Promoting Receptor Protein Tyrosine Phosphatase Activity by Targeting Transmembrane Domain Interactions

以下是填写此必填字段的原始项目摘要 受体蛋白酪氨酸磷酸酶(RPTPs)在发育过程中发挥着重要的信号调节作用。 健康和疾病进展。尽管RPTPs在信号转导中具有明显的重要性，但很少有 了解作为其活动调节基础的结构-功能关系。据报道， 然而，RPTP均二聚对抗其催化活性的能力存在潜力 有机会制定策略促进RPTP对其致癌受体酪氨酸激酶的活性 (RTK)基片。最近，我们使用PTPRJ/EGFR作为RPTP/RTK对的模型，表明：(I) PTPRJ(也称为DEP1)的同源二聚受跨膜结构域相互作用的调节，并且 (Ii)破坏这些相互作用可以拮抗PTPRJ同源二聚，减少底物EGFR 磷酸化，并拮抗EGFR驱动的细胞表型。 在这里，我们建议基于这些新的见解，沿着三个主题相互关联但不是- 重叠的、特定的目标，最终目标是：(1)证明RPTP TM域相互作用是 在调节它们的活性和底物获取方面是必不可少的，以及(2)开发一种新的治疗方法 促进针对其致癌RTK底物的RPTP活性。 在我们的第一个目标中，我们将确定调节PTPRJ异二聚化的分子决定因素 EGFR。这些研究将通过扩展它们来理解PTPRJ TM结构域突变是如何 影响受体运输和最终细胞结果。在第二个目标中，我们将设计和选择多肽 能够与PTPRJ TM结构域结合并测试它们破坏PTPRJ同源二聚的能力，促进 PTPRJ对EGFR和其他底物RTK的活性，并选择性靶向人肿瘤移植瘤 老鼠。在第三个目标中，我们将确定其他候选RTK底物，其调控取决于PTPRJ TM结构域介导的异二聚化，并确定不同的细胞环境如何预测细胞 通过TM结构域干扰PTPRJ二聚化的信号和表型结果。为了做到这一点，我们 将实施基于数据驱动的表型计算建模的系统生物学方法 蛋白质磷酸化和表达的测量和全球质谱学测量 细胞系的面板。这一目标的动机是理解所有的RPTP都有多个底物，并且 这些底物在细胞间表达的差异可能会导致PTPRJ时不同的结果 二聚化被打乱了。 最终，这里提出的研究将促进我们对RPTP的基本生物学理解 生物学，这是迫切需要的，并导致通过致癌RTK靶向信号的新方法， 可能对常见的RTK抑制剂获得性耐药机制不太敏感。

Below is the original Project Summary to fill this mandatory field Receptor protein tyrosine phosphatases (RPTPs) play critical signaling regulatory roles in development, health, and disease progression. Despite the clear importance of RPTPs in signal transduction, very little is known about the structure-function relationships that underpin the regulation of their activity. The reported ability of RPTP homodimerization to antagonize their catalytic activity, however, presents potential opportunities to develop strategies to promote RPTP activity against their oncogenic receptor tyrosine kinase (RTK) substrates. We recently showed, using PTPRJ/EGFR as a model RPTP/RTK pair, that: (i) homodimerization of PTPRJ (also known as DEP1) is regulated by transmembrane domain interactions, and (ii) disrupting these interactions can antagonize PTPRJ homodimerization, reduce substrate EGFR phosphorylation, and antagonize EGFR-driven cell phenotypes. Here, we propose to build upon these new insights along three thematically interconnected, but non- overlapping, specific aims, with the ultimate goals of: (1) demonstrating that RPTP TM domain interactions are essential in regulating their activity and substrate access, and (2) developing a new therapeutic approach to promote RPTP activity against their oncogenic RTK substrates. In our first aim, we will determine the molecular determinants regulating the heterodimerization of PTPRJ with EGFR. These studies will be complemented by extending them to understand how PTPRJ TM domain mutants affect receptor trafficking and ultimate cell outcomes. In the second aim, we will design and select peptides capable of binding to PTPRJ TM domains and test their ability to disrupt PTPRJ homodimerization, promote PTPRJ activity against EGFR and other substrate RTKs, and selectively target human tumor xenografts in mice. In the third aim, we will identify other candidate RTK substrates whose regulation by PTPRJ depends upon TM domain-mediated heterodimerization, and determine how different cellular contexts predict the cell signaling and phenotype outcome of interfering with PTPRJ dimerization through TM domains. To do so, we will implement a systems biology approach based on data-driven computational modeling of phenotypic measurements and global mass spectrometry measurements of protein phosphorylation and expression in a panel of cell lines. This aim is motivated by an understanding that all RPTPs have multiple substrates and that variations in expression of those substrates among cells may lead to different outcomes when PTPRJ dimerization is disrupted. Ultimately, the studies proposed here stand to advance both our basic biological understanding of RPTP biology, which is critically needed, and to lead to new methods to target signaling through oncogenic RTKs that may be less susceptible to common mechanisms of acquired resistance to RTK inhibitors.