018225116615.jpg B 2.5 MMP-2 Results - ELISA 0.08 MMP-2 protein concentration (ng/ml) A 2 HB-EGF 0.07 1.5 1 HB-EGF protein concentration (ng/ml) 0.06 0.5 0.05 0.04 0 0 0.03 0.5 1.0 5.0 rBSG (μg/ml) 0.02 C 2 0.01 MMP-9 1.8 0 1.6 0.5 1.0 5.0 rBSG (μg/ml) Figure 1. rBSG does not stimulate (A) HB-EGF, (B) MMP-2 and (C) MMP-9 protein expression from HTR cells. Cells were treated with rBSG at 0 (control), 0.1, 1, and 5 µg/ml for 24 h as described, and the cell lysates and MMP-2 were collected. Data expressed as mean from separate experiments. MMP-9 protein concentration (ng/ml) 0 1.4 1.2 1 0.8 0.6 0.4 0.2 0 0 0.5 1.0 rBSG (μg/ml) 5.0 Control BSG 0.5 g BSG 1.0 g BSG 5.0 g Results - Gene Expression Profile MMP-19 <-4 1 FC >4 Figure 2. Heatmap and hierarchical clustering of 2242 features altered by 2-fold. Results - Summary ● HB-EGF, MMP-2 and MMP-9 protein levels were not increased in HTR cells following 24 hour treatment with rBSG (Figure 1) – ● Global gene expression and higherarchial clustering revealed 2,242 features to be altered more than 2 fold (Figure 2) – ● Non-statistically significant increases (1.1 to 1.7 fold) in MMP-9 MMP-19 was found to be altered by more than 2 fold Protein analysis revealed 60 genes to have more than 10 protein-protein interactions Conclusions ● ● Effect of Basigin on human trophoblast cells – HB-EGF expression - No Effect – Protein and gene expression - Increased MMP-19 Further research is required to better understand the role of MMP-19 in HTR cells Thank you for your attention, I am now ready for questions! This work was supported by a VARI-GRMERC Medical Student Summer Research Internship (DJH) and National Institutes of Health Grant U54 HD40093 (REL).