Effect of basigin (CD147) on heparin-binding EGF-like growth factor and matrix metalloproteinase expression in human cytotrophoblast cells

AuthorsInstitutions

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

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

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 expressionEndocrinology 2009; 150: 966-976

Mouse Uterus rBSG

ExpressionProfile

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

rBSG

ELISA: HB-EGF, MMPsprotein quantification using cell medium and lysates

Gene Expression Profiling

http://www.allelebiotech.cn/mgr/eWebEditor/UploadFile/20071018225116615.jpg

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rBSG (_g/ml)

MMP-2 protein

concentration (ng/ml)

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rBSG (_g/ml)

HB-EGF protein concentration (ng/ml)

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rBSG (_g/ml)

MMP-9 protein

concentration (ng/ml)

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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 > 41FC

Con

trol

BS

G 0

.5

gB

SG

1.0

g

BS

G 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).