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.

本研究的总体目的是确定PLK 1的伴随抑制的治疗效果 和NOTCH针对黑色素瘤进展和耐药性以及识别新型信号传导 使用两种人类相关的黑色素瘤小鼠模型研究与药物反应相关的机制。 针对黑色素瘤的现有治疗策略要么未能实现>25%的缓解， 患者，或反应是短暂的，对治疗产生耐药性。例如，BRAF抑制剂 发现威罗菲尼和达拉非尼相对于化疗实现了显著改善， FDA批准用于BRAF突变的黑色素瘤即使使用达拉非尼与MEK抑制剂的组合， 曲美替尼（也是FDA批准的），患者会产生获得性耐药。最近的进展， 免疫疗法改善了黑色素瘤治疗结果。尽管免疫检查点的成功 由于耐药性，在所有患者中未观察到抑制剂持久反应。因此，新机制- 需要基于组合的方法来有效管理这种肿瘤。polo样激酶1 PLK 1是一种丝氨酸/苏氨酸蛋白激酶，在细胞增殖中起关键作用。我们先前已经 报道了PLK 1在黑素瘤中显著过表达，并且可以是治疗靶向的。此外，本发明还 NOTCH通路是一种进化上保守的通路，在细胞命运中起重要作用 决定，增殖，分化和存活，已被证明可以调节许多方面， 黑色素瘤形成NOTCH 1被认为是黑色素瘤中的主要致癌因子， 其靶基因与转移性黑色素瘤有关。此外，PLK 1或NOTCH的抑制已经被证实是有效的。 显示调节上皮间质转化（EMT）和转移的标志物。有趣的是，两者 PLK 1和NOTCH也与耐药性有关。我们最近发表的数据表明，PLK 1和 NOTCH在黑色素瘤临床组织中的表达呈显著正相关， BI 6727（PLK 1特异性抑制剂）和MK-0752（γ-分泌酶）对PLK 1和NOTCH的分子抑制 抑制剂）分别在多种黑素瘤细胞系中引起显著的抗增殖反应， 在体内黑色素瘤模型中进行进一步的临床前试验。这些数据与其他出版的 研究为我们提出的假设提供了强有力的科学前提，即PLK 1和 对于黑色素瘤的管理，NOTCH将在治疗上具有上级优势。我们将挑战这一假设， 两个具体目标。在目标1中，我们将确定伴随的药物的体内治疗功效和机制。 PLK 1（BI 6727或PCM-075）和NOTCH（MK-0752）对黑色素瘤进展的抑制作用， 在基因工程Braf-Pten黑色素瘤小鼠模型中的转移，其重现了人类疾病 从局限性疾病进展为转移性疾病。在目标2中，我们将确定治疗效果， PLK 1和NOTCH对黑色素瘤耐药性的共同抑制机制 衍生的异种移植物（PDX）模型，其保留了原始肿瘤特征并提供相关的预测 深入了解临床结果，与黑素瘤的临床管理直接相关。此外，我们将 使用多种技术确定与治疗反应相关的新机制，例如 Nanostring PanCancer Progression Panel，肿瘤样本中的全球蛋白质组学和RNA-seq分析 随后使用RT-qPCR和ProteinSimple分析进行验证。总的来说，我们的研究预计将提供 联合PLK 1-NOTCH抑制的临床试验的机制见解和原理，以获得 上级抗黑色素瘤反应和克服耐药性。我们建议的工作对以下方面具有相关性和重要性： 退伍军人，因为黑色素瘤是退伍军人中第五大诊断恶性肿瘤，事实上， 美国军方在世界各地执行任务，许多美国军事人员被部署到 热带和亚热带地区的高紫外线（UV）气候可能具有更高的黑色素瘤风险。