Optogenetic manipulation of BMP signaling to drive chondrogenic differentiation of hPSCs
Optogenetic manipulation of BMP signaling to drive chondrogenic differentiation of hPSCs
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DOI：
10.1016/j.celrep.2023.113502
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发表时间：
2023-11-28
期刊：
影响因子：
8.8
通讯作者：
Kimber，Susan J.
中科院分区：
文献类型：
--
作者：
Humphreys，Paul E. A.;Woods，Steven;Kimber，Susan J.
光遗传学是一项结合光化学、光学和合成生物学来控制细胞行为的快速发展的技术。总之，敏感的光响应光遗传学工具和人类多能干细胞分化模型有可能微调分化，并解开由形态因子控制的细胞规格和组织模式的过程。我们利用光生骨形态发生蛋白（BMP）信号系统（optoBMP）驱动人胚胎干细胞（hESCs）的成软骨分化。我们通过crispr - cas9介导的将optoBMP系统整合到AAVS1位点来设计光敏hESCs。蓝光激活optoBMP，代替BMP生长因子，导致BMP信号机制的激活和软骨表型的上调，与黑暗中的细胞相比，具有显著的转录差异。此外，在光照下分化的细胞可以形成由透明样软骨基质组成的软骨小球。我们的发现表明光遗传学在理解人类发育和组织工程方面的适用性。
Optogenetics is a rapidly advancing technology combining photochemical, optical, and synthetic biology to control cellular behavior. Together, sensitive light-responsive optogenetic tools and human pluripotent stem cell differentiation models have the potential to fine-tune differentiation and unpick the processes by which cell specification and tissue patterning are controlled by morphogens. We used an optogenetic bone morphogenetic protein (BMP) signaling system (optoBMP) to drive chondrogenic differentiation of human embryonic stem cells (hESCs). We engineered light-sensitive hESCs through CRISPR-Cas9-mediated integration of the optoBMP system into the AAVS1 locus. The activation of optoBMP with blue light, in lieu of BMP growth factors, resulted in the activation of BMP signaling mechanisms and upregulation of a chondrogenic phenotype, with significant transcriptional differences compared to cells in the dark. Furthermore, cells differentiated with light could form chondrogenic pellets consisting of a hyaline-like cartilaginous matrix. Our findings indicate the applicability of optogenetics for understanding human development and tissue engineering.