Mechanistic Basis for ERK in driving KRAS-dependent pancreatic cancer

ERK 驱动 KRAS 依赖性胰腺癌的机制基础

• 批准号: 10739653
• 负责人: Jennifer E Klomp
• 金额: $ 11.14万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10739653
• 关键词: Acute; Address; Automobile Driving; CRISPR library; CRISPR screen; CRISPR/Cas technology; Cancer Etiology; Cells; Cessation of life; Clinical; Combined Modality Therapy; Complex; Computer Analysis; Cytoplasm; Data; Development; Ectopic Expression; Engineering; Failure; Genetic; Goals; Growth; Growth Factor; In Vitro; KRAS2 gene; Knowledge; Libraries; Location; MAPK1 gene; MAPK3 gene; MEKs; Malignant Neoplasms; Malignant neoplasm of lung; Malignant neoplasm of pancreas; Mass Spectrum Analysis; Mentors; Mitogen-Activated Protein Kinase Inhibitor; Mitogen-Activated Protein Kinases; Mutate; Mutation; Normal tissue morphology; Nuclear; Nuclear Export; Oncogenes; Outcome; Pancreatic Ductal Adenocarcinoma; Pathway interactions; Patients; Phase; Phosphoproteins; Phosphorylation; Protein-Serine-Threonine Kinases; Proteins; Proteomics; Relapse; Research; Research Personnel; Resistance; Role; Serine; Signal Transduction; Site; System; Therapeutic; Toxic effect; Training; Transcript; United States; Variant; cancer cell; cancer therapy; cell growth; clinical efficacy; design; early phase clinical trial; effective therapy; experimental analysis; exportin 1 protein; genetic analysis; improved; in vivo; inhibitor; loss of function; mutant; new therapeutic target; novel therapeutic intervention; pancreatic ductal adenocarcinoma cell; pancreatic ductal adenocarcinoma model; pharmacologic; phosphoproteomics; post-doctoral training; preclinical development; programs; receptor; relapse patients; research clinical testing; resistance mechanism; response; spatiotemporal; standard of care; success; targeted treatment; therapeutic target; therapy development; transcriptome; tumor; tumorigenesis

Project Summary/Abstract Pancreatic ductal adenocarcinoma (PDAC), the third leading cause of cancer deaths in the United States, is characterized by a 95% rate of mutational activation of the KRAS oncogene. After nearly four decades of failure, the recent clinical approval of a direct KRAS inhibitor targeting one KRAS mutation (G12C) for lung cancer marks a significant milestone in the development of therapies for KRAS-mutant cancers. KRASG12C-specific inhibitors have demonstrated dramatic tumor shrinkage in a subset of KRASG12C-mutant patients but essentially all relapse due to treatment-induced acquired resistance. Genetic analyses of relapsed patients have identified mechanisms of resistance, with a majority involving mutational activation of signaling components that drive reactivation of the key KRAS effector pathway, the three-tiered RAF-MEK-ERK mitogen-activated protein kinase cascade. Thus, ERK reactivation will limit the long-term efficacy of direct KRAS inhibitors. Despite the highly successful development of potent and selective inhibitors of each node of the ERK MAPK cascade, when used as monotherapy, they have shown little to no clinical efficacy against RAS-mutant cancers. Two key issues have contributed to this outcome, toxicity for normal tissues and de novo or treatment-induced acquired resistance in cancer cells. I propose that further delineation of the mechanisms by which ERK drives KRAS-dependent cancer growth will guide the development of more effective anti-ERK therapies. However, the mechanisms by which ERK drives PDAC growth remain poorly understood. One major unresolved issue is how ERK activity in different subcellular compartments supports cancer growth. Aim 1 studies comprise my K99 phase of training where I will take two complementary approaches to gain a better understanding of the role of cytoplasmic and nuclear ERK activity in supporting KRAS-dependent PDAC growth. First, I will determine the capacity of cytoplasmic versus nuclear ERK activity in supporting the growth of KRAS-mutant PDAC. Second, I will use a pharmacological inhibitor of the nuclear export protein exportin-1 (Selinexor) to determine whether it disrupts ERK cytoplasmic- nuclear dynamics and sensitizes PDAC models to KRAS inhibition. My Aim 2 studies comprise my R00- supported independent research and are based on our comprehensive ERK-dependent phosphoproteome/ transcriptome studies in KRAS-mutant PDAC. Using these data, I designed a CRISPR-Cas9 genetic loss-of- function screen library, targeting ERK regulated phosphoproteins and/or transcripts. I will now perform a system- wide determination of how ERK contributes to PDAC tumorigenesis as well as identify new ERK dependent targets to combine with KRAS inhibitors. To help me achieve these research goals and successfully transition to the independent phase I have an exceptional mentoring committee comprised of leading researchers in the study of KRAS signaling and therapeutics (Channing Der), in ERK spatiotemporal signaling (Jin Zhang), and ERK substrate utilization (John Blenis). With their guidance I am confident that I will successfully transition to establish an independent research program where I will advance our knowledge on ERK signaling and therapeutics.

