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.

本研究的总体目标是确定同时抑制PLK1的治疗效果。 和Notch抗黑色素瘤进展和耐药性以及识别新的信号 使用两个与人类相关的黑色素瘤小鼠模型与药物反应相关的机制。 现有的治疗黑色素瘤的策略要么未能在#年达到25%的反应 患者，或者反应是短暂的，并对治疗产生抵抗力。例如，BRAF抑制剂 维莫拉非尼和达普拉非尼被发现比化疗有显著改善，并 FDA批准用于BRAF突变的黑色素瘤。即使联合使用达普拉非尼和MEK抑制剂 曲美替尼(也是FDA批准的)，患者会产生获得性耐药性。最近，在 免疫疗法改善了黑色素瘤的治疗结果。尽管免疫检查站取得了成功 由于耐药性，并不是所有的患者都能看到抑制剂的持久反应。因此，新的机制-- 这种肿瘤的有效治疗需要基于联合治疗的方法。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。我们之前已经 报道称，PLK1在黑色素瘤中显著过度表达，可以作为治疗的靶点。此外， Notch途径是一条进化上保守的途径，在细胞命运中起着重要作用 决心、增殖、分化和存活，已经被证明在许多方面调节 黑色素瘤形成。NOTCH1被认为是黑色素瘤的主要致癌因子和NOTCH1的激活 它的靶基因与转移性黑色素瘤有关。此外，对PLK1或缺口的抑制已经被 被认为是调节上皮间充质转化(EMT)和转移的标志物。有趣的是，两者 PLK1和Noch也与耐药有关。我们最近公布的数据表明，PLK1和PLK1 Notch在黑色素瘤临床组织和小黑色素瘤中的表达呈显著正相关 BI6727(PLK1特异性抑制剂)和MK-0752(γ分泌酶)对PLK1和Noch的分子抑制作用 抑制剂)分别在多个黑色素瘤细胞系中引起显著的抗增殖反应， 保证在体内黑色素瘤模型中进行进一步的临床前测试。这些数据与其他已发布的数据一起 研究为我们提出的假设提供了强有力的科学前提，即PLK1和PLK1的抑制 对于黑色素瘤的治疗而言，NOCK将具有更好的治疗效果。我们将在以下方面挑战这一假设 有两个明确的目标。在目标1中，我们将确定伴随剂的体内治疗效果和机制。 PLK1(BI 6727或PCM-075)和Noch(MK-0752)抑制黑色素瘤进展和 重述人类疾病的基因工程BRAF-Pten黑色素瘤小鼠模型的转移 从局限性疾病进展到转移性疾病。在目标2中，我们将确定治疗效果和 PLK1和Noch联合抑制黑色素瘤耐药机制的研究 衍生异种移植(PDX)模型，它保留了原始肿瘤的特征并提供了相关的预测 对临床结果的洞察，与黑色素瘤的临床治疗直接相关。此外，我们还将 使用多种技术确定与治疗反应相关的新机制，例如 纳米串泛癌进展小组、全球蛋白质组学和肿瘤样本中的RNA-seq分析 然后用RT-qPCR和ProteinSimple分析进行验证。总体而言，我们的研究有望提供 PLK1-缺口抑制联合临床试验的机理见解和理论基础 卓越的抗黑色素瘤反应，克服耐药性。我们提出的工作对以下方面具有重要意义 退伍军人是因为黑色素瘤是退伍军人中第五大被诊断为恶性肿瘤的疾病，而事实上 美国军方在世界各地执行任务，部署了许多美军人员， 热带和亚热带地区的高紫外线(UV)气候可能有更高的黑色素瘤风险。