Elucidating the critical role of Wee1 in GIST

阐明 Wee1 在 GIST 中的关键作用

• 批准号: 10681775
• 负责人: Lori Rink
• 金额: $ 43.01万
• 依托单位: RESEARCH INST OF FOX CHASE CAN CTR
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-03-01 至 2028-02-29
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10681775
• 关键词: Acute Myelocytic Leukemia; Biological Assay; Cell Cycle; Cell Cycle Checkpoint; Cell Survival; Cell physiology; Cells; Clinical; Clinical Research; Clinical Trials; Combined Modality Therapy; DNA; DNA Damage; DNA Repair; DNA biosynthesis; DNA damage checkpoint; DNA replication fork; Data; Development; Disease Resistance; Down-Regulation; Doxorubicin; Evaluation; Fiber; Future; G2/M Checkpoint Pathway; Gastrointestinal Stromal Tumors; Gatekeeping; Gene set enrichment analysis; Genes; Genome Stability; Genomic Instability; Goals; Hyperactivity; Imatinib mesylate; Immunofluorescence Microscopy; In Vitro; Left; Libraries; Life; Malignant Neoplasms; Mediator; Mutagens; Mutate; Mutation; Oncogenic; PDGFRA gene; PDGFRB gene; Pathway interactions; Patients; Phosphotransferases; Process; Proliferating; Proteins; Refractory; Regulation; Resistance; Role; Serine; Signal Transduction; Specimen; Stress; Technology; Testing; Therapeutic; Threonine; Time; Topoisomerase-II Inhibitor; Tumor Cell Line; Tyrosine Kinase Inhibitor; Xenograft Model; advanced disease; avapritinib; cancer genome; chemotherapy; chromatin remodeling; clinical application; effective therapy; genome integrity; genotoxicity; homologous recombination; improved; in vivo; ineffective therapies; inhibitor; innovative technologies; insight; loss of function; mutant; nanomolar; neoplastic cell; new therapeutic target; novel; novel therapeutics; patient derived xenograft model; pre-clinical; prevent; receptor; replication stress; resistance mutation; response; restoration; single molecule; success; synergism; therapy resistant; transcriptome sequencing; transcriptomic profiling; treatment strategy; tumor; tumor progression

Project Summary Management of gastrointestinal stromal tumor (GIST) treatment has been revolutionized by the identification of activating mutations in KIT and PDGFRA and clinical application of tyrosine kinase inhibitors (TKIs) in advanced disease. However, resistance to all approved lines of TKIs is a life-threatening clinical obstacle in metastatic GIST patients, who have polyclonal tumors comprised of a variety of resistance mutations in KIT/PDGFRA. Development of effective treatment strategies for refractory GIST requires identification of novel targets. Using kinome profiling and loss-of function assays, we found that GIST are exquisitely dependent on Wee1 activity. Wee1 has long been known as a critical G2/M cell cycle checkpoint gatekeeper controlling genomic integrity and regulating proliferation. However, recent studies have implicated Wee1, both directly and indirectly, in other cellular functions, including restoration of stalled replication forks and chromatin remodeling. Transcriptome profiling of TKI-sensitive GIST cell lines highlighted a role for oncogenic KIT/PDGFRA in DNA damage response (DDR) and replication stress. Our in vitro and in vivo studies revealed significant efficacy of adavosertib (Wee1 inhibitor) in combination with avapritinib (KIT/PDGFRA inhibitor) in TKI-sensitive KIT and PDGFRA-mutant GIST lines and in a patient-derived xenograft (PDX) model. Furthermore, our preliminary data in the TKI-refractory setting demonstrated therapeutic synergies between Wee1 inhibition and the topoisomerase II inhibitor, doxorubicin, at low nanomolar concentrations in a panel of TKI-resistant GIST cell lines, suggesting Wee1 inhibition sensitizes refractory GIST to doxorubicin, providing viable targets beyond KIT and PDGFRA. We hypothesize that oncogenic KIT/PDGFRA activates DDR proteins to inhibit replicative stress and DNA damage and maintain genomic stability in GIST leading to resistance to DNA damaging agents. Our preliminary findings provide a strong premise that Wee1 is a critical mediator of this process and that its inhibition has potential in the treatment of GIST tumors. Studies interrogating mechanisms of Wee1 and KIT/PDGFRA cooperativity in response to DNA damage and replication stress in GIST are warranted to understand this critical cell-protective mechanism in GIST, which may represent a novel therapeutic vulnerability. Ultimately, this proposal seeks to test the hypothesis that Wee1 inhibition will be a valuable addition to current treatments for GIST patients with TKI-sensitive or -resistant disease. We believe that the studies outlined in this proposal will provide novel insights into the role of Wee1 in maintaining genomic integrity as well as preclinical data to support clinical trials in GIST.

项目摘要 胃肠道肿瘤（GIST）治疗的治疗已通过鉴定已彻底改变 在盒和PDGFRA中激活突变以及酪氨酸激酶抑制剂（TKI）在晚期中的临床应用 疾病。但是，对所有批准的TKI线的抵抗是威胁生命的转移性临床障碍 GIST患者，患有多克隆肿瘤，由KIT/PDGFRA中的各种耐药突变组成。 制定有效的耐火性要点的治疗策略需要确定新目标。使用 Kinome分析和功能丧失测定法，我们发现要点非常取决于WEE1活动。 WEE1长期以来一直被称为临界G2/M细胞周期检查点守门人控制基因组完整性和 调节增殖。但是，最近的研究已直接和间接地参与了其他研究 细胞功能，包括恢复停滞的复制叉和染色质重塑。转录组 TKI敏感的GIST细胞系的分析强调了致癌试剂盒/PDGFRA在DNA损伤响应中的作用 （DDR）和复制应力。我们的体外和体内研究揭示了adavosertib的显着疗效（WEE1 抑制剂）与TKI敏感试剂盒中的Avapritinib（Kit/PDGFRA抑制剂）结合使用 线和患者衍生的异种移植（PDX）模型。此外，我们的初步数据 设置表明WEE1抑制与拓扑异构酶II抑制剂之间的治疗协同作用， 阿霉素，在一系列TKI抗性的GIST细胞系中低纳摩尔浓度，表明WEE1 抑制作用使难治性的角度对阿霉素敏感，从而超出了Kit和PDGFRA。我们 假设致癌试剂盒/PDGFRA激活DDR蛋白以抑制复制应力和DNA损伤 并保持要点的基因组稳定性，从而导致对DNA损害剂的抗性。我们的初步发现 提供了一个有力的前提，即WEE1是该过程的关键调解人，其抑制作用具有潜力 要点肿瘤的治疗。研究询问WEE1和KIT/PDGFRA合作的机制 有必要理解对GIST中对DNA损伤和复制应力的反应，以了解这种关键的细胞保护 要点机制，这可能代表了一种新型的治疗脆弱性。最终，该提议试图 检验WEE1抑制作用的假设将是目前针对要点患者的当前治疗 TKI敏感或抗性疾病。我们认为，该提案中概述的研究将提供新颖的见解 WEE1在维持基因组完整性以及临床前数据中的作用，以支持GIST的临床试验。