Cyclin C-CDK8/19 kinases in development and in cancer

发育和癌症中的细胞周期蛋白 C-CDK8/19 激酶

基本信息

批准号：
10579308
负责人：
Peter Sicinski
金额：
$ 51.55万
依托单位：
DANA-FARBER CANCER INST
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-03-01 至 2027-02-28
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10579308
关键词：
Ablation Address Affect Alanine Allografting Alpha Interleukin 2 Receptor Amino Acids Animals Antitumor Response Binding Biochemical CD8-Positive T-Lymphocytes CD8B1 gene CLN1 gene CLN2 gene CLN3 gene Cancer Model Cancer Patient Catalytic Domain Cell Culture Techniques Cell Lineage Cells Clinical Clinical Trials Combined Modality Therapy Complementary DNA Complex Cultured Cells Cyclin-Dependent Kinases Cyclins Cytotoxic T-Lymphocytes Development Endothelial Cells Enterobacteria phage P1 Cre recombinase Gatekeeping Genetic Transcription Growth Half-Life Human IL2RA gene IL2RB gene Immune Immune response Immune system Immunocompromised Host In Vitro Interleukin 2 Receptor Interleukin-2 Interleukin-4 Knockout Mice Laboratories Link LoxP-flanked allele Lymphoid Cell Malignant Neoplasms Maps Mediating Mediator Memory Modality Molecular Mus Mutation NKTR gene Oncogenes Outcome Patients Peripheral Phosphorylation Phosphotransferases Play Polyubiquitination Protein Complex Subunit Proteins RNA Polymerase II Regulatory T-Lymphocyte Research Role Saccharomyces cerevisiae Signal Transduction Site T cell differentiation T-Cell Development T-Lymphocyte T-Lymphocyte Subsets Testing Thymus Gland Transcript Tumor Immunity Tumor Promotion Up-Regulation Wild Type Mouse Work Xenograft procedure Yeasts ZNF145 gene adaptive immune response adaptive immunity anti-CTLA4 antibodies anti-PD-1 anti-cancer anti-tumor immune response antitumor effect cancer cell cancer therapy cyclin C cyclin G1 effector T cell experimental study genetic approach immune checkpoint blockade in vivo inhibitor mouse genetics novel overexpression paralogous gene peripheral lymphoid organ small molecule small molecule inhibitor systemic toxicity therapeutic target thymocyte transcription factor tumor tumorigenesis ubiquitin-protein ligase

项目摘要

Project Summary/Abstract Cyclin C was cloned, along with other G1 cyclins, as a cDNA which can rescue the proliferative block in yeast Saccharomyces cerevisiae lacking CLN1, CLN2 and CLN3 genes. Subsequent work revealed that cyclin C, acting together with its kinase partners, the cyclin-dependent kinase 8 (CDK8) and CDK19, plays a role in regulating gene transcription. Cyclin C serves as a regulatory partner of CDK8 and CDK19, and activates the kinase activity of these proteins. The two paralog kinases show substantial overall amino acid identity, particularly within their catalytic domains. Cyclin C, CDK8 and CDK19 represent parts of the mediator complex, a large multisubunit protein complex that regulates gene transcription by linking RNA polymerase II to sequence-specific transcription factors. In addition, cyclin C-CDK8/19 was shown to phosphorylate various transcription factors and to regulate their stability and activity. Growing evidence indicates that cyclin C-CDK8 and C-CDK19 kinases may represent potential anti-cancer targets. CDK8 has been identified as an oncogene in several human cancers. Cyclin C, CDK8 and CDK19 are overexpressed in a wide range of tumor types. Importantly, higher expression of these three proteins was found to be associated with poor clinical outcome. Consistent with tumor-promoting roles for cyclin C-CDK8/19, treatment of mice bearing xenografts of several human tumor types with small molecule CDK8/19 inhibitors resulted in a potent anti-tumor effect without apparent systemic toxicity. To understand the molecular function of cyclin C in vivo, we generated conditional cyclin C knockout mice (cyclin CF/F). To test the impact of cyclin C-CDK8/19 inhibition on T cell development and adaptive anti-tumor immunity, we crossed cyclin CF/F mice with CD4-Cre animals. The latter strain expresses Cre recombinase at an early stage of T cell development, and drives deletion of the ‘floxed’ sequences in all T cell subsets. In our preliminary analyses we established that cyclin C functions as a gate- keeper of T cell differentiation, likely by directly phosphorylating a lineage-specific transcription factor. By doing so, cyclin C affects signaling networks that regulate the immune response. Hence, this novel function of cyclin C may have a profound effect on the anti-tumor immune response and on tumorigenesis. In the proposed work we will extend these studies. In Aim 1, we will study the exact molecular function of cyclin C- CDK8 and C-CDK19 in the T cell lineage using a combination of biochemical in vitro studies, cell culture experiments, analyses of ex vivo cultured T cells as well as mouse genetic approaches. In Aim 2, we will study how ablation of cyclin C impacts tumorigenesis in vivo, using mouse cancer models and combination treatments with other anti-cancer modalities. In Aim 3, we will extend these cancer studies to cyclin C catalytic partners, using conditional Cdk19 knockout mice (Cdk19F/F), that my laboratory generated for this purpose. The expected overall impact of this proposal is that it will reveal a novel and important function of cyclin C- CDK8 and C-CDK19 in tumorigenesis, and will validate these kinases as attractive therapeutic targets.

