Combined inhibition of PLK1 and NOTCH for melanoma management

联合抑制 PLK1 和 NOTCH 治疗黑色素瘤

• 批准号: 10481129
• 负责人: Nihal Ahmad
• 金额: --
• 依托单位: WM S. MIDDLETON MEMORIAL VETERANS HOSP
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-01-01 至 2026-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10481129
• 关键词: ABCB1 gene; ABCC1 gene; ANXA5 gene; Affect; BRAF gene; Biological; Blood Vessels; Cell Cycle; Cell Cycle Regulation; Cell Line; Cell Proliferation; Cells; Characteristics; Climate; Clinical; Clinical Management; Combined Modality Therapy; Data; Development; Diagnosis; Disease; Disease Progression; Drug resistance; Embryo; Epithelium; Evolution; Extracellular Matrix; FDA approved; Family member; Gene Expression Profiling; Genes; Genetic Engineering; Goals; Growth; Healthcare; Human; Immune checkpoint inhibitor; Immunotherapy; Laboratories; Link; MEKs; Malignant Neoplasms; Melanoma Cell; Mesenchymal; Metastatic Melanoma; Military Personnel; Mission; Mitosis; Modeling; Mutation; NOTCH1 gene; Neoplasm Metastasis; Neoplasms; Oncogenic; Outcome; PLK1 gene; Pathway interactions; Patients; Pharmaceutical Preparations; Phosphopeptides; Play; Preclinical Testing; Progression-Free Survivals; Proliferating; Protein-Serine-Threonine Kinases; Proteins; Proteome; Proteomics; Publishing; Relapse; Reporting; Research; Resistance; Resistance development; Role; Sampling; Serine; Signal Transduction; Techniques; Testing; Therapeutic; Tissues; Treatment Efficacy; Treatment outcome; Tumor Markers; United States National Institutes of Health; Validation; Veterans; Work; angiogenesis; anti-tumor immune response; cancer type; chemotherapy; combinatorial; efficacy evaluation; gamma secretase; genetic manipulation; high risk; human disease; improved; in vivo; inhibitor; insight; melanoma; melanomagenesis; mouse model; nano-string; novel; novel drug combination; overexpression; patient derived xenograft model; preclinical study; repository; research clinical testing; response; small molecule; success; survivin; targeted treatment; therapeutic target; therapy resistant; transcriptome sequencing; treatment response; tumor; tumor progression; ultraviolet

The overall objective of this study is to determine the therapeutic efficacy of concomitant inhibition of PLK1 and NOTCH against melanoma progression and drug resistance as well as to identify novel signaling mechanisms associated with drug response using two human-relevant melanoma mouse models. The available therapeutic strategies against melanoma have either failed to achieve >25% response in patients, or the responses are short-lived with developing resistance to therapy. For example, BRAF inhibitors Vemurafenib and Dabrafenib were found to achieve significant improvement over chemotherapy and were FDA-approved for melanomas with BRAF-mutations. Even with a combination of Dabrafenib with MEK inhibitor Trametinib (also FDA approved), the patients develop acquired resistance. More recently advancements in immunotherapy have improved melanoma treatment outcomes. Despite the success of immune checkpoint inhibitors durable responses are not seen in all patients due to drug resistance. Therefore, novel mechanism- based combinatorial approaches are needed for an effective management of this neoplasm. Polo-like kinase 1 (PLK1) is a serine/threonine protein kinase that plays a key role in cell proliferation. We have previously reported that PLK1 is significantly overexpressed in melanoma and can be therapeutically targeted. Further, the NOTCH pathway, an evolutionally conserved pathway, which plays important roles in cell fate determination, proliferation, differentiation and survival, has been shown to regulate many aspects of melanomagenesis. NOTCH1 is considered a primary oncogenic factor in melanoma and activation of NOTCH1 and its target genes is linked with metastatic melanoma. Moreover, inhibition of PLK1 or NOTCH has been shown to modulate markers of epithelial mesenchymal transition (EMT) and metastasis. Interestingly, both PLK1 and NOTCH are also linked with drug resistance. Our recently published data suggest that PLK1 and NOTCH expressions have significant positive correlation in melanoma clinical tissues and simultaneous small molecule inhibition of PLK1 and NOTCH by BI 6727 (specific inhibitors of PLK1) and MK-0752 (γ-secretase inhibitor), respectively, caused a significant anti-proliferative response in multiple melanoma cell lines, warranting further pre-clinical testing in in vivo melanoma models. This data together with other published studies provide a strong scientific premise for our proposed hypothesis that combined inhibition of PLK1 and NOTCH will be therapeutically superior for the management of melanoma. We will challenge this hypothesis in two specific aims. In Aim 1, we will determine the in vivo therapeutic efficacy and mechanism of concomitant inhibition of PLK1 (by BI 6727 or PCM-075) and NOTCH (by MK-0752) on melanoma progression and metastasis in genetically engineered Braf-Pten melanoma mouse model, which recapitulates human disease progression from localized to metastatic disease. In Aim 2, we will determine therapeutic efficacy and mechanism of concomitant inhibition of PLK1 and NOTCH against melanoma drug resistant using patient derived xenograft (PDX) model, which conserve original tumor characteristics and offer relevant predictive insights into clinical outcomes, for direct relevance to clinical management of melanoma. Additionally, we will determine the novel mechanisms associated with treatment response using mutiple techniques such as Nanostring PanCancer Progression Panel, global proteomics and RNA-seq analyses in tumor samples followed by validation using RT-qPCR and ProteinSimple analyses. Overall, our study is expected to provide mechanistic insights and rationale for clinical testing of the combined PLK1-NOTCH inhibition to obtain superior anti-melanoma response and overcome resistance. Our proposed work is relevant and significant to the Veterans because melanoma is the fifth most diagnosed malignancy among Veterans, and the fact that the US military has been engaged, in missions all over the world, many US military personnel, who are deployed to high ultraviolet (UV) climates in tropical and subtropical zones are potentially at a higher risk for melanoma.

