Title

Identification of Commercially Available Antibodies That Block Ligand-Binding by BMPR2

Authors

Document Type

Poster

Publication Date

11-9-2018

Abstract

The Bone Morphogenetic Protein (BMP) signaling pathway comprises the largest subdivision of the Transforming Growth Factor (TGFβ) superfamily. BMP signaling plays essential roles in both embryonic development and postnatal tissue homeostasis. When dysregulated, BMP signaling underlies numerous human pathologies ranging from pulmonary arterial hypertension to heterotopic ossification. Thus, understanding the basic mechanisms by which BMP signaling occurs and is regulated is a highly important goal and may yield translational opportunities. Unfortunately, there are limited tools available to perform functional evaluation of this pathway and genetic approaches are frequently confounded by developmental requirements or ability of pathway components to compensate for one another. While significant progress has been made in developing pharmacological inhibitors for type 1 BMP receptors, specific inhibitors of type 2 receptor are poorly represented. For these reasons, we sought to identify and validate commercially available antibodies that neutralize the ligand-binding function of the Bone Morphogenetic Protein Receptor Type 2 (BMPR2) extracellular domain (ECD). Using a modified, cell-free immunoprecipitation assay quantified by ELISA, we examined the neutralizing ability of the mouse monoclonal antibody 3F6 and found a dose-dependent inhibition of BMPR2-ECD ligand-binding. We extended the results by examining the ability of 3F6 to block endogenous BMPR2 function in a BMP-responsive cell line and found that, consistent with the results of our cell-free system, pre-treatment of HEK293T cells with 3F6 leads to reduced sensitivity in response to BMP pathway activation by BMP2. These results provide important proof-of-concept data for future studies interrogating BMPR2 function in numerous physiological and pathophysiological contexts.

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