Stabilizing nuclear p27kip1 as a therapeutic target for endometrial cancer

稳定核 p27kip1 作为子宫内膜癌的治疗靶点

基本信息

批准号：
8698061
负责人：
TIMOTHY J CARDOZO
金额：
$ 63.97万
依托单位：
NEW YORK UNIVERSITY SCHOOL OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-06-13 至 2019-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8698061
关键词：
Address Affect Atypical Endometrial Hyperplasias Binding Biological Availability Biological Markers Biopsy Cancer Biology Cell Cycle Cell Cycle Proteins Cell Cycle Regulation Cell Nucleus Cell Proliferation Cells Cessation of life Chemicals Complex Crystallization Cyclin-Dependent Kinase Inhibitor Data Development Disease Endometrial Endometrial Carcinoma Endometrium Epithelial Epithelial Cell Proliferation Epithelial Cells Estrogens Event G1 Arrest G1 Phase Generations Growth Growth Inhibitors Gynecologic Hormonal Carcinogenesis Hormones Human Image Analysis In Vitro Injection of therapeutic agent Knowledge Lead Learning Left Libraries Malignant - descriptor Malignant Neoplasms Mediating Microscopy Molecular Molecular Target Mus Neoplasm Metastasis Nuclear Oncogenes Outcome Pathogenesis Pathway interactions Patients Pharmaceutical Preparations Progesterone Proteasome Inhibitor Proteins Publishing Regulation Reporting Role System Testing Therapeutic Therapeutic Agents Therapeutic Intervention Therapeutic Uses Tissues Toxic effect Tumor Suppressor Proteins Tumor Volume Ubiquitin X-Ray Crystallography Xenograft procedure cancer cell cancer therapy carcinogenesis cellular imaging chemical group in vivo inhibitor/antagonist insight knock-down migration mouse model multicatalytic endopeptidase complex new therapeutic target novel novel strategies novel therapeutic intervention pharmacophore prevent protein degradation public health relevance scaffold small molecule spatiotemporal therapeutic target trafficking treatment strategy tumor ubiquitin ligase ubiquitin-protein ligase

项目摘要

DESCRIPTION (provided by applicant): As loss of cell cycle regulation is a hallmark of carcinogenesis, identifying novel therapeutic targets aimed at restoring growth control is an important approach to cancer therapy. Endometrial cancer (ECA) is the most common gynecologic malignancy in the US with 49,470 new cases and over 8,190 deaths estimated for 2013. The major objectives of this proposal address mechanisms involved in the pathogenesis of estrogen (E2)- induced Type I ECA (85% of ECAs) and a new potential therapy for this disease that involves inhibiting degradation of the tumor suppressor, p27kip1 (p27), a key cell cycle protein that arrests cell proliferation. We reported that E2 induces ubiquitin-mediated degradation of nuclear p27 by its specific E3 ligase, SCF- Skp2/Cks1, in endometrial epithelial (EECs) and ECA cells with increased cell proliferation. Conversely, progesterone (Pg; a therapy for low grade ECA) as well as TGF-?, increase p27 in the nucleus and inhibit cell proliferation by preventing p27 degradation. Therefore, we provide compelling evidence that p27 is a key target for growth regulation in the endometrium and in endometrial carcinogenesis. We have identified novel small molecule inhibitors of Skp2/Cks1 (Skp2E3LIs) that only increase nuclear p27, which is critical since cytoplasmic p27 mediated migration/metastasis. Furthermore, Skp2E3LIs block both E2-induced degradation of nuclear p27 and proliferation in vitro in ECA cells and {in vivo in mouse EECs}. Importantly, Skp2E3LIs have the potential to be a major therapeutic advancement over current general proteasome inhibitors that indiscriminately block protein degradation including oncogenes. Whereas Skp2/Cks1 causes p27 degradation, the E3 ligase APC/Cdh1 targets Cks1/Skp2 for destruction leaving p27 intact. Interestingly, we show that E2 decreases Cdh1and thereby increases Skp2/Cks1 for p27 degradation whereas Pg and TGF-?? increase Cdh1 to increase nuclear p27. Therefore, p27 degradation can be inhibited both, by increasing Cdh1 or by blocking Skp2. As such, we will test two main hypotheses: 1. that the Cdh1-Skp2/Cks1-p27axis is important in cell cycle dysregulation by the ubiquitin proteasome system (UPS); 2. that Skp2E3LIs have significant translational value for the treatment of ECA. We propose to: 1. Use E2, Pg, and TGF-?? as molecular switches to learn how Cdh1 is controlled to affect p27 levels. 2. Show that co-localization of p27 and Cdh1 in human biopsy tissue is a biomarker for positive outcomes of Pg therapy. {3. Perform co-crystallization/NMR structural analysis of Skp2-Cks1-Skp2E3LI complexes for chemically optimizing current lead compounds to define the role of p27 in ECA in vitro and in vivo.} 4. Quantify the effects of Skp2E3LIs on intracellular [nuclear] trafficking of p27 Skp2, and Cks1 together with cell cycle analysis by single cell imaging microscopy. 5. Test Skp2E3LIs for their efficacy in blocking the growth of human ECA tumors in mouse models. As the degradation of nuclear p27 occurs in numerous cancers, our studies should impact the cancer biology field by providing mechanistic insights into the role of the UPS in cancer and the use of Skp2E3LIs as a novel approach to cancer therapy.

