Promoting Receptor Protein Tyrosine Phosphatase Activity by Targeting Transmembrane Domain Interactions

通过靶向跨膜结构域相互作用促进受体蛋白酪氨酸磷酸酶活性

• 批准号: 10265510
• 负责人: Matthew J Lazzara
• 金额: $ 39.74万
• 依托单位: LEHIGH UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-20 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10265510
• 关键词: Affect; Agonist; Binding; Biological; Biology; Cancer cell line; Cell Line; Cells; Complement; Computer Models; Data; Development; Dimerization; Disease Progression; Engineering; Epidermal Growth Factor Receptor; Family; Generations; Goals; Health; Heterodimerization; Homo; Homodimerization; Human; Knowledge; Lead; Length; Malignant Neoplasms; Mass Spectrum Analysis; Measurement; Mediating; Membrane; Methods; Modeling; Molecular; Mus; Mutation; Oncogenic; Outcome; PTPRJ gene; Peptides; Phenotype; Phosphoric Monoester Hydrolases; Phosphorylation; Play; Point Mutation; Protein Dephosphorylation; Protein Tyrosine Phosphatase; Receptor Protein-Tyrosine Kinases; Regulation; Reporting; Research; Resistance; Role; Signal Pathway; Signal Transduction; Structure; Structure-Activity Relationship; System; Systems Biology; Tertiary Protein Structure; Testing; Therapeutic; Transmembrane Domain; Trefoil Motif; Tyrosine Kinase Inhibitor; Variant; attenuation; base; cancer cell; cancer type; design; dimer; embryonic stem cell; improved; insight; member; model building; monomer; mutant; novel therapeutic intervention; predictive modeling; prevent; programs; protein expression; receptor; response; therapeutic target; trafficking; tumor; tumor xenograft

PROJECT SUMMARY 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.

项目摘要 受体蛋白酪氨酸磷酸酶（RPTPS）在发育，健康，健康， 和疾病进展。尽管RPTP在信号转导中显然很重要，但鲜为人知 关于基于其活性调节的结构 - 功能关系。报告的能力 RPTP同二聚化以拮抗其催化活性 制定策略来促进对其致癌受体酪氨酸激酶（RTK）底物的活性。 我们最近使用PTPRJ/EGFR作为模型RPTP/RTK对表明：（i）PTPRJ的同构化 （也称为dep1）受跨膜结构域相互作用的调节，（ii）破坏这些 相互作用可以拮抗PTPRJ同二聚化，减少底物EGFR磷酸化，并且 对抗EGFR驱动的细胞表型。 在这里，我们建议沿着三个主题相互联系的这些新见解，但 非重叠的特定目标，其最终目标是：（1）证明RPTP TM域相互作用 对于调节其活动和底物访问至关重要，以及（2）开发一种新的治疗方法 促进对其致癌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异构化的分子决定因素 egfr。这些研究将通过扩展以了解PTPRJ TM域突变体如何补充。 影响受体运输和最终细胞结局。在第二个目标中，我们将设计并选择肽 能够与PTPRJ TM域结合并测试其破坏PTPRJ同二聚化的能力，促进 针对EGFR和其他底物RTK的PTPRJ活性，并选择性地靶向人类肿瘤异种移植物 老鼠。在第三个目标中，我们将确定其他候选RTK基质的ptprj依赖 在TM结构域介导的异二聚化时，并确定不同的细胞环境如何预测细胞 通过TM域干扰PTPRJ二聚化的信号传导和表型结果。为此，我们 将基于表型的数据驱动计算建模来实现系统生物学方法 A蛋白磷酸化和表达的测量和全球质谱测量 细胞系的面板。理解所有RPTP都有多个底物，并且该目标的动机是 当PTPRJ时，这些底物表达的表达变化可能会导致不同的结果 二聚化被破坏。 最终，这里提出的研究旨在提高我们对RPTP的基本生物学理解 生物学，这是非常需要的，并导致通过致癌RTK靶向信号的新方法 可能不太容易受到对RTK抑制剂的耐药性的常见机制。