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Novel regulation of the CDK4/Cdh1/Pin1 signaling axis for targeted breast cancer therapies

用于乳腺癌靶向治疗的 CDK4/Cdh1/Pin1 信号轴的新调控

基本信息

批准号：
10281417
负责人：
Fabin Dang
金额：
$ 16.99万
依托单位：
BETH ISRAEL DEACONESS MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-07-01 至 2023-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10281417
关键词：
Adaptor Signaling Protein Anaphase Breast Cancer Cell Breast Cancer therapy CDK4 gene Cell Culture Techniques Cell Cycle Cell Cycle Arrest Cell Cycle Progression Cell Cycle Proteins Cell Cycle Regulation Cell Proliferation Cells Clinical Complex Couples Cyclin D1 Data Development Employment Exhibits G1 Phase Genetic Goals Human Immune In Vitro Maintenance Malignant Neoplasms Mediating Metaphase Mitosis Modeling Molecular Mutate Oncogenic Oncoproteins Pathway interactions Peptidylprolyl Isomerase Phosphorylation Phosphorylation Site Phosphotransferases Physiological Pin1 protein Play Polyubiquitination Process Proteins RB1 gene Regulation Role S Phase Serine Signal Transduction Therapeutic Therapeutic Effect Threonine Tumor Suppressor Proteins Ubiquitin Work anaphase-promoting complex breast tumorigenesis cancer stem cell cancer therapy design in vivo inhibitor/antagonist kinase inhibitor malignant breast neoplasm mouse genetics multicatalytic endopeptidase complex novel novel therapeutics overexpression prevent protein degradation protein expression protein function response tumorigenesis ubiquitin-protein ligase

项目摘要

Project Summary/Abstract: The anaphase-promoting complex/Cyclosome (APC/C) is a well-defined multi-subunit E3 ubiquitin ligase that regulates targeted cell cycle regulators for degradation by the Ubiquitin Proteasome Pathway (UPP), promoting cell cycle progression from metaphase to anaphase and being involved in G1 phase maintenance. The APC/C E3 ligase complex is evolutionarily conserved and relies on two adaptor proteins, Cdc20 and Cdh1, to recognize different target proteins and regulate cell cycle progression. However, compared to Cdc20 that is subjected to Cdh1-mediated destruction, regulation of the E3 ligase activity of Cdh1 is not well known yet. Previous study has shown that there were 19 serine and threonine residues on Cdh1 that can be phosphorylated by multi-kinases in vivo, indicating that the phosphoregulation of Cdh1 is much more complex. In the present proposal, I found that CDK4 can phosphorylates Cdh1 in vitro and modulates its E3 ligase activity. Furthermore, we found that the phosphorylation of Cdh1 by CDK4 can be recognized by the Pin1 proline isomerase, facilitating Cdh1-Pin1 complex formation. In keeping with this notion, employment of the CDK4/6 inhibitor or mutating the phosphorylation sites can disrupt the Cdh1-Pin1 interaction. Consequently, Cdh1 can mediate Pin1 for polyubiquitination and degradation. As such, depletion of endogenous Cdh1 abolished the Pin1 inhibitor treatment induced Pin1 degradation in cells. Importantly, the Pin1 inhibitor-induced cell proliferation suppression was also abolished in Cdh1-null MEFs, suggesting the functional presence of Cdh1 is required for Pin1 inhibitor-induced cell proliferation suppression. In addition, combination of the CDK4/6 inhibitor and Pin1 inhibitor exhibits significantly enhanced suppressing effect in breast cancer cells. In the first Aim of this proposal, I am going to explore the role of CDK4 kinase in regulating the E3 ligase activity of Cdh1 (Aim #1). Therefore, the second Aim in this proposal will be exploring the potential role of Cdh1 in mediating Pin1 inhibitor treatment induced Pin1 protein destruction (Aim 2). Together, these results implicate a functional role of the CDK4/Cdh1/Pin1 signaling axis in regulating cell proliferation, and provide rational for combining the CDK4/6 inhibitor and Pin1 inhibitor to treat breast cancer.

项目摘要/摘要：后期促进复合物/环体 (APC/C) 是一种明确的多亚基 E3 泛素调节目标细胞周期调节剂通过泛素蛋白酶体途径降解的连接酶 (UPP)，促进细胞周期从中期到后期的进展并参与G1期维护。 APC/C E3 连接酶复合物在进化上是保守的，并且依赖于两个接头蛋白， Cdc20 和 Cdh1，识别不同的靶蛋白并调节细胞周期进程。然而，与受到 Cdh1 介导的破坏的 Cdc20 相比，E3 连接酶活性的调节 Cdh1 尚不为人所知。此前的研究表明，其上有19个丝氨酸和苏氨酸残基。 Cdh1在体内可被多种激酶磷酸化，表明Cdh1的磷酸调节作用是复杂得多。在本提案中，我发现CDK4可以在体外磷酸化Cdh1，并且调节其 E3 连接酶活性。此外，我们发现CDK4对Cdh1的磷酸化可以被 Pin1 脯氨酸异构酶识别，促进 Cdh1-Pin1 复合物的形成。与此相一致概念，使用 CDK4/6 抑制剂或突变磷酸化位点可以破坏 Cdh1-Pin1 相互作用。因此，Cdh1 可以介导 Pin1 的多泛素化和降解。如此一来，耗尽内源性 Cdh1 的消除消除了 Pin1 抑制剂处理诱导的细胞中 Pin1 降解。重要的是， Pin1 抑制剂诱导的细胞增殖抑制在 Cdh1 缺失的 MEF 中也被消除，这表明 Pin1 抑制剂诱导的细胞增殖抑制需要 Cdh1 的功能性存在。在此外，CDK4/6 抑制剂和 Pin1 抑制剂的组合表现出显着增强的抑制作用对乳腺癌细胞的作用。在本提案的第一个目标中，我将探讨 CDK4 激酶在调节 Cdh1 的 E3 连接酶活性（目标#1）。因此，本提案的第二个目标是探索 Cdh1 在介导 Pin1 抑制剂治疗诱导的 Pin1 蛋白破坏中的潜在作用（目标 2）。总之，这些结果表明 CDK4/Cdh1/Pin1 信号轴在调节中的功能作用细胞增殖，为CDK4/6抑制剂与Pin1抑制剂联合治疗乳腺提供理论依据癌症。