Targeting PLK1 signaling for the treatment of fibrolamellar carcinoma

靶向 PLK1 信号传导治疗纤维板层癌

• 批准号: 10742683
• 负责人: Taran Singh Gujral
• 金额: $ 49.23万
• 依托单位: FRED HUTCHINSON CANCER CENTER
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-01 至 2028-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10742683
• 关键词: Acceleration; Address; Adolescent and Young Adult; Affect; Binding; Biochemical; Biochemistry; Biological Assay; Biological Models; Biological Process; CRISPR/Cas technology; Cancer Model; Catalytic Domain; Cell Cycle; Cell Line; Cell Proliferation; Cell Survival; Cell division; Cells; Centrosome; Chemoresistance; Childhood Liver Cancer; Chimera organism; Chimeric Proteins; Clinical; Clinical Trials; Co-Immunoprecipitations; Collaborations; Colorectal Cancer; Complex; Cyclic AMP-Dependent Protein Kinases; Data; Development; Disease; Disease Resistance; Event; Excision; Expression Profiling; Fibrolamellar Hepatocellular Carcinoma; Gene Fusion; Generations; Genetic; Goals; Gravin; Growth; Hepatocyte; Histology; Human; Immunofluorescence Immunologic; In Vitro; Knowledge; Life; Ligase; Liver; Machine Learning; Malignant Epithelial Cell; Malignant neoplasm of liver; Mitosis; Mitotic; Modeling; Molecular; Mus; N-terminal; Operative Surgical Procedures; Oral; PLK1 gene; Pathogenesis; Patients; Pharmacology; Pharmacotherapy; Phenotype; Phosphotransferases; Pre-Clinical Model; Property; Protein Array; Proteins; RNA Interference; Research Personnel; Resistance; Role; Signal Transduction; Slice; System; Systems Biology; Testing; Therapeutic; Tissues; Tumor Weights; Up-Regulation; Variant; Work; advanced disease; candidate validation; cell growth; chemotherapy; clinical application; clinical candidate; clinical efficacy; druggable target; efficacy evaluation; experimental study; follow-up; genetic signature; heat-shock proteins 40; inhibitor; insight; irinotecan; kinase inhibitor; knock-down; mortality; novel; novel therapeutic intervention; novel therapeutics; patient derived xenograft model; pharmacologic; polo-like kinase kinase 1; preclinical trial; research clinical testing; response; screening; stem; targeted treatment; trait; tumor; tumor growth; tumor progression

PROJECT SUMMARY Fibrolamellar carcinoma (FLC) is a childhood liver cancer with a high case mortality rate. Patients with FLC typically present with advanced disease, as there are no early warning signs. Thus, a cure by surgical resection is rarely attainable. In addition, FLCs are notoriously resistant to chemotherapies and other targeted therapies currently approved for liver cancer, leading to a 5-year survival of just 30%. New therapeutic strategies that counteract the molecular signaling events that go awry in FLC are urgently needed. FLC is characterized by a fusion event resulting in a novel chimeric protein that joins the N-terminal domain of DNAJ with the catalytic subunit of protein kinase A (PKAc) in hepatocytes. However, the underlying mechanism by which DNAJ-PKAc drives FLC tumor growth remains unknown. This project's overall goal is to apply an unbiased systems-based approach to identify and validate druggable signaling networks that regulate the growth of DNAJ-PKAc- expressing FLC cells and uncover a mechanistic understanding of how DNAJ-PKAc chimeric protein drives FLC. The paucity of preclinical models such as immortalized primary human FLC cell lines has precluded many investigators. Our lab has established three new model systems to address this significant gap: patient-derived cell lines bearing the FLC gene fusion, organotypic cultures, and patient-derived xenograft (PDX) mice. Utilizing these model systems, we carried out a systems-pharmacology-based functional kinase inhibitor screening in FLC cells and normal hepatocytes. We identified and confirmed the role of PLK1 kinases as essential for the growth of FLC cells. Genetic depletion or pharmacological inhibition of PLK1 selectively reduces the growth of multiple patient-derived FLC cell lines and the viability of FLC organotypic tissue slices. Further, treatment of the FLC tumor with PLK1 inhibitor significantly reduced the tumor growth in the PDX model. PLK kinases are key regulators of centrosome maturation and mitosis. Follow-up experiments suggest that DNAJ-PKAc chimera localizes to the centrosomes where it physically interacts with PLK1.Thus, we hypothesize that the heightened sensitivity of the FLC cells to PLK inhibition stems from the localization of the DNAJ-PKAc fusion protein to the centrosome, its association with the PLK1 complex, thereby enhancing the activation of PLK1 and promoting mitotic progression. We propose to 1. uncover molecular mechanisms of how DNAJ-PKAc fusion alters PLK1 activation and function, and 2. evaluate the efficacy of clinical-grade PLK1 inhibitors alone and in combination with chemotherapy in preclinical models.Functional analyses will highlight the mechanistic insights by which DNAJ-PKAc drives FLC tumor progression and the role of the PLK1 signaling complex in FLC survival, thus deepening our understanding of disease pathogenesis. Our cross-disciplinary team consisting of Drs. Gujral, Scott, and Yeung represent a cohesive collaboration that brings systems biology, PKA biochemistry, and state- of-art human-derived FLC cancer models to address this deadly disease. Our findings have translational significance as they will provide a rationale for targeting critical signaling nodes that sustain FLC tumors' survival.

