A Microphysiological Approach to Evaluate Effectors of Intercellular Hedgehog Signaling in Development.
A Microphysiological Approach to Evaluate Effectors of Intercellular Hedgehog Signaling in Development.
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DOI：
10.3389/fcell.2021.621442
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发表时间：
2021
影响因子：
5.5
通讯作者：
Lipinski RJ
中科院分区：
文献类型：
--
作者：
Johnson BP;Vitek RA;Morgan MM;Fink DM;Beames TG;Geiger PG;Beebe DJ;Lipinski RJ
组织微环境中的旁分泌信号是形态发生的中心介质，体外模拟这种动态细胞间活动对于理解正常和异常发育至关重要。例如，Sonic Hedgehog （Shh）信号是涉及多个发育过程的保守机制，与人类出生缺陷（包括唇腭裂）密切相关。SHH配体由上皮外胚层产生、加工和分泌，通过细胞外基质穿梭，在细胞外基质中与间充质受体结合，建立转录反应梯度，驱动口面部形态发生。在人类中，遗传易感性和作用于不同分子靶点的环境损害的复杂相互作用被认为是口腔面部裂病因的基础。因此，需要一种易于处理的体外方法来模拟这种复杂的细胞和环境相互作用，并对多步骤信号级联的破坏敏感。我们开发了一种基于微板的设备，它支持上皮直接覆盖在细胞外基质嵌入的间质上，模拟了发育中的口面部组织的基本组织结构。上皮产生的SHH配体在邻近的间质中产生SHH驱动的转录梯度，再现了在体内观察到的途径活性梯度。Shh通路激活可被上皮分泌、细胞外基质转运和间充质感应靶点的小分子抑制剂拮抗，支持该方法在完整Shh通路的高含量化学筛选中使用。总之，这些发现证明了一种新颖实用的微生理模型，在研究上皮-间质相互作用和发育过程中的环境信号干扰方面具有广泛的实用性。
Paracrine signaling in the tissue microenvironment is a central mediator of morphogenesis, and modeling this dynamic intercellular activity in vitro is critical to understanding normal and abnormal development. For example, Sonic Hedgehog (Shh) signaling is a conserved mechanism involved in multiple developmental processes and strongly linked to human birth defects including orofacial clefts of the lip and palate. SHH ligand produced, processed, and secreted from the epithelial ectoderm is shuttled through the extracellular matrix where it binds mesenchymal receptors, establishing a gradient of transcriptional response that drives orofacial morphogenesis. In humans, complex interactions of genetic predispositions and environmental insults acting on diverse molecular targets are thought to underlie orofacial cleft etiology. Consequently, there is a need for tractable in vitro approaches that model this complex cellular and environmental interplay and are sensitive to disruption across the multistep signaling cascade. We developed a microplate-based device that supports an epithelium directly overlaid onto an extracellular matrix-embedded mesenchyme, mimicking the basic tissue architecture of developing orofacial tissues. SHH ligand produced from the epithelium generated a gradient of SHH-driven transcription in the adjacent mesenchyme, recapitulating the gradient of pathway activity observed in vivo. Shh pathway activation was antagonized by small molecule inhibitors of epithelial secretory, extracellular matrix transport, and mesenchymal sensing targets, supporting the use of this approach in high-content chemical screening of the complete Shh pathway. Together, these findings demonstrate a novel and practical microphysiological model with broad utility for investigating epithelial-mesenchymal interactions and environmental signaling disruptions in development.
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影响因子：
3.7
作者：
Belair DG;Wolf CJ;Wood C;Ren H;Grindstaff R;Padgett W;Swank A;MacMillan D;Fisher A;Winnik W;Abbott BD
通讯作者：
Abbott BD
影响因子：
6.5
作者：
Chevy, F;Illien, F;Roux, C
通讯作者：
Roux, C
影响因子：
10.4
作者：
Everson, Joshua L.;Sun, Miranda R.;Lipinski, Robert J.
通讯作者：
Lipinski, Robert J.
影响因子：
4.6
作者：
Hu, Diane;Young, Nathan M.;Marcucio, Ralph S.
通讯作者：
Marcucio, Ralph S.
DOI：
10.1002/bdra.23021
发表时间：
2012-07
影响因子：
--
作者：
Carmichael, Suzan L.;Shaw, Gary M.;Lammer, Edward J.
通讯作者：
Lammer, Edward J.