Project 4: SIK2 PROVIDES A NOVEL TARGET FOR OVARIAN CANCER THERAPY IN COMBINATION WITH PACLITAXEL AND INHIBITORS OF PARP

项目 4：SIK2 结合紫杉醇和 PARP 抑制剂为卵巢癌治疗提供新靶点

• 批准号: 10005298
• 负责人: ROBERT C BAST
• 金额: $ 32.9万
• 依托单位: UNIVERSITY OF TX MD ANDERSON CAN CTR
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-22 至 2022-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10005298
• 关键词: Apoptosis; Apoptotic; Attention; Attenuated; Automobile Driving; Binding; Biological Assay; Biological Markers; Biopsy; Blood; Cancer Center; Cancer cell line; Carboplatin; Cell Death; Cell Line; Cell Nucleus; Cells; Centrosome; Chemicals; Chromatin; Cisplatin; Collaborations; DNA Repair; Data; Doctor of Medicine; Dose; Drug Combinations; Drug Kinetics; Drug resistance; Effectiveness; Enzymes; Epithelial ovarian cancer; Exhibits; Goals; Growth; HDAC5 gene; Interphase; Leukocytes; Liposomal Doxorubicin; Malignant Neoplasms; Malignant neoplasm of ovary; Measures; Mediating; Mitosis; Molecular Weight; Oral; Paclitaxel; Patients; Peripheral Blood Mononuclear Cell; Pharmaceutical Preparations; Pharmacodynamics; Pharmacologic Substance; Phosphorylation; Phosphotransferases; Platinum; Polyploidy; Prometaphase; Protein-Serine-Threonine Kinases; Proteins; Proto-Oncogene Proteins c-akt; Research Personnel; Resistance; Signal Transduction; Signaling Protein; Small Interfering RNA; Sodium Chloride; Solvents; Structural Protein; Testing; Tetraploidy; Toxic effect; Tumor Debulking; Woman; Xenograft Model; Xenograft procedure; base; cancer cell; cancer therapy; chemotherapy; improved; inhibitor/antagonist; knock-down; neoplastic cell; novel; overexpression; pharmacodynamic biomarker; phase I trial; predicting response; predictive marker; resistance mechanism; response; standard care; survivin; synergism; taxane; three dimensional cell culture; tumor

Project 4 SUMMARY/ABSTRACT Over the last three decades, 5-year survival has improved for patients with epithelial ovarian cancer, but long- term survival has not changed and remains at approximately 30% overall. Following surgical cytoreduction, all new patients receive standard primary chemotherapy that includes a combination of carboplatin and paclitaxel. Primary therapy with platinum-based compounds alone produces regression in approximately 70% of ovarian cancers, whereas 42% respond to paclitaxel alone and there is no synergy between the two drugs. Our long- term translational goal is to increase the effectiveness of chemotherapy for ovarian cancer. While many investigators have focused on overcoming acquired resistance to taxanes, relatively little attention has been given to enhancing paclitaxel response during primary chemotherapy. Using a functional siRNA screen for kinases that regulate sensitivity to paclitaxel, we found that knockdown of the serine-threonine kinase salt- induced kinase 2 (SIK2) induces polyploidy, inhibits ovarian cancer growth and enhances paclitaxel sensitivity in cell lines and xenografts. SIK2 is required for normal centrosome splitting and is overexpressed in 34% of ovarian cancers of all histotypes, associated with decreased overall survival. During mitosis, SIK2 undergoes autophosphorylation and phosphorylates C-Nap1 a structural protein that mediates centrosome splitting. SIK2 also phosphorylates p85α, driving activation of PI3K, as well as HDAC5, modifying chromatin and DNA repair. We have established a collaboration with Arrien Pharmaceuticals to develop orally administered small molecular weight inhibitors of SIK2: ARN-3236 and ARN-3261 that differ by a single solvent binding substitution. ARN-3236 inhibited growth of 10 ovarian cancer cell lines at an IC50 of 0.8 to 2.6 μM, where the IC50 of ARN-3236 was inversely correlated with endogenous SIK2 expression (Pearson’s r = -0.642, P = 0.03). ARN-3236 also enhanced sensitivity to paclitaxel in 8 of 10 cell lines, as well as in SKOv3ip (P = 0.028) and OVCAR8 xenografts. ARN-3236 uncoupled the centrosome from the nucleus in interphase, blocked centrosome separation in mitosis, caused prometaphase arrest and induced apoptotic cell death as well as tetraploidy. ARN-3236 also inhibited AKT phosphorylation and attenuated survivin expression. ARN-3261 enhanced sensitivity to paclitaxel and cisplatin in xenograft models and exhibited little toxicity, as well as greater resistance to PgP and13-fold greater potency than ARN-3236. We will pursue three aims: 1) to perform a phase I trial of the SIK2 inhibitor ARN-3261 alone (IA) and in combination with weekly paclitaxel (IB) measuring pharmacokinetics of ARN-3261 and paclitaxel, predictive (SIK2) and pharmacodynamic (pSIK2 and pHDAC5) biomarkers, as well as levels of polyploidy, pAKT, survivin and biomarkers for apoptosis; 2) to determine whether the SIK2 inhibitor ARN-3261 enhances response to carboplatin/paclitaxel and carboplatin/liposomal doxorubicin in ovarian cancer cell line-derived and patient-derived xenografts; and 3) to identify targets that produce synthetic lethality in ovarian cancer cells treated with SIK2 inhibitors.

