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):

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

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: Background - Basigin in Pregnancy

Figure 1. Expression profiles of MMP-2, MMP-3, MMP-9, and TIMP-3 mRNA during in vitro decidualization. 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 Endocrinology 2009; 150: Mouse Uterus rBSG Expression Profile

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):

Methods - rBSG Treatment, ELISA, and Gene Expression Profiling rBSG ELISA: HB-EGF, MMPs protein quantification using cell medium and lysates Gene Expression Profiling

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. Results - ELISA HB-EGF MMP-2 MMP-9

< - 4 > 4 1 FC Control BSG 0.5  g BSG 1.0  g BSG 5.0  g Results - Gene Expression Profile Figure 2. Heatmap and hierarchical clustering of 2242 features altered by 2-fold. MMP-19

● HB-EGF, MMP-2 and MMP-9 protein levels were not increased in HTR cells following 24 hour treatment with rBSG (Figure 1) – Non-statistically significant increases (1.1 to 1.7 fold) in MMP-9 ● Global gene expression and higherarchial clustering revealed 2,242 features to be altered more than 2 fold (Figure 2) – 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 Results - Summary

● 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 Conclusions

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).