Development of an antagonistic tgf-β by directed blocking of its tβri binding site to inhibition tgf-β signaling

Sepideh Sepehri,1 Reza hasan sajedi,2,* Seyed shahriar arab,3 Mehrdad behmanesh,4

1. Tarbiat Modares University
2. Tarbiat Modares University
3. Tarbiat Modares University
4. Tarbiat Modares University

Abstract

Introduction

Transforming growth factors beta (tgf-βs) play vital roles in regulating cell growth, differentiation and inflammation. in many cancers (for example breast, prostate, colon and lung cancers), autoimmune diseases, cardiovascular diseases and different types of fibrosis, the level of tgf-β and/or tgf-β receptors are abnormally increased. so far tgf-β biological pathways are exploited as therapeutic targets, and several tgf-β signaling inhibitors have been developed, including antibody against, kinase inhibitors, soluble receptors; but no inhibitors have been approved for use in humans. in our previous study, an antagonistic tgf-β variant (anta-tgfβ) designed for inhibition of its signaling pathway. in this study, the biological activity of this variant was checked on mink lung epithelial (mv1lu) cells.

Methods

Designed anta-tgfβ was expressed in shuffle bacteria and purified by ni-nta affinity chromatography. biological activity of purified anta-tgfβ was analyzed on mv1lu cells.

Results

In the present study, anta-tgfβ was expressed and purified. generally, the mature homodimers signal by binding and bringing together two transmembrane receptors, tβri and tβrii, to form heterotetrameric complexes with their receptors i and ii (two of each type). this variant had been designed to contain intact tβrii binding sites and blocked tβri binding sites witch could only bind to tβrii. so, proliferation assay on mv1lu cells was checked in this study. the results showed inhibitory effect of anta-tgfβ on tgf-β activity.

Conclusion

In conclusion, designed antagonist will bind tβrii; however, doesn’t lead to formation of receptor heterotetramer and triggering signal transduction. in addition, designed variant may use as an anti-cancer and anti-fibrotic agent in the future.

Keywords

Transforming growth factor-beta, antagonist tgf-β, cancer, fibrotic disease