项目 4 摘要/摘要 在过去三十年中，上皮性卵巢癌患者的 5 年生存率有所提高，但长期 长期生存率没有变化，总体保持在 30% 左右。手术细胞减灭术后，所有 新患者接受标准的主要化疗，包括卡铂和紫杉醇的组合。 仅使用铂类化合物进行初级治疗可使约 70% 的卵巢功能消退 癌症，而 42% 对单独的紫杉醇有反应，并且两种药物之间没有协同作用。我们的长期 术语转化目标是提高卵巢癌化疗的有效性。虽然很多 研究人员将重点放在克服对紫杉烷类药物的获得性耐药性上，但相对较少的关注 用于增强初次化疗期间紫杉醇的反应。使用功能性 siRNA 筛选 调节对紫杉醇敏感性的激酶，我们发现丝氨酸-苏氨酸激酶盐的敲低- 诱导激酶 2 (SIK2) 诱导多倍体、抑制卵巢癌生长并增强紫杉醇敏感性 在细胞系和异种移植物中。 SIK2 是正常中心体分裂所必需的，并且在 34% 的细胞中过表达 所有组织型的卵巢癌，与总生存率降低相关。在有丝分裂期间，SIK2 经历 自磷酸化和磷酸化 C-Nap1（一种介导中心体分裂的结构蛋白）。 SIK2 它还磷酸化 p85α，驱动 PI3K 和 HDAC5 的激活，从而修改染色质和 DNA 修复。 我们与 Arrien Pharmaceuticals 建立了合作，开发口服小剂量 SIK2 的分子量抑制剂：ARN-3236 和 ARN-3261，其区别在于单一溶剂结合 替代。 ARN-3236 抑制 10 种卵巢癌细胞系的生长，IC50 为 0.8 至 2.6 μM，其中 ARN-3236 的 IC50 与内源性 SIK2 表达呈负相关（Pearson’s r = -0.642，P = 0.03）。 ARN-3236 还增强了 10 个细胞系中的 8 个以及 SKOv3ip 中对紫杉醇的敏感性 (P = 0.028) 和 OVCAR8 异种移植物。 ARN-3236 在间期将中心体与细胞核解偶联，阻断 有丝分裂中的中心体分离，导致中期停滞并诱导细胞凋亡 四倍体。 ARN-3236 还抑制 AKT 磷酸化并减弱生存素表达。 ARN-3261 在异种移植模型中增强对紫杉醇和顺铂的敏感性，并且毒性很小，并且 比 ARN-3236 具有更强的 PgP 耐受性和 13 倍的效力。我们将追求三个目标：1） SIK2 抑制剂 ARN-3261 单独使用 (IA) 以及与每周紫杉醇联合使用 (IB) 的 I 期试验 测量 ARN-3261 和紫杉醇的药代动力学、预测 (SIK2) 和药效学 (pSIK2 和 pHDAC5) 生物标志物，以及多倍体、pAKT、生存素和细胞凋亡生物标志物的水平； 2）到 确定 SIK2 抑制剂 ARN-3261 是否增强对卡铂/紫杉醇的反应以及 卡铂/脂质体阿霉素在卵巢癌细胞系来源和患者来源的异种移植物中的作用； 3) 至 确定在用 SIK2 抑制剂治疗的卵巢癌细胞中产生合成致死作用的靶标。