Molecular approaches to overcome intrinsic drug resistance in BRAF and NRAS-mutan

克服 BRAF 和 NRAS-mutan 固有耐药性的分子方法

• 批准号: 8826711
• 负责人: Jessie Villanueva
• 金额: $ 15.08万
• 依托单位: WISTAR INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-04-01 至 2016-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8826711
• 关键词: Affect; Aftercare; Apoptosis; Area; Attenuated; Automobile Driving; Autophagocytosis; BRAF gene; Biochemical; Biological; Bypass; Cell Cycle; Cell Line; Cell Survival; Cells; Clinical; Clinical Trials; Computational Biology; Data; Dependency; Development; Diagnosis; Disease; Drug Combinations; Drug resistance; EGFR gene; Family member; Feedback; Fostering; Funding; Future; Goals; Health; IGF1 gene; Immunocompromised Host; In Vitro; Incidence; Innovative Therapy; Knowledge; Laboratories; Life; MAP Kinase Gene; MEK inhibition; MEKs; Malignant Neoplasms; Mediating; Mediator of activation protein; Medical; Melanoma Cell; Mentors; Metastatic Melanoma; Mitogen-Activated Protein Kinase Inhibitor; Molecular; Molecular Biology; Mus; Mutate; Mutation; NRAS gene; Neoplasm Metastasis; Neoplasms; Oncogenes; Oncogenic; Outcome; PDGFRB gene; Pathway interactions; Patients; Proteomics; Proto-Oncogene Proteins c-akt; Public Health; Receptor Protein-Tyrosine Kinases; Recurrent disease; Relative (related person); Reporting; Research; Research Personnel; Resistance; Resistance development; Role; SEER Program; Sampling; Scientist; Signal Pathway; Signal Transduction; Survival Rate; System; Systems Analysis; Testing; The Wistar Institute; Therapeutic; Therapeutic Effect; Training; Tumor-Derived; Up-Regulation; Work; Xenograft Model; Xenograft procedure; base; combinatorial; design; effective therapy; experience; improved; inhibitor/antagonist; melanoma; mutant; overexpression; prevent; professor; programs; response; success; targeted treatment; therapeutic target; therapy resistant; treatment strategy; tumor; tumor xenograft; tumorigenesis

DESCRIPTION (provided by applicant): The applicant is an assistant professor at Molecular and Cellular Oncogenesis Program at the Wistar Institute. Her overall goal is to develop effective and long-lasting treatment strategies for melanoma. This proposal serves the applicant's long-term objective of improving outcomes for melanoma patients by investigating the mechanisms of intrinsic resistance to targeted therapy in BRAF-V600E and NRAS mutant melanomas. The candidate will pursue training in research areas that are essential to foster a research niche and become an independent academic cancer researcher. A team of senior scientific advisors has been assembled to formally mentor the applicant by providing training and expertise in new research areas including proteomics, systems and computational biology, and autophagy. The mentoring team is committed to support the development of the applicant into a fully funded independent scientist. Melanoma is a highly aggressive disease with limited therapeutic options. Despite the success of BRAF inhibitors for the treatment of melanoma, responses are transient and most patients develop resistance. Therefore, a major challenge is to understand, overcome, and prevent drug resistance. This application proposes to investigate the mechanisms of intrinsic drug resistance, an important unmet medical need affecting a large subset of melanoma patients, for whom no effective therapies are available. The proposal focuses on NRAS mutant melanomas and BRAF-V6000E mutant melanomas that are intrinsically resistant to BRAF inhibitors. The overall hypothesis is that uncovering the mechanisms underlying intrinsic drug resistance in melanoma will allow designing more effective and longer lasting therapies. The applicant postulates that inhibition of BRAF induces rewiring of the system's signaling networks leading to receptor tyrosine kinase (RTK) activation and attenuated dependency on BRAF. To test this hypothesis, the expression and activation of RTKs will be evaluated in melanoma cell lines and patient-derived tumor samples grown in immunocompromised mice (patient-derived xenografts; PDX) following BRAF inhibition. Mechanistic studies will be performed to determine how inhibition of MAPK leads to RTK activation. Based on these studies rational drug combination strategies will be evaluated in melanoma cells grown as 3D tumor spheroids and in PDX models. The second working hypothesis is that by identifying critical downstream NRAS effectors, melanomas harboring mutations in NRAS can be targeted. The objective is to define the biochemical and biological consequences of silencing NRAS in melanoma cells that harbor mutations in this oncogene. To achieve this goal a candidate and an unbiased proteomic based approaches will be undertaken to identify NRAS effectors that are essential for survival of NRAS mutant melanomas. Collectively, this knowledge will be critical to develop new strategies that eliminate nearly all melanoma cells and inform the design of future clinical trials.

描述（由申请人提供）：申请人是 Wistar 研究所分子和细胞肿瘤发生项目的助理教授。她的总体目标是开发有效且持久的黑色素瘤治疗策略。该提案通过研究 BRAF-V600E 和 NRAS 突变黑色素瘤对靶向治疗的内在耐药机制，服务于申请人改善黑色素瘤患者预后的长期目标。候选人将接受研究领域的培训，这对于培育研究利基并成为独立的学术癌症研究员至关重要。一支由高级科学顾问组成的团队已组建完毕，通过提供蛋白质组学、系统和计算生物学以及自噬等新研究领域的培训和专业知识来正式指导申请人。指导团队致力于支持申请人发展成为一名获得全额资助的独立科学家。 黑色素瘤是一种高度侵袭性的疾病，治疗选择有限。尽管 BRAF 抑制剂在治疗黑色素瘤方面取得了成功，但反应是短暂的，并且大多数患者会产生耐药性。因此，一个重大挑战是了解、克服和预防耐药性。该申请旨在研究内在耐药性的机制，这是一种重要的未满足的医疗需求，影响着一大批黑色素瘤患者，而这些患者目前还没有有效的治疗方法。该提案重点关注对 BRAF 抑制剂具有内在耐药性的 NRAS 突变黑色素瘤和 BRAF-V6000E 突变黑色素瘤。总体假设是，揭示黑色素瘤内在耐药性的机制将有助于设计更有效、更持久的疗法。申请人假设BRAF的抑制诱导系统信号网络的重新布线，导致受体酪氨酸激酶(RTK)激活并减弱对BRAF的依赖性。为了检验这一假设，我们将在 BRAF 抑制后，在免疫功能低下的小鼠（患者来源的异种移植物；PDX）中生长的黑色素瘤细胞系和患者来源的肿瘤样本中评估 RTK 的表达和激活。将进行机制研究以确定 MAPK 的抑制如何导致 RTK 激活。基于这些研究，将在生长为 3D 肿瘤球体的黑色素瘤细胞和 PDX 模型中评估合理的药物组合策略。第二个工作假设是，通过识别关键的下游 NRAS 效应子，可以靶向携带 NRAS 突变的黑色素瘤。目的是确定在含有该癌基因突变的黑色素瘤细胞中沉默 NRAS 的生化和生物学后果。为了实现这一目标，将采用候选方法和基于公正的蛋白质组学的方法来识别对 NRAS 突变黑色素瘤的生存至关重要的 NRAS 效应子。总的来说，这些知识对于制定消除几乎所有黑色素瘤细胞的新策略至关重要，并为未来临床试验的设计提供信息。