描述（由申请人提供）：由于细胞周期调节的丧失是致癌的标志，因此识别旨在恢复生长控制的新型治疗靶标是癌症治疗的重要方法。 Endometrial cancer (ECA) is the most common gynecologic malignancy in the US with 49,470 new cases and over 8,190 deaths estimated for 2013. The major objectives of this proposal address mechanisms involved in the pathogenesis of estrogen (E2)- induced Type I ECA (85% of ECAs) and a new potential therapy for this disease that involves inhibiting degradation of the tumor suppressor, P27KIP1（P27），一种钥匙细胞周期蛋白，可阻止细胞增殖。我们报道了E2在子宫内膜上皮（EEC）和ECA细胞中诱导其特异性E3连接酶SCF-SKP2/CKS1诱导泛素介导的核P27降解，并随细胞增殖增加。相反，孕酮（PG；低级ECA的治疗）以及TGF-？，增加细胞核中的P27并通过防止P27降解来抑制细胞增殖。因此，我们提供了令人信服的证据，表明p27是子宫内膜和子宫内膜癌发生中生长调节的关键目标。我们已经确定了仅增加核P27的SKP2/CKS1（SKP2E3LIS）的新型小分子抑制剂，这是至关重要的，因为细胞质P27介导的迁移/转移。此外，SKP2E3LIS阻止了E2诱导的ECA细胞中E2诱导的核P27降解和体外增殖的降解和{小鼠EECS中的体内}。重要的是，SKP2E3LI有可能成为当前一般蛋白酶体抑制剂的主要治疗进步，该蛋白酶体抑制剂不加选择地阻止蛋白质降解，包括癌基因。而SKP2/CKS1导致p27降解，而E3连接酶APC/CDH1的目标是CKS1/SKP2破坏，使P27完整。有趣的是，我们表明E2降低了CDH1，从而增加了P27降解的SKP2/CKS1，而PG和TGF-？增加CDH1以增加核P27。因此，可以通过增加CDH1或阻止SKP2来抑制P27降解。因此，我们将检验两个主要的假设：1。CDH1-SKP2/CKS1-P27AXI在泛素蛋白酶体系统（UPS）中对细胞周期失调很重要。 2。该SKP2E3LI在ECA的处理中具有显着的翻译价值。我们建议：1。使用E2，PG和TGF- ??随着分子开关以了解如何控制CDH1以影响P27水平。 2。证明P27和CDH1在人体活检组织中的共定位是PG治疗阳性结果的生物标志物。 {3。对SKP2-CKS1-SKP2E3LI复合物进行联合结晶/NMR结构分析，以化学优化电流铅化合物，以定义p27在ECA体外和体内的作用。} 4。量化SKP2E3LIS对P27 SKP2和CCKS的细胞内[核]型单元格的影响。 5。测试SKP2E3LI在阻断小鼠模型中人类ECA肿瘤生长方面的功效。随着核p27的降解发生在许多癌症中，我们的研究应通过提供机械洞察力来影响癌症生物学领域，以了解UPS在癌症中的作用以及使用SKP2E3LIS作为癌症治疗的新方法。