Optogenetic control of the Bone Morphogenetic Protein signalling pathway through engineered blue light-sensitive receptors
Optogenetic control of the Bone Morphogenetic Protein signalling pathway through engineered blue light-sensitive receptors
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DOI：
10.1101/2020.04.27.063073
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发表时间：
2020-04
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影响因子：
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通讯作者：
Paul A Humphreys;Steven Woods;Christopher A Smith;S. Cain;Robert J. Lucas;S. Kimber
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作者：
Paul A Humphreys;Steven Woods;Christopher A Smith;S. Cain;Robert J. Lucas;S. Kimber
骨形态发生蛋白（BMP）是转化生长因子β（TGFβ）超家族的成员，在发育过程中起着至关重要的作用;包括肾、肝和骨骼组织的中胚层图案化和特化。体外发育模型目前依赖于昂贵且不可靠的重组BMP蛋白，其不能动态或精确地干扰BMP信号传导途径。在这里，我们开发了一种新的光遗传学BMP信号系统（optoBMP），能够通过蓝光照射快速诱导经典的BMP信号通路。我们证明了optoBMP系统在多种人类细胞系中的效用，以通过SMAD 1/5的磷酸化和核转位启动信号转导，导致BMP靶基因的上调，包括DNA结合ID 2和ID 4的抑制剂。此外，我们展示了如何optoBMP系统可用于微调激活的BMP信号通路，通过可变的光刺激。BMP信号传导的光遗传学控制将使得能够在细胞和发育系统中的各种应用中进行动态和高通量干预。
Bone Morphogenetic Proteins (BMPs) are members of the Transforming Growth Factor β (TGFβ) superfamily and have crucial roles during development; including mesodermal patterning and specification of renal, hepatic and skeletal tissues. In vitro developmental models currently rely upon costly and unreliable recombinant BMP proteins that do not enable dynamic or precise perturbation of the BMP signalling pathway. Here, we develop a novel optogenetic BMP signalling system (optoBMP) that enables rapid induction of the canonical BMP signalling pathway through illumination with blue light. We demonstrate the utility of the optoBMP system in multiple human cell lines to initiate signal transduction through phosphorylation and nuclear translocation of SMAD1/5, leading to upregulation of BMP target genes including Inhibitors of DNA binding ID2 and ID4. Furthermore, we demonstrate how the optoBMP system can be used to fine-tune activation of the BMP signalling pathway through variable light stimulation. Optogenetic control of BMP signalling will enable dynamic and high-throughput intervention across a variety of applications in cellular and developmental systems.