Mechanisms of WNT signaling in bone

描述（由申请人提供）：Wnt信号传导为开发新型骨合成代谢药物提供了一种有前景的靶向途径。迄今为止，大多数研究支持成骨细胞系细胞中自分泌或旁分泌Wnt信号直接控制成骨细胞生物学的模型。然而，这一模型受到了最近一项研究的挑战，该研究得出结论，LRP 5是Wnt蛋白的辅助受体，不直接在成骨细胞中发挥作用，而是通过调节5-羟色胺的肠道产生。因此，这项研究对骨中直接Wnt信号传导的生理相关性产生了不确定性。不确定性的一个主要原因是，2.3Col1-Cre在成骨细胞中的<$-catenin（经典Wnt信号传导的强制性效应子）的遗传缺失并不影响出生后动物的成骨细胞数量或功能。然而，先前在小鼠胚胎中的研究表明，Wnt/Lrp 5/<$-catenin信号可能在2.3Col1-Cre变得活跃之前的一个阶段起作用。由于缺乏适当的遗传工具，在产后生活中直接测试这一概念是不可行的。我们现在已经开发了一种新的Tet-on系统，该系统允许对出生后小鼠的骨祖细胞进行基因操作。因此，我们建议在出生后删除骨祖细胞中的<$-连环蛋白，以检验<$-连环蛋白直接调节出生后骨形成的假设（目的1）。该领域进展的第二个关键障碍是缺乏对成骨细胞系中介导Wnt功能的分子机制的理解。研究一直受到缺乏一个强大的小鼠模型，其中Wnt蛋白可以被操纵和评估其在体内的急性信号传导能力的阻碍。我们现在已经开发了这样的模型，其中有效的骨合成代谢Wnt配体可以以受控的方式被激活。因此，在目标2和3中，我们将采用这种新的小鼠模型来研究Wnt 7 b在体内诱导骨形成的生物化学和遗传学信号转导机制。 公共卫生相关性：需要新的策略来安全地促进骨形成以治疗骨质疏松症。已知Wnt信号传导刺激骨形成，并为开发新的骨合成代谢剂提供了有前途的靶向途径，但其潜在的分子机制尚不清楚。该提案旨在了解Wnt蛋白的强效骨刺激功能的机制。本研究结果将为开发新型骨增强药物提供分子框架。

DESCRIPTION (provided by applicant): Wnt signaling provides a promising target pathway for developing novel bone anabolic agents. Most studies to date have supported a model in which autocrine or paracrine Wnt signaling in osteoblast-lineage cells directly controls osteoblast biology. This model however, was challenged by a recent study that concluded that LRP5, a co-receptor for Wnt proteins, does not function directly in osteoblasts, but rather through regulating enteric production of serotonin. This study therefore has cast uncertainty about the physiological relevance of direct Wnt signaling in bone. A major cause for the uncertainty is that genetic deletion of ¿-catenin (an obligatory effector of canonical Wnt signaling) in osteoblasts by 2.3Col1-Cre did not affect osteoblast number or function in postnatal animals. However, previous work in the mouse embryo indicates that Wnt/Lrp5/¿-catenin signaling may function at a stage before 2.3Col1-Cre becomes active. Directly testing this notion in postnatal life has not been feasible because of the lack of proper genetic tools. We have now developed a novel Tet-on system that allows for gene manipulation in osteoprogenitors specifically in postnatal mice. Therefore, we propose to delete ¿-catenin in osteoprogenitors postnatally to test the hypothesis that ¿-catenin directly regulates bone formation in postnatal life (Aim 1). A second critical barrier to progress in the field is the lack of understanding of the molecular mechanisms that mediate Wnt function in osteoblast-lineage cells. Research has been hindered by the lack of a robust mouse model in which a Wnt protein can be manipulated and assessed for its acute signaling ability in vivo. We have now developed such a model wherein a potent bone anabolic Wnt ligand can be activated in a controlled manner. Therefore, in Aims 2 and 3, we will employ this new mouse model to investigate both biochemically and genetically the signal transduction mechanisms through which Wnt7b induces bone formation in vivo. PUBLIC HEALTH RELEVANCE: Novel strategies are required to safely promote bone formation to treat osteoporosis. Wnt signaling has been known to stimulate bone formation and provides a promising target pathway for developing novel bone anabolic agents, but the underlying molecular mechanisms are not well understood. This proposal is designed to understand the mechanism responsible for the potent bone-stimulating function of Wnt proteins. Research results from this study will provide a molecular framework for developing novel bone-enhancing pharmaceutics.