Small molecule control of Wnt signal transduction

Wnt信号转导的小分子控制

• 批准号: BB/I021922/1
• 负责人: Karen Liu
• 金额: $ 47.95万
• 依托单位: King's College London
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2012
• 资助国家: 英国
• 起止时间: 2012 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FI021922%2F1
• 关键词: Small; molecule; control; Wnt; signal

Studying the functions of specific proteins by inactivation within an intact animal presents several challenges. Genetic deletion, or 'knockout,' technology completely eliminates a protein, but since the protein may have roles in different tissues or at different stages of development, a knockout mouse may not survive to the desired stage of maturity. Drug-based approaches are attractive alternatives because small molecules can be used to inhibit protein function in a genetically normal animal, they can be administered and removed at specific times and are thus reversible, and they often provide attractive lead compounds for drug development. However, small molecules present their own challenges. Is there a small molecule that targets the protein of interest? Can it be delivered to a live animal? Most importantly, can off-target effects of the small molecule be minimized? To study the function of signaling proteins in development we are combining the advantages of gene targeting and small molecules, using a novel approach called inducible stabilization in which a non-toxic drug regulates the stability of any specific protein of interest. As an embryo develops and grows, each cell must be precisely coordinated with its neighbors in order for the animal to be properly patterned. These cells must be communicating with surrounding tissues and making cell fate decisions at all times. How do cells know which stimuli to respond to and which stimuli to ignore? A more thorough understanding of what key signaling molecules are doing in specific types of cells will give us a better understanding of how an animal is built, as well as what happens when development goes awry. My work aims to address these questions by adapting novel chemical tools to help us better understand embryonic development. A major problem when studying developmental processes is that these processes occur over time. For example, first the embryo makes neural precursors, then it allocates some of these cells to become different types of neural tissue. Meanwhile, because the embryo is growing and changing in shape, all these tissues need to develop and be moved to the right place at the right time. Somehow the cells can sense an 'architectural plan' and coordinate to make brains in the head and motor neurons precisely where the limbs are developing. Specifically, this work will focus on making new tools to study beta-catenin, a molecule that is important in development and in diseases such as cancer. Because of the importance of this molecule, we are using it as a 'test case' for these new technologies. In this way, our 'test case' will teach us a great deal about these new methods and will also likely be generally useful for future biological studies. A second aim makes use of our existing drug-dependent allele of glycogen synthase kinase-3 (GSK-3) to study neural crest migration. This protein is also an important regulator of development and disease. We have previously shown, using similar methods, that GSK-3 is necessary during a critical period in palate development. Using these mice, we found that adding back this protein during mid-gestation was sufficient to rescue cleft palate in mutant mice. We hope to use these kinds of approaches to understand the timing and amounts of gene activity required in different developmental processes. Taken together, this project will provide important new chemical biology tools for the research community as well as gaining insight into molecular mechanisms underlying neural crest migration. A better understanding of neural crest migration will likely help us better understand human development and diseases processes such as cancer metastasis.

