Dinter Teresa, Bocobo Geoffrey A, Yu Paul B
Division of Cardiovascular Medicine, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, MA, USA.
Methods Mol Biol. 2019;1891:221-233. doi: 10.1007/978-1-4939-8904-1_16.
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.
骨形态发生蛋白（BMP）信号通路是转化生长因子β（TGF-β）信号家族的一个子集，由结构多样的受体和配体组成，其组合特异性编码自分泌、旁分泌和内分泌信号，这些信号对于胚胎发育和出生后组织重塑过程中调节组织生长、分化和存活至关重要。这些受体和配体的异常信号传导与多种先天性和后天性疾病有关。可以使用BMP受体激酶的小分子抑制剂来探索各种受体及其配体的作用。已经描述了几种BMP I型受体激酶抑制剂工具化合物，它们表现出足够的选择性，能够在体外或体内区分BMP受体信号传导，在选择性、效力、细胞通透性和药代动力学方面存在各种权衡。本文介绍了几种通过药理学抑制来测定BMP功能的方法。两种体外方法，即BMP介导的SMAD1/5/8磷酸化的细胞内western分析和碱性磷酸酶成骨分化分析，代表了用于测定药理学抑制剂的高效高通量方法。本文还描述了两种体内方法，用于测定BMP信号抑制在胚胎斑马鱼和小鼠发育中的作用。BMP受体激酶的小分子抑制剂是一种重要的补充策略，可用于在生物学和疾病中针对该信号系统的基因功能获得和缺失以及配体陷阱方法。
