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.

项目概要 胃肠道间质瘤 (GIST) 治疗的管理发生了革命性的变化 KIT和PDGFRA的激活突变及晚期酪氨酸激酶抑制剂（TKI）的临床应用 疾病。然而，对所有已批准的 TKI 系的耐药性是转移性癌症中危及生命的临床障碍。 GIST 患者，患有由 KIT/PDGFRA 中多种耐药突变组成的多克隆肿瘤。 开发难治性胃肠道间质瘤的有效治疗策略需要确定新的靶点。使用 通过激酶组分析和功能丧失检测，我们发现 GIST 高度依赖于 Wee1 活性。 Wee1 长期以来被认为是控制基因组完整性的关键 G2/M 细胞周期检查点看门人 调节增殖。然而，最近的研究表明 Wee1 直接或间接地与其他 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 GIST 细胞系的分析强调了致癌 KIT/PDGFRA 在 DNA 损伤反应中的作用 （DDR）和复制压力。我们的体外和体内研究揭示了 adavosertib (Wee1 抑制剂）与 avapritinib（KIT/PDGFRA 抑制剂）联合治疗 TKI 敏感的 KIT 和 PDGFRA 突变 GIST 系和患者来源的异种移植（PDX）模型中。此外，我们在 TKI 难治性方面的初步数据 设置证明了 Wee1 抑制和拓扑异构酶 II 抑制剂之间的治疗协同作用， 阿霉素，在一组 TKI 抗性 GIST 细胞系中处于低纳摩尔浓度，表明 Wee1 抑制作用使难治性 GIST 对阿霉素敏感，提供了 KIT 和 PDGFRA 之外的可行靶点。我们 假设致癌 KIT/PDGFRA 激活 DDR 蛋白以抑制复制应激和 DNA 损伤 维持 GIST 基因组稳定性，从而对 DNA 损伤剂产生抵抗力。我们的初步调查结果 提供了一个强有力的前提，即 Wee1 是该过程的关键调节者，并且其抑制作用具有潜力 胃肠道间质瘤（GIST）肿瘤的治疗。研究 Wee1 和 KIT/PDGFRA 协同性的机制 GIST 对 DNA 损伤和复制应激的反应有必要了解这一关键的细胞保护作用 GIST 中的机制，这可能代表了一种新的治疗脆弱性。最终，该提案旨在 检验 Wee1 抑制将成为目前治疗 GIST 患者的一个有价值的补充的假设 TKI 敏感或耐药疾病。我们相信本提案中概述的研究将提供新颖的见解 深入了解 Wee1 在维持基因组完整性以及支持 GIST 临床试验的临床前数据方面的作用。