Targeting Abl kinases in BRAF-driven melanomas

靶向 BRAF 驱动的黑色素瘤中的 Abl 激酶

基本信息

批准号：
10449267
负责人：
RINA PLATTNER
金额：
$ 34.15万
依托单位：
UNIVERSITY OF KENTUCKY
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-08-13 至 2024-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10449267
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 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

项目摘要

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许多患者的疾病。尽管 BRAF 抑制剂（BRAFi）有望减少转移负担，一些黑色素瘤具有内在耐药性，大多数有反应的患者最终会获得耐药性。这些数据强调了识别治疗反应新标志物和新药物组合的重要性用于治疗转移性和 BRAFi 耐药性疾病。我们证明 Abl/Arg 非受体酪氨酸激酶是在黑色素瘤中高度表达，并在一个亚群（40-60%）中被激活。此外，SFK 和 BRAFV600E 黑色素瘤中最常见的基因改变都有助于黑色素瘤细胞中 Abl/Arg 的激活。一次激活后，Abl/Arg 可促进增殖、存活、EMT 转录因子表达、侵袭和转移的转换转移并双向调节和增强 BRAFV600E 信号传导。此外，Abl/Arg 抑制剂和驱动内在 BRAFi 抵抗 (PI3K/Akt/PTEN) 的并行途径，共同预防黑色素瘤活力和肿瘤生长。基于这些发现，我们假设 BRAFV600E/ERK 和 SFK 协同激活 Abl/Arg，与 Akt 协同作用，驱动黑色素瘤的发生、进展，表型转换和随后的耐药性。我们提出了一个全面的假设驱动的实验设计将 Abl/Arg 确立为新颖且可开发的药物靶点。目标 1 将定义 Abl/Arg 在黑色素瘤中被激活的机制。为了实现我们的目标，生化、分子使用黑素细胞、黑素瘤细胞系、组织微阵列的生物学和细胞生物学方法，药物抑制剂/RNAi、质谱法和免疫组织化学将用于测试预测 BRAFV600E/ERK 介导的磷酸化可阻止 Abl 的核靶向并促进 SFK 激活 Abl/Arg。在目标 2 中，将进行 2D/3D 培养、拯救实验和异种移植方法用于鉴定 Abl/Arg 驱动 EMT 转录因子开关、增强的机制 BRAFV600E信号传导，促进获得性BRAFi耐药。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预测在突变 PTEN 黑色素瘤中，Abl/Arg 与激活的 Akt 合作，促进黑色素瘤生长/转移和 BRAFi/MEKi 抗性。从这个项目中获得的数据不仅能让我们获得对 Abl/Arg 在黑色素瘤中激活的基本机制的重要见解，这可能是适用于其他实体瘤，而且还可能导致临床研究测试 Akt 抑制剂的功效与 Abl/Arg 抑制剂联合治疗突变型 BRAF/PTEN 黑色素瘤，这种肿瘤通常对目前的治疗方法。