Identification of selective inhibitors of PTH-receptor for Jansen’s metaphyseal chondrodysplasia

总结 该项目的目的是测试和优化我们最近发现的小非肽分子（重新命名）。 称为皮特分子），以恢复正常的甲状旁腺激素1型受体（PTHR）信号传导和细胞凋亡。 大颞下颌关节（TMJ）髁突形态发生过程中，细胞表达缺陷， 组成型活性受体突变体， 詹森干骺端软骨发育不良（JMC）。基于现有 PI之间的合作（Vilardaga博士，药理学和化学生物学，Taboas博士，部门。口服 和颅面科学，和生物工程）和co-I (Wipf，部门。化学）在U.皮特，一个interdisci- 已经开发了一种多元方法来证明Pitt分子在恢复正常PTHR中的功效 信号转导和细胞形态发生过程，从而更好地了解PTHR信号转导的作用。 髁状突形态发生。 我们提出了2个目标：目标1将确定最有效的变构小分子，用于阻断组成型 使用表达PTHR μ-受体的活细胞中受体信号传导的最新光学分析的PTHR活化 JMC患者中遇到的突变体（PTHR-H223 R、PTHR-T410 P或PTHR-I458 R）和药物化学 优化Pitt分子的阻断特性的方法。目标2将进一步确定有效期， 小分子物质对颞下颌关节髁突形态发生影响的生化和组织学研究 对由来自野生型和小鼠的髁突间充质祖细胞组成的工程化微组织的测定 表达JMC突变体PTHR-H223R。 这项研究计划的意义在于它的前提，以更好地了解PTHR信号的影响， 髁状突形态发生，并为未来的转化研究计划奠定基础， 这些Pitt分子用于治疗颞下颌关节紊乱病的治疗效用。

Summary The objective of this project is to test and optimize our recently identified small non-peptidic molecules (re- ferred to as Pitt molecules) to restore normal parathyroid hormone type 1 receptor (PTHR) signaling and cellu- lar temporomandibular joint (TMJ) condylar morphogenic processes, which are defective in cells expressing constitutive active receptor mutants causing Jansen’s metaphyseal chondrodysplasia (JMC). Based on existing collaborations between PIs (Dr. Vilardaga, Dept. of Pharmacology & Chem. Biol., and Dr. Taboas, Dept. Oral and Craniofacial Sciences, and of Bioengineering) and co-I (Wipf, Dept. of Chemistry) at U.Pitt, an interdisci- plinary approach has been developed to demonstrate the efficacy of Pitt molecules in restoring normal PTHR signaling and cellular morphogenic processes in vitro, and thereby better understand the effects of PTHR sig- naling on condylar morphogenesis. We propose 2 aims: Aim 1 will identify the most effective allosteric small molecules for blocking constitutive PTHR activation using state-of-the-art optical analysis of receptor signaling in live cells expressing PTHR mu- tants encountered in JMC patients (PTHR-H223R, PTHR-T410P, or PTHR-I458R), and medicinal chemistry methods to optimize blocking properties of Pitt molecules. Aim 2 will take a step further by determining the ef- fect of small molecules on temporomandibular joint condylar morphogenesis using biochemical and histological assays on engineered microtissues composed of condylar mesenchymal progenitors from wild-type and mice expressing the JMC mutant PTHR-H223R. The significance of this research program lies in its premise to better understand the effects of PTHR signaling on condylar morphogenesis and lay the groundwork for a future translational research program that examines the therapeutic utility of these Pitt molecules for treatment of temporomandibular disorders.