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: 9762875
• 负责人: RINA PLATTNER
• 金额: $ 33.8万
• 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/9762875
• 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.

尽管新靶向剂和免疫疗法成功，但转移性黑色素瘤仍然是无法治愈的 许多患者的疾病。尽管BRAF抑制剂（BRAFI）显示出减轻转移负担的希望，但 一些黑色素瘤本质上具有抗性，大多数反应的患者最终都会获得抗药性。这些 数据强调了识别新标记以进行治疗反应和新型药物组合的重要性 用于治疗转移和抗Brafi疾病。我们表明ABL/ARG非受体酪氨酸激酶是 在黑色素瘤中高表达，并在子集中激活（40-60％）。此外，SFK和BRAFV600E， 黑色素瘤中最常见的遗传改变，都导致黑色素瘤细胞中的ABL/ARG激活。一次 激活的ABL/ARG促进增殖，生存，EMT转录因子表达中的转换，入侵和 转移和双向调节和增强BRAFV600E信号传导。此外，ABL/ARG的抑制剂和 平行途径，它驱动固有的BRAFI耐药性（PI3K/AKT/PTEN），以防止黑色素瘤 活力和肿瘤生长。基于这些发现，我们假设BRAFV600E/ERK和SFK 合作激活ABL/ARG，该AB/ARG与AKT协同以推动黑色素瘤的发展，进展， 表型切换和随后的电阻。我们提出了一个全面的假设驱动的 实验设计将建立ABL/ARG作为新颖和可剥削的药物靶标。 AIM 1将定义 在黑色素瘤中激活ABL/ARG的机制。为了实现我们的目标，生化，分子 使用黑色素细胞，黑色素瘤细胞系，组织微阵列的生物学和细胞生物学方法， 药理抑制剂/RNAI，质谱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的核靶向和促进 SFK激活ABL/ARG。在AIM 2，2D/3D文化中，救援实验和异种移植方法将是 用于识别ABL/ARG驱动EMT转录因子开关的机制，增强 BRAFV600E信号传导，并促进获得的BRAFI电阻。最后，在AIM 3中，基因工程（宝石） 异种移植小鼠模型以及功能障碍方法将用于测试 预测ABL/ARG与激活的Akt合作，在突变体Pten黑色素瘤中促进黑色素瘤 生长/转移和BRAFI/MEKI抗性。从该项目获得的数据不仅允许我们获得 对ABL/ARG在黑色素瘤中激活ABL/ARG的基本机制的重要见解，这可能是 适用于其他实体瘤，但也可能导致临床研究测试Akt抑制剂在 与ABL/ARG抑制剂结合使用用于治疗突变体BRAF/PTEN黑色素瘤的抑制剂，通常对 当前的治疗方法。