Adaptive signaling exposes a therapeutic vulnerability in NRAS melanomas

适应性信号传导暴露了 NRAS 黑色素瘤的治疗脆弱性

• 批准号: 8621637
• 负责人: MICHAEL J. WEBER
• 金额: $ 20.62万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-01 至 2016-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8621637
• 关键词: 1-Phosphatidylinositol 3-Kinase; Advanced Malignant Neoplasm; Apoptosis; Autophagocytosis; BRAF gene; Back; Calcium; Cell Cycle Arrest; Cell Line; Clinical; Clinical Trials; Combined Modality Therapy; Complement; Cutaneous Melanoma; Cyclic AMP-Dependent Protein Kinases; Data; Defense Mechanisms; Dependency; Development; Disease; Drug Combinations; Drug Targeting; Effectiveness; FDA approved; Family; Family member; Foundations; Gene Expression; Gene Expression Profile; Genetic Epistasis; Genomics; Growth; Knowledge; Lead; Link; MAPK7 gene; Malignant Neoplasms; Melanoma Cell; Methodology; Mitochondria; Mitogen-Activated Protein Kinases; Molecular; Mutate; Oncogenes; Oxygen; Pathway Analysis; Pathway interactions; Patients; Pharmaceutical Preparations; Phase; Phenocopy; Phosphotransferases; Proteins; Proteomics; RAF1 gene; RNA Interference; Recurrence; Research; Resistance; Sampling; Signal Pathway; Signal Transduction; Techniques; Testing; Therapeutic; Therapeutic Agents; Toxic effect; Universities; Virginia; Work; analog; base; cancer cell; clinical application; computerized tools; cytotoxic; cytotoxicity; high throughput screening; in vivo; inhibitor/antagonist; insight; killings; lapatinib; melanoma; mutant; public health relevance; receptor; research study; resistance mechanism; response; small molecule; therapeutic target; tool

DESCRIPTION (provided by applicant): Analysis of adaptive signaling in response to targeted therapies can provide insight into mechanisms of resistance and appropriate combinations of therapeutic agents. In a screen for adaptive resistance mechanisms in BRAF mutant melanomas, we made a serendipitous discovery that may provide a path to targeted therapy for melanomas driven by mutant NRAS, a disease for which there are few options. We found that a combination of a RAF inhibitor (PLX4720) and a HER-family inhibitor (lapatinib) could kill NRAS mutant melanomas that were resistant to either drug alone. The mechanism of killing did not require inhibition of the MAP Kinase or the PI3 Kinase pathways. The data strongly suggest that either RAF or a HER family receptor (or both) are engaged in non-canonical signaling activity that provides a previously undetected cytotoxic vulnerability. This proposal aims to identify the molecular basis for that non-canonical signaling, and determine the most appropriate setting for in vivo studies and a clinical trial. Aim 1. We will analyze the signaling network that links BRAF, NRAS and HER family members to identify signaling nodes critical for the PLX4720-lapatinib synergy. We will utilize proteomic techniques (Reverse Phase Protein Micro Arrays; RPPA) and gene expression arrays to determine the adaptive responses to drug treatments made in the phosphoproteome and the transcriptome. We also will analyze the mode(s) of cytotoxicity induced by combined therapy (apoptosis, cell cycle arrest, autophagy). These responses will be analyzed using publicly available computational tools as well as integrated pathway analysis tools developed by our collaborators at the University of Virginia. We will explicitly examine our -omic analyses for hypothesized mechanisms including functional interactions of CRAF or BRAF with mitochondria, Protein Kinase A and the spindle assembly checkpoint; and of HER family kinases with the HIPPO pathway, RAC, PAK, Reactive Oxygen, Calcium, and ERK5. Aim 2. We will test the functional significance of signaling nodes identified in Aim 1. We will perform epistasis experiments, complementing cytotoxicity with RNAi and rescuing inhibited growth with ORFeome expression. We also will determine whether appropriate small molecules can phenocopy the cytotoxic effects of the PLX4720-lapatinib combination. Adaptive homeostatic responses are a major confounder for targeted therapies, and the information obtained here will provide a strong foundation for therapeutic application with FDA-approved drugs and validated targets.

描述(由申请人提供)：对靶向治疗反应的适应性信号的分析可以提供对耐药机制和治疗药物的适当组合的洞察。在对BRAF突变黑色素瘤适应性耐药机制的筛选中，我们有了一个偶然的发现，可能为突变NRAS驱动的黑色素瘤提供靶向治疗的途径，这种疾病几乎没有选择。我们发现，RAF抑制剂(PLX4720)和HER家族抑制剂(Lapatinib)的组合可以杀死单独对这两种药物耐药的NRAS突变黑色素瘤。这种杀伤机制不需要抑制MAP激酶或PI3激酶通路。这些数据强烈地表明，RAF或HER家族受体(或两者)参与了非规范的信号活动，提供了以前未检测到的细胞毒性脆弱性。这项建议旨在确定非规范信号的分子基础，并确定体内研究和临床试验的最合适环境。目的1.我们将分析连接BRAF、NRAS和她的家庭成员的信号网络，以确定PLX4720-Lapatinib协同作用的关键信号节点。我们将利用蛋白质组学技术(反相蛋白质微阵列；RPPA)和基因表达阵列来确定在磷蛋白质组和转录组中进行的药物治疗的适应性反应。我们还将分析联合治疗(细胞凋亡、细胞周期停滞、自噬)所引起的细胞毒性的模式(S)。这些响应将使用公开可用的计算工具以及由我们在弗吉尼亚大学的合作者开发的集成路径分析工具进行分析。我们将明确审查对假设机制的经济学分析，包括CRAF或BRAF与线粒体、蛋白激酶A和纺锤体组装检查点的功能相互作用；以及她的家族激酶与河马途径、RAC、PAK、活性氧、钙和ERK5的功能相互作用。目的2.我们将测试目标1中确定的信号节点的功能意义。我们将进行上位性实验，用RNAi补充细胞毒性，并用ORFeome表达拯救受抑制的生长。我们还将确定适当的小分子是否可以复制PLX4720-拉帕替尼联合的细胞毒性效应。适应性内环境平衡反应是靶向治疗的一个主要混淆因素，这里获得的信息将为FDA批准的药物和有效靶点的治疗应用提供坚实的基础。