Chemical genetic screen reveals a role for desmosomal adhesion in mammary branching morphogenesis.
Chemical genetic screen reveals a role for desmosomal adhesion in mammary branching morphogenesis.
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DOI：
10.1074/jbc.m112.411033
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发表时间：
2013-01-25
期刊：
影响因子：
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通讯作者：
Welm BE
中科院分区：
文献类型：
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作者：
Basham KJ;Kieffer C;Shelton DN;Leonard CJ;Bhonde VR;Vankayalapati H;Milash B;Bearss DJ;Looper RE;Welm BE
背景：乳腺分支形态发生是一种高度调控的发育过程，在乳腺癌中常被破坏。结果：原代三维培养中的化学遗传学筛选显示，芳烃受体的激活促进桥粒阻断分支。结论：正常的乳腺分支形态发生需要下调桥粒的表达。意义：桥粒是一种新的机制，通过它暴露于环境污染物可能会影响乳腺发育。在分支形态发生的过程中，乳腺经历了不同的重塑阶段，形成了一个复杂的导管网络，最终产生乳汁并将其输送给新生动物。这些发育事件依赖于对关键细胞通路的严格调控，其中许多通路可能在乳腺癌的发生和发展过程中被破坏。转基因小鼠和体外器官模型以前发现生长因子信号是乳房分支的关键调节因子，但这些通路的下游功能靶点仍不清楚。在这里，我们使用纯化的原代乳腺上皮细胞在成纤维细胞生长因子-2(FGF2)刺激下，在体外模拟乳房分支的形态发生。我们使用了一种正向化学遗传学方法来识别这一过程的调节器，并描述了一种有效的化合物1023，它可以阻断FGF2诱导的分支。在原代乳腺上皮细胞中，我们使用慢病毒介导的芳香烃受体(AHR)的敲除来证明1023通过AHR作用于阻断分支。以1023为工具，我们确定桥粒黏附是AHR信号的一个新靶点，并表明桥粒是AHR激动剂阻断分支的关键。我们的发现支持桥粒在乳腺形态发生和阻止成纤维细胞生长因子诱导的侵袭中的功能作用。
Background: Mammary gland branching morphogenesis is a highly regulated developmental process often disrupted in breast cancer. Results: A chemical genetic screen in primary three-dimensional culture revealed that activation of the aryl hydrocarbon receptor promotes desmosomes to block branching. Conclusion: Down-regulation of desmosomes is required for proper mammary branching morphogenesis. Significance: Desmosomes are a novel mechanism through which exposure to environmental pollutants may affect mammary development. During the process of branching morphogenesis, the mammary gland undergoes distinct phases of remodeling to form an elaborate ductal network that ultimately produces and delivers milk to newborn animals. These developmental events rely on tight regulation of critical cellular pathways, many of which are probably disrupted during initiation and progression of breast cancer. Transgenic mouse and in vitro organoid models previously identified growth factor signaling as a key regulator of mammary branching, but the functional downstream targets of these pathways remain unclear. Here, we used purified primary mammary epithelial cells stimulated with fibroblast growth factor-2 (FGF2) to model mammary branching morphogenesis in vitro. We employed a forward chemical genetic approach to identify modulators of this process and describe a potent compound, 1023, that blocks FGF2-induced branching. In primary mammary epithelial cells, we used lentivirus-mediated knockdown of the aryl hydrocarbon receptor (AHR) to demonstrate that 1023 acts through AHR to block branching. Using 1023 as a tool, we identified desmosomal adhesion as a novel target of AHR signaling and show that desmosomes are critical for AHR agonists to block branching. Our findings support a functional role for desmosomes during mammary morphogenesis and also in blocking FGF-induced invasion.