项目摘要/摘要 胰腺导管腺癌（PDAC）是美国癌症死亡的第三主要原因，是 特征是Kras癌基因的突变激活率为95％。经过将近四十年的失败， 靶向一个KRAS突变（G12C）的直接KRAS抑制剂的最新临床批准用于肺癌标记 Kras突变癌的疗法开发的重要里程碑。 KRASG12C特异性抑制剂 在KRASG12C突变患者的一部分中表现出了戏剧性的肿瘤收缩，但本质上都是复发 由于治疗引起的获得性抗性。复发患者的遗传分析已鉴定出机制 电阻，大多数涉及信号成分的突变激活，以驱动驱动的重新激活 关键的KRAS效应途径，三层RAF-MEK-MEK-MEK-MEK-ERK有丝分裂原激活的蛋白激酶级联反应。 因此，ERK重新激活将限制直接KRAS抑制剂的长期疗效。尽管非常成功 当用作ERK MAPK CASCADE每个节点的有效和选择性抑制剂的发展 单一疗法，它们对RAS突变癌的临床功效几乎没有。两个关键问题有 有助于这种结果，对正常组织的毒性和从头开始或治疗引起的获得的耐药性 癌细胞。我建议进一步描述ERK驱动Kras依赖性癌症的机制 增长将指导更有效的抗ERK疗法的发展。但是， ERK驱动PDAC的增长仍然知之甚少。一个未解决的主要问题是ERK如何在不同的 亚细胞室支持癌症的生长。 AIM 1研究包括我的K99培训阶段 采用两种补充方法，以更好地了解细胞质和核ERK的作用 支持KRAS依赖性PDAC生长的活性。首先，我将确定细胞质与 核ERK活性在支持KRAS突变PDAC的生长方面。第二，我将使用药理 核输出蛋白Exportin-1（selinexor）的抑制剂，以确定它是否破坏ERK细胞质 - 核动力学并将PDAC模型敏感到KRAS抑制。我的目标2研究包括我的R00- 支持独立研究，并基于我们综合的ERK依赖性磷蛋白组/ KRAS突变PDAC中的转录组研究。使用这些数据，我设计了CRISPR-CAS9遗传丧失 - 功能屏幕库，针对ERK调节的磷蛋白和/或转录本。我现在将执行一个系统 - 广泛确定ERK如何促进PDAC肿瘤发生，并确定依赖新ERK 与KRAS抑制剂结合的靶标。为了帮助我实现这些研究目标，并成功过渡到 独立阶段I有一个杰出的指导委员会，由研究中的主要研究人员组成 ERK时空信号（Jin Zhang）和ERK中的KRAS信号传导和治疗学（Channing der）的 底物利用率（John Blenis）。在他们的指导下，我有信心我将成功过渡以建立 一个独立的研究计划，我将提高我们对ERK信号和治疗学的知识。