Department of Pathology and Immunology, Baylor College of Medicine, Houston, Texas, United States of America.
PLoS One. 2012;7(1):e30814. doi: 10.1371/journal.pone.0030814. Epub 2012 Jan 27.
Wnt signaling is crucial for a variety of biological processes, including body axis formation, planar polarity, stem cell maintenance and cellular differentiation. Therefore, targeted manipulation of Wnt signaling in vivo would be extremely useful. By applying chemical inducer of dimerization (CID) technology, we were able to modify the Wnt co-receptor, low-density lipoprotein (LDL)-receptor-related protein 5 (LRP5), to generate the synthetic ligand inducible Wnt switch, iLRP5. We show that iLRP5 oligomerization results in its localization to disheveled-containing punctate structures and sequestration of scaffold protein Axin, leading to robust β-catenin-mediated signaling. Moreover, we identify a novel LRP5 cytoplasmic domain critical for its intracellular localization and casein kinase 1-dependent β-catenin signaling. Finally, by utilizing iLRP5 as a Wnt signaling switch, we generated the Ubiquitous Activator of β-catenin (Ubi-Cat) transgenic mouse line. The Ubi-Cat line allows for nearly ubiquitous expression of iLRP5 under control of the H-2K(b) promoter. Activation of iLRP5 in isolated prostate basal epithelial stem cells resulted in expansion of p63(+) cells and development of hyperplasia in reconstituted murine prostate grafts. Independently, iLRP5 induction in adult prostate stroma enhanced prostate tissue regeneration. Moreover, induction of iLRP5 in male Ubi-Cat mice resulted in prostate tumor progression over several months from prostate hyperplasia to adenocarcinoma. We also investigated iLRP5 activation in Ubi-Cat-derived mammary cells, observing that prolonged activation results in mammary tumor formation. Thus, in two distinct experimental mouse models, activation of iLRP5 results in disruption of tissue homeostasis, demonstrating the utility of iLRP5 as a novel research tool for determining the outcome of Wnt activation in a precise spatially and temporally determined fashion.
Wnt 信号对于多种生物学过程至关重要,包括身体轴的形成、平面极性、干细胞维持和细胞分化。因此,在体内靶向操纵 Wnt 信号将非常有用。通过应用化学二聚体诱导剂(CID)技术,我们能够修饰 Wnt 共受体低密度脂蛋白(LDL)受体相关蛋白 5(LRP5),以产生合成配体诱导的 Wnt 开关 iLRP5。我们表明,iLRP5 寡聚化导致其定位于蓬乱样包含的点状结构和支架蛋白 Axin 的隔离,导致强烈的β-连环蛋白介导的信号转导。此外,我们确定了一个新的 LRP5 细胞质结构域对于其细胞内定位和酪蛋白激酶 1 依赖性β-连环蛋白信号转导至关重要。最后,通过利用 iLRP5 作为 Wnt 信号开关,我们生成了 Ubiquitous Activator of β-catenin(Ubi-Cat)转基因小鼠系。Ubi-Cat 系允许在 H-2K(b)启动子的控制下几乎普遍表达 iLRP5。在分离的前列腺基底上皮干细胞中激活 iLRP5 导致 p63(+)细胞的扩增和重建的小鼠前列腺移植物中的增生。独立地,iLRP5 在成年前列腺基质中的诱导增强了前列腺组织的再生。此外,在雄性 Ubi-Cat 小鼠中诱导 iLRP5 导致前列腺肿瘤进展,从前列腺增生到腺癌,经过几个月。我们还研究了 iLRP5 在 Ubi-Cat 衍生的乳腺细胞中的激活,观察到延长激活导致乳腺肿瘤形成。因此,在两种不同的实验小鼠模型中,激活 iLRP5 导致组织平衡的破坏,证明了 iLRP5 作为一种新的研究工具的实用性,用于以精确的时空方式确定 Wnt 激活的结果。