项目概要/摘要细胞周期蛋白 C 与其他 G1 细胞周期蛋白一起被克隆为 cDNA，可以挽救酵母中的增殖阻断缺乏 CLN1、CLN2 和 CLN3 基因的酿酒酵母随后的工作表明，细胞周期蛋白 C，与其激酶伙伴、细胞周期蛋白依赖性激酶 8 (CDK8) 和 CDK19 一起作用，在 Cyclin C 是 CDK8 和 CDK19 的调节伙伴，并激活这些蛋白质的激酶活性显示出显着的整体氨基酸同一性，特别是在它们的催化域内，CDK8 和 CDK19 代表介体的一部分。复合物，一种大型多亚基蛋白质复合物，通过将 RNA 聚合酶 II 连接到来调节基因转录此外，细胞周期蛋白 C-CDK8/19 还可磷酸化多种转录因子。越来越多的证据表明细胞周期蛋白 C-CDK8 可以调节转录因子的稳定性和活性。 C-CDK19 激酶可能代表潜在的抗癌靶标，CDK8 已被确定为癌基因。在多种人类癌症中，Cyclin C、CDK8 和 CDK19 在多种肿瘤类型中过度表达。重要的是，这三种蛋白的较高表达被发现与不良的临床结果相关。与细胞周期蛋白 C-CDK8/19 的肿瘤促进作用一致，对携带几种异种移植物的小鼠进行治疗小分子 CDK8/19 抑制剂对人类肿瘤类型产生了有效的抗肿瘤作用，且无需为了了解细胞周期蛋白 C 的体内分子功能，我们生成了条件条件。细胞周期蛋白 C 敲除小鼠 (细胞周期蛋白 CF/F) 测试细胞周期蛋白 C-CDK8/19 抑制对 T 细胞发育的影响。和适应性抗肿瘤免疫，我们将细胞周期蛋白 CF/F 小鼠与 CD4-Cre 动物杂交。在 T 细胞发育的早期阶段表达 Cre 重组酶，并驱动“floxed”的删除在我们的初步分析中，我们确定细胞周期蛋白 C 具有门控功能。 T 细胞分化的保持者，可能是通过直接磷酸化谱系特异性转录因子来实现的。这样做，细胞周期蛋白 C 会影响调节免疫反应的信号网络，因此产生了这一新功能。细胞周期蛋白 C 可能对抗肿瘤免疫反应和肿瘤发生具有深远的影响。在目标 1 中，我们将研究细胞周期蛋白 C- 的确切分子功能。结合体外生化研究、细胞培养研究 T 细胞谱系中的 CDK8 和 C-CDK19 在目标 2 中，我们将研究实验、离体培养 T 细胞分析以及小鼠遗传方法。使用小鼠癌症模型和组合，消除细胞周期蛋白 C 如何影响体内肿瘤发生在目标 3 中，我们将这些癌症研究扩展到细胞周期蛋白 C 催化。合作伙伴，使用我的实验室为此目的培育的条件性 Cdk19 敲除小鼠 (Cdk19F/F)。该提案的预期总体影响是，它将揭示细胞周期蛋白 C 的一个新颖且重要的功能—— CDK8 和 C-CDK19 在肿瘤发生中的作用，并将验证这些激酶作为有吸引力的治疗靶点。