Clinical Translation of Nuclear Export Inhibitor in Metastatic Pancreatic Cancer

核输出抑制剂在转移性胰腺癌中的临床转化

• 批准号: 10083197
• 负责人: Asfar S Azmi
• 金额: $ 35.23万
• 依托单位: WAYNE STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-01-15 至 2022-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10083197
• 关键词: Biological; Biopsy; CRISPR/Cas technology; Cell Death; Cell Death Signaling Process; Cell Maintenance; Cell Nucleus; Cell model; Cells; Clinical; Clinical Research; Computational Biology; Correlative Study; Coupled; Critical Pathways; Data; Desmoplastic; Diagnosis; Disease; Expression Profiling; FBXW7 gene; Fibroblasts; Gene Mutation; Genes; Global Change; Growth; In Vitro; Inhibition of Apoptosis; KPC model; Knowledge; Malignant neoplasm of