Pharmacologic Strategies for Assaying BMP Signaling Function.

The bone morphogenetic protein (BMP) signaling pathway, a subset of the transforming growth factor β (TGF-β) signaling family, consists of structurally diverse receptors and ligands whose combinatorial specificity encodes autocrine, paracrine, and endocrine signals essential for regulating tissue growth, differentiation, and survival during embryonic patterning and postnatal tissue remodeling. Aberrant signaling of these receptors and ligands is implicated in a variety of inborn and acquired diseases. The roles of various receptors and their ligands can be explored using small molecule inhibitors of the BMP receptor kinases. Several BMP type I receptor kinase inhibitor tool compounds have been described that exhibit sufficient selectivity to discriminate BMP receptor signaling in vitro or in vivo, with various trade-offs in selectivity, potency, cell permeability, and pharmacokinetics. Several methods for assaying BMP function via pharmacologic inhibition are presented. Two in vitro methods, an In-Cell Western assay of BMP-mediated SMAD1/5/8 phosphorylation and an alkaline phosphatase osteogenic differentiation assay, represent efficient high-throughput methodologies for assaying pharmacologic inhibitors. Two in vivo methods are described for assaying the effects of BMP signaling inhibition in embryonic zebrafish and mouse development. Small molecule inhibitors of BMP receptor kinases represent an important complementary strategy to genetic gain- and loss-of-function and ligand-trap approaches for targeting this signaling system in biology and disease.