这项研究的总体目的是确定伴随抑制PLK1的治疗效率 和针对黑色素瘤进展和耐药性的缺陷以及识别新型信号 使用两个与人类相关的黑色素瘤小鼠模型相关的机制。 针对黑色素瘤的可用治疗策略要么未能达到> 25％的反应 患者或反应是短暂的，具有对治疗的耐药性。例如，BRAF抑制剂 发现维美富尼和达布拉尼在化学疗法方面取得了显着改善， FDA批准用于黑色素瘤，并具有BRAF突变。即使将Dabrafenib与MEK抑制剂结合在一起 Trametinib（也获得了FDA批准），患者会产生获得性耐药性。最近的进步 免疫疗法改善了黑色素瘤治疗结果。尽管Immunocheckpoint取得了成功 由于耐药性，所有患者均未看到抑制剂耐用反应。因此，新型机制 - 需要基于组合方法来有效地管理这种肿瘤。 polo样激酶1 （PLK1）是一种丝氨酸/苏氨酸蛋白激酶，在细胞增殖中起关键作用。我们以前有 据报道，PLK1在黑色素瘤中明显过表达，并且可以被热靶向。此外， Notch途径是一种进化配置的途径，在细胞命运中起着重要作用 确定，增殖，分化和生存已被证明可以调节 黑素作仿。 Notch1被认为是黑色素瘤和Notch1激活中的主要致癌因子 它的靶基因与转移性黑色素瘤有关。此外，抑制PLK1或Notch已经 显示以调节上皮间质转变（EMT）和转移的标记。有趣的是，两者俩 PLK1和Notch也与耐药性有关。我们最近发布的数据表明PLK1和 Notch表达在黑色素瘤临床组织中具有显着的正相关性，而简单的小 BI 6727（PLK1的特定抑制剂）和MK-0752（γ-分泌酶）对PLK1和NATCH的分子抑制 抑制剂）分别在多个黑色素瘤细胞系中引起明显的抗增殖反应， 警告体内黑色素瘤模型中进一步的临床前测试。这些数据以及其他已发布的 研究为我们提出的假设提供了强大的科学前提，该假设将PLK1和 Notch将在黑色素瘤的治疗中具有热优势。我们将在 两个具体的目标。在AIM 1中，我们将确定体内治疗效率和伴随的机制 抑制PLK1（通过BI 6727或PCM-075）和Notch（通过MK-0752）对黑色素瘤进展和 一般设计的BRAF-PTEN黑色素瘤小鼠模型的转移，该模型概括了人类疾病 从局部疾病到转移性疾病的发展。在AIM 2中，我们将确定治疗效率和 使用患者对PLK1和NOTCH对黑色素瘤耐药性抗黑色素瘤的抑制机制 衍生的异种移植（PDX）模型，该模型保留原始肿瘤特征并提供相关的预测 对临床结果的见解，与黑色素瘤的临床管理直接相关。此外，我们会的 确定使用Mutiple技术（例如 肿瘤样品中的纳米弦pancancer进度面板，全局蛋白质组学和RNA-Seq分析 然后使用RT-QPCR和ProteInsimple分析进行验证。总体而言，我们的研究有望提供 机械洞察力和基本原理，用于联合PLK1-NOTCH抑制的临床测试以获得 上抗黑色素瘤反应并克服抗性。我们提出的工作与 退伍军人是因为黑色素瘤是退伍军人中最受诊断出的恶性肿瘤，这一事实是 美国军方已在世界各地的许多美国军事人员中订婚，他们被部署到 热带和亚热带地区的高紫外线（紫外线）攀岩可能面临黑色素瘤的风险更高。