通过在完整动物体内失活来研究特定蛋白质的功能面临着一些挑战。基因删除或“基因敲除”技术完全消除了蛋白质，但由于该蛋白质可能在不同组织或不同发育阶段发挥作用，因此基因敲除小鼠可能无法存活到所需的成熟阶段。基于药物的方法是有吸引力的替代方案，因为小分子可用于抑制遗传正常动物的蛋白质功能，它们可以在特定时间施用和去除，因此是可逆的，并且它们通常为药物开发提供有吸引力的先导化合物。然而，小分子也面临着自己的挑战。是否存在针对目标蛋白质的小分子？它可以传递给活体动物吗？最重要的是，小分子的脱靶效应能否最小化？为了研究信号蛋白在发育过程中的功能，我们结合了基因靶向和小分子的优点，使用了一种称为诱导稳定的新方法，其中无毒药物调节任何特定目标蛋白的稳定性。随着胚胎的发育和生长，每个细胞必须与其相邻细胞精确协调，以便动物形成正确的模式。这些细胞必须始终与周围组织沟通并做出细胞命运决定。细胞如何知道对哪些刺激做出反应以及忽略哪些刺激？更全面地了解关键信号分子在特定类型细胞中的作用将使我们更好地了解动物是如何构建的，以及当发育出现问题时会发生什么。我的工作旨在通过采用新颖的化学工具来解决这些问题，以帮助我们更好地了解胚胎发育。研究发育过程时的一个主要问题是这些过程随着时间的推移而发生。例如，胚胎首先产生神经前体，然后将其中一些细胞分配成不同类型的神经组织。同时，由于胚胎正在生长和形状发生变化，所有这些组织都需要发育并在正确的时间移动到正确的位置。不知何故，这些细胞可以感知“建筑计划”并进行协调，使头部的大脑和运动神经元精确地定位在四肢正在发育的地方。具体来说，这项工作将侧重于开发新工具来研究β-连环蛋白，β-连环蛋白是一种在发育和癌症等疾病中发挥重要作用的分子。由于该分子的重要性，我们将其用作这些新技术的“测试用例”。通过这种方式，我们的“测试案例”将教会我们很多关于这些新方法的知识，并且也可能对未来的生物学研究普遍有用。第二个目标是利用我们现有的糖原合成酶激酶 3 (GSK-3) 的药物依赖性等位基因来研究神经嵴迁移。这种蛋白质也是发育和疾病的重要调节因子。我们之前已经使用类似的方法表明，GSK-3 在味觉发育的关键时期是必需的。使用这些小鼠，我们发现在妊娠中期添加这种蛋白质足以挽救突变小鼠的腭裂。我们希望利用这些方法来了解不同发育过程中所需的基因活动的时间和数量。总而言之，该项目将为研究界提供重要的新化学生物学工具，并深入了解神经嵴迁移的分子机制。更好地了解神经嵴迁移可能有助于我们更好地了解人类发育和癌症转移等疾病过程。

项目成果

Engineering FKBP-Based Destabilizing Domains to Build Sophisticated Protein Regulation Systems.

• DOI: 10.1371/journal.pone.0145783
• 发表时间: 2015
• 期刊: PloS one
• 影响因子: 3.7
• 作者: An W;Jackson RE;Hunter P;Gögel S;van Diepen M;Liu K;Meyer MP;Eickholt BJ
• 通讯作者: Eickholt BJ

Cranial suture lineage and contributions to repair of the mouse skull

• DOI: 10.1242/dev.202116
• 发表时间: 2024-02-01
• 期刊: DEVELOPMENT
• 影响因子: 4.6
• 作者: Doro，Daniel;Liu，Annie;Liu，Karen J.
• 通讯作者: Liu，Karen J.

Calvarial Suture-Derived Stem Cells and Their Contribution to Cranial Bone Repair.

• DOI: 10.3389/fphys.2017.00956
• 发表时间: 2017
• 期刊: Frontiers in physiology
• 影响因子: 4
• 作者: Doro DH;Grigoriadis AE;Liu KJ
• 通讯作者: Liu KJ

Glycogen synthase kinase 3 controls migration of the neural crest lineage in mouse and Xenopus.

• DOI: 10.1038/s41467-018-03512-5
• 发表时间: 2018-03-19
• 期刊: Nature communications
• 影响因子: 16.6
• 作者: Gonzalez Malagon SG;Lopez Muñoz AM;Doro D;Bolger TG;Poon E;Tucker ER;Adel Al-Lami H;Krause M;Phiel CJ;Chesler L;Liu KJ
• 通讯作者: Liu KJ

Cranial neural crest cells form corridors prefiguring sensory neuroblast migration.

• DOI: 10.1242/dev.091033
• 发表时间: 2013-09
• 期刊: Development (Cambridge, England)
• 作者: Freter S;Fleenor SJ;Freter R;Liu KJ;Begbie J
• 通讯作者: Begbie J