Investigating degradation as a therapeutic strategy against the Pseudo-kinase KSR

项目总结 我的博士研究经验主要集中在使用化学工具来纠正 生物系统，主要是癌症。工作人员实验室中最近开发的一种工具--蛋白质分解 靶向嵌合体(PROTACs)，允许在没有任何 基因操纵。到目前为止，我的重点是将PROTAC作为一个有价值的工具来建立。我 已经表明这些化合物的行为符合我们提出的机制，即它们 特定和有效地降解感兴趣的蛋白质，并且PROTAC可以降解它们的 动物模型中的靶标。在我的F99阶段，我将专注于开发选择性配体 可用于PROTAC以降解假性激酶KSR。这种蛋白质没有突变或 在癌症中被激活，但上调Ras-MAPK通路，参与许多不同的癌症。 通过将高通量筛选与药物化学相结合，我将首先开发 已知的激酶抑制剂库中的选择性配体。一旦配体被开发出来，一个 PROTAC将在该配体的基础上合成，并评估KSR在细胞中的降解情况。我 然后，我将使用此工具进一步阐明KSR在RAS-MAPK中的脚手架角色 途径和其他细胞信号通路通过使用高含量的多肽阵列。这个项目 是将化学生物学工具与癌症的“系统层面”洞察相结合的一个例子。在我的 K00阶段，我希望通过接受高内容剖析方面的培训来延续这种思路 用于研究经常被改变和错误调节的复杂生化环境的技术 并确定新的靶点。通过将整合的“组学”技术与化学 生物学方面，我将致力于为新的靶标开发新的化学工具。

PROJECT SUMMARY My doctoral research experience has focused on using chemical tools to correct perturbed biological systems, primarily cancer. A recently develop tool in the Crews’ lab, proteolysis targeting chimera (PROTACs), allows for the degradation of a protein of interest without any genetic manipulation. Thus far, I have focused on establishing PROTACs as a valuable tool. I have shown that these compounds behave according to our proposed mechanism, that they specifically and potently degrade the protein of interest, and that PROTACs can degrade their targets in animal models. For my F99 phase, I will focus on developing selective ligands that can be used in a PROTAC to degrade the pseudo-kinase KSR. This protein is not mutated or activated in cancer, but upregulates the Ras-MAPK pathway involved in many different cancers. By combining high-throughput screening with medicinal chemistry, I will first develop the selective ligand from a library of known kinase inhibitors. Once a ligand is developed, a PROTAC will be synthesized based on that ligand and assessed for KSR-degradation in cells. I will then use this tool to further clarify the scaffolding roles of KSR in the Ras-MAPK pathway and other cellular signaling pathways by using high-content peptide arrays. This project is an example of combining chemical biology tools with ‘systems-level’ insights in cancer. In my K00 phase, I hope to continue this line of thinking by being trained in high-content profiling techniques used to study the complex biochemical milieu that is often altered and mis-regulated in cancer and identify new targets. By combining integrative ‘-omics’ techniques with chemical biology, I will work to develop novel chemical tools for novel targets.