项目摘要 纤维素癌（FLC）是一种儿童肝癌，病例死亡率很高。 FLC患者 通常存在晚期疾病，因为没有预警信号。因此，通过手术切除治疗 很少可以实现。此外，众所周知，FLC对化学疗法和其他靶向疗法具有抗性 目前已批准肝癌，导致5年生存仅为30％。新的治疗策略 迫切需要抵消在FLC中出现问题的分子信号传导事件。 FLC的特征是 融合事件导致一种新型的嵌合蛋白，该嵌合蛋白与DNAJ的N末端结构域与催化 肝细胞中蛋白激酶A（PKAC）的亚基。但是，DNAJ-PKAC的基本机制 驱动FLC肿瘤生长仍然未知。 该项目的总体目标是应用基于系统的公正 识别和验证可吸毒信号网络的方法，以调节DNAJ-PKAC-的生长 表达FLC细胞并发现对DNAJ-PKAC嵌合蛋白如何驱动FLC的机械理解。 临床前模型（例如永生的原发性人类FLC细胞系）的匮乏排除了许多 调查人员。我们的实验室已经建立了三个新的模型系统来解决这一重大差距：患者衍生 带有FLC基因融合，器官型培养物和患者衍生异种移植（PDX）小鼠的细胞系。利用 这些模型系统，我们进行了基于系统 - 药理学的功能激酶抑制剂筛选 FLC细胞和正常的肝细胞。我们确定并确认了PLK1激酶对 FLC细胞的生长。 PLK1的遗传耗竭或药理抑制选择性降低 多个患者衍生的FLC细胞系和FLC器官组织切片的生存能力。此外，处理 具有PLK1抑制剂的FLC肿瘤可显着降低PDX模型的肿瘤生长。 PLK激酶是关键 中心体成熟和有丝分裂的调节剂。后续实验表明DNAJ-PKAC嵌合体 定位于与PLK1物理相互作用的中心体。 FLC细胞对PLK抑制的敏感性源于DNAJ-PKAC融合蛋白的定位 中心体，与PLK1复合物的关联，从而增强了PLK1的激活并促进 有丝分裂进展。我们提议1。发现DNAJ-PKAC融合如何改变PLK1的分子机制 激活和功能，以及2。单独和组合评估临床级PLK1抑制剂的功效 通过临床前模型中的化学疗法。功能分析将突出显示的机械见解 DNAJ-PKAC驱动FLC肿瘤进展和PLK1信号复合物在FLC存活中的作用，因此 加深我们对疾病发病机理的理解。我们的跨学科团队由DRS组成。古杰拉尔， 斯科特（Scott）和杨代表着一个有凝聚力的合作，它带来了系统生物学，PKA生物化学和国家 - ART人类衍生的FLC癌症模型解决了这种致命的疾病。我们的发现有翻译 意义，因为它们将为靶向维持FLC肿瘤生存的关键信号节点的基本原理。