Targeting Abl kinases in BRAF-driven melanomas

靶向 BRAF 驱动的黑色素瘤中的 Abl 激酶

• 批准号: 10221629
• 负责人: RINA PLATTNER
• 金额: $ 34.85万
• 依托单位: UNIVERSITY OF KENTUCKY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-08-13 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10221629
• 关键词: Automobile Driving; BRAF gene; Binding; Binding Sites; Biochemical; Biological; Cell Cycle Progression; Cell Line; Cells; Chronic; Clinical Research; Complex; Data; Development; Disease; Disease Resistance; Disease remission; Drug Targeting; Environment; Experimental Designs; FDA approved; Genetic Engineering; Goals; Growth; Immunohistochemistry; Immunotherapy; Incidence; Intrinsic drive; Knock-in; Lead; Link; Malignant Neoplasms; Masks; Mass Spectrum Analysis; Mediating; Melanoma Cell; Messenger RNA; Metastatic Melanoma; Molecular; Mucous Membrane; Mutation; Nature; Neoplasm Metastasis; Nuclear; Nuclear Localization Signal; Oncologist; Outcome; PTEN gene; Pathway interactions; Patients; Pharmacology; Phenotype; Phosphorylation; Phosphotransferases; Physiological; Play; Positioning Attribute; Proteins; Proto-Oncogene Protein c-kit; RNA Interference; Refractory Disease; Repression; Research; Resistance; Resistance development; Role; Signal Transduction; Solid Neoplasm; Sun Exposure; TWIST1 gene; Testing; Therapeutic; Threonine; Time; Tissue Microarray; Work; Xenograft Model; Xenograft procedure; base; design; efficacy testing; experimental study; gain of function; genetic approach; in vivo; in vivo Model; inhibitor/antagonist; innovation; insight; leukemia; loss of function; melanocyte; melanoma; mouse model; mutant; novel; novel drug combination; novel marker; prevent; recruit; src-Family Kinases; success; targeted agent; therapy resistant; three dimensional cell culture; transcription factor; treatment response; tumor growth; young woman

Despite the success of new targeted agents and immunotherapies, metastatic melanoma remains an incurable disease for many patients. Although BRAF inhibitors (BRAFi) show promise for reducing metastatic burden, some melanomas are intrinsically resistant, and most responding patients eventually acquire resistance. These data underline the importance of identifying new markers for treatment response and novel drug combinations for treating metastatic and BRAFi-resistant disease. We show that Abl/Arg non-receptor tyrosine kinases are highly expressed in melanoma, and are activated in a subset (40-60%). Moreover, SFKs and BRAFV600E, the most common genetic alteration in melanoma, both contribute to Abl/Arg activation in melanoma cells. Once activated, Abl/Arg promote proliferation, survival, a switch in EMT transcription factor expression, invasion, and metastasis and bidirectionally regulate and potentiate BRAFV600E signaling. Moreover, inhibitors of Abl/Arg and a parallel pathway, which drives intrinsic BRAFi resistance (PI3K/Akt/PTEN), cooperate to prevent melanoma viability and tumor growth. Based on these findings, we hypothesize that BRAFV600E/ERK and SFKs cooperatively activate Abl/Arg, which synergize with Akt to drive melanoma development, progression, phenotypic switching and subsequent resistance. We propose a comprehensive hypothesis-driven experimental design that will establish Abl/Arg as novel and exploitable drug targets. Aim 1 will define the mechanism by which Abl/Arg are activated in melanoma. To achieve our objective, biochemical, molecular biological and cell biological approaches using melanocytes, melanoma cell lines, tissue microarrays, pharmacological inhibitors/RNAi, mass spectrometry, and immunohistochemistry will be used to test the prediction that BRAFV600E/ERK-mediated phosphorylation prevents nuclear targeting of Abl and facilitates activation of Abl/Arg by SFKs. In Aim 2, 2D/3D culture, rescue experiments and xenograft approaches will be used to identify the mechanism by which Abl/Arg drive the EMT transcription factor switch, potentiate BRAFV600E signaling, and promote acquired BRAFi resistance. Finally, in Aim 3, genetically engineered (GEM) and xenograft mouse models as well as loss- and gain-of-function approaches will be used to test the prediction that Abl/Arg cooperate with activated Akt, in mutant PTEN melanomas, to promote melanoma growth/metastasis and BRAFi/MEKi resistance. Data obtained from this project not only will allow us to gain important insight into fundamental mechanisms by which Abl/Arg are activated in melanoma, which likely is applicable to other solid tumors, but also may lead to clinical studies testing the efficacy of Akt inhibitors in combination with Abl/Arg inhibitors for treating mutant BRAF/PTEN melanomas, which often are resistant to current therapeutic approaches.

尽管新的靶向药物和免疫疗法取得了成功，但转移性黑色素瘤仍然是一种无法治愈的疾病。 对于很多患者来说，虽然BRAF抑制剂（BRAFi）显示出减少转移负荷的前景， 一些黑色素瘤具有内在的抗性，并且大多数有反应的患者最终获得抗性。这些 数据强调了识别治疗反应的新标志物和新型药物组合的重要性 用于治疗转移性和BRAFi抗性疾病。我们发现Abl/Arg非受体酪氨酸激酶是 在黑色素瘤中高度表达，并且在一个亚组中被激活（40-60%）。此外，SFK和BRAFV 600 E， 黑色素瘤中最常见的遗传改变，都有助于黑色素瘤细胞中Abl/Arg的激活。一旦 激活，Abl/Arg促进增殖，存活，EMT转录因子表达的开关，侵袭， 转移和双向调节和增强BRAFV 600 E信号传导。此外，Abl/Arg和 驱动内在BRAFi抗性（PI 3 K/Akt/PTEN）的平行通路合作预防黑色素瘤 存活力和肿瘤生长。基于这些发现，我们推测BRAFV 600 E/ERK和SFKs可能与细胞凋亡有关。 协同激活Abl/Arg，Abl/Arg与Akt协同作用以驱动黑色素瘤的发展、进展， 表型转换和随后的抗性。我们提出了一个全面的假设驱动的 实验设计，将建立Abl/Arg作为新的和可开发的药物靶标。目标1将定义 黑色素瘤中Abl/Arg被激活的机制。为了达到我们的目标，生化，分子 使用黑素细胞，黑色素瘤细胞系，组织微阵列， 药理学抑制剂/RNAi、质谱和免疫组织化学将用于检测 预测BRAFV 600 E/ERK介导的磷酸化阻止Abl的核靶向并促进 SFKs激活Abl/Arg。在目标2中，2D/3D培养、挽救实验和异种移植方法将被 用于鉴定Abl/Arg驱动EMT转录因子开关的机制， BRAFV 600 E信号传导，并促进获得性BRAFi抗性。最后，在目标3中，基因工程（GEM） 和异种移植小鼠模型以及功能丧失和获得方法将用于测试 在突变型PTEN黑色素瘤中Abl/Arg与激活的Akt协同促进黑色素瘤的预测 生长/转移和BRAFi/MEKi抗性。从这个项目中获得的数据不仅可以让我们获得 对Abl/Arg在黑色素瘤中被激活的基本机制的重要见解， 这不仅适用于其他实体瘤，而且还可能导致临床研究测试Akt抑制剂在 与Abl/Arg抑制剂组合用于治疗突变型BRAF/PTEN黑色素瘤，所述突变型BRAF/PTEN黑色素瘤通常对 目前的治疗方法。