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.

项目总结/摘要 胰腺导管腺癌（PDAC）是美国癌症死亡的第三大原因， 其特征在于95%的KRAS癌基因突变激活率。经过近四十年的失败， 最近临床上批准了一种针对肺癌标志物的KRAS突变（G12C）的直接KRAS抑制剂， 这是KRAS突变型癌症治疗发展的重要里程碑。KRASG12C特异性抑制剂 在KRASG12C突变患者的一个亚组中显示出显著的肿瘤缩小，但基本上所有的复发 由于治疗诱导的获得性耐药性。对复发患者的基因分析已经确定了机制 大多数涉及信号成分的突变激活，这些信号成分驱动抗性的重新激活。 关键的KRAS效应通路，三层RAF-MEK-ERK促分裂原活化蛋白激酶级联。 因此，ERK再激活将限制直接KRAS抑制剂的长期疗效。尽管非常成功， 开发ERK MAPK级联的每个节点的有效和选择性抑制剂，当用作 然而，在单一疗法中，它们对RAS突变型癌症显示出很少或没有临床功效。有两个关键问题 导致这一结果，正常组织的毒性和新生或治疗诱导的获得性耐药， 癌细胞我建议进一步阐明ERK驱动KRAS依赖性癌症的机制 生长将指导更有效的抗ERK疗法的发展。然而， ERK驱动PDAC生长仍然知之甚少。一个尚未解决的主要问题是ERK活性如何在不同的细胞中表达。 亚细胞区室支持癌症生长。目标1研究包括我的K99培训阶段，我将 采取两种互补的方法来更好地了解细胞质和细胞核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活性支持KRAS突变体PDAC的生长。第二，我会用一种药物 核输出蛋白exportin-1（Selinexor）的抑制剂，以确定其是否破坏ERK细胞质- 核动力学，并使PDAC模型对KRAS抑制敏感。我的目标2研究包括我的R00- 支持独立研究，并基于我们全面的ERK依赖磷酸蛋白质组/ 在KRAS突变体PDAC中的转录组研究。利用这些数据，我设计了一个CRISPR-Cas9基因缺失- 功能筛选文库，靶向ERK调节的磷蛋白和/或转录物。我要表演一个系统- 广泛确定ERK如何促进PDAC肿瘤发生以及鉴定新的ERK依赖性 靶点与KRAS抑制剂联合收割机。帮助我实现这些研究目标，并成功地过渡到 独立的第一阶段有一个特殊的指导委员会，由研究中的主要研究人员组成 KRAS信号传导和治疗（Channing Der）、ERK时空信号传导（Jin Zhang）和ERK 底物利用（John Blenis）。在他们的指导下，我相信我将成功地过渡到建立 这是一个独立的研究项目，我将在其中推进我们对ERK信号传导和治疗学的知识。