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概要/摘要 在过去的三十年里，上皮性卵巢癌患者的5年生存率有所提高，但长期以来， 长期生存率没有变化，总体保持在约30%。在手术细胞减少后，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。 单用铂类化合物的初级治疗在约70%的卵巢癌中产生消退。 而42%的人对紫杉醇单独治疗有反应，两种药物之间没有协同作用。我们长久以来- 长期翻译的目标是增加卵巢癌化疗的有效性。虽然许多 研究人员一直专注于克服对紫杉烷类的获得性耐药性， 在初次化疗期间给予紫杉醇以增强紫杉醇反应。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（SIK 2）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中。SIK 2是正常中心体分裂所必需的，并且在34%的人中过表达。 所有组织类型的卵巢癌，与总生存率下降相关。在有丝分裂过程中， 自磷酸化和磷酸化C-Nap 1，一种介导中心体分裂的结构蛋白。SIK2 还磷酸化p85α，驱动PI 3 K和HDAC 5的活化，修饰染色质和DNA修复。 我们与Arrien Pharmaceuticals建立了合作关系，开发口服小剂量 SIK 2的分子量抑制剂：ARN-3236和ARN-3261，其不同之处在于单一溶剂结合 替代。ARN-3236抑制10种卵巢癌细胞系的生长，IC 50为0.8 - 2.6 μM，其中 ARN-3236的IC 50与内源性SIK 2表达呈负相关（Pearson's r =-0.642，P = 0.03）。 ARN-3236还在10个细胞系中的8个以及SKOv 3 ip中增强了对紫杉醇的敏感性（P = 0.028）， OVCAR 8异种移植物。ARN-3236在间期使中心体与细胞核解偶联， 有丝分裂中的中心体分离，引起前中期停滞和诱导凋亡细胞死亡，以及 四倍性ARN-3236还抑制AKT磷酸化并减弱存活素表达。ARN-3261 在异种移植模型中增强了对紫杉醇和顺铂的敏感性，并表现出很小的毒性，以及 对PgP的抗性更强，效力是ARN-3236的13倍。我们将追求三个目标：1）执行 SIK 2抑制剂ARN-3261单独使用（IA）和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（SIK 2）和药效学（pSIK 2和pSIK 3）。 pHDAC 5）生物标志物，以及多倍性、pAKT、存活素和细胞凋亡的生物标志物的水平; 2） 确定SIK 2抑制剂ARN-3261是否增强对卡铂/紫杉醇的反应， 卡铂/脂质体多柔比星在卵巢癌细胞系来源的和患者来源的异种移植物中的作用;和 鉴定在用SIK 2抑制剂处理的卵巢癌细胞中产生合成致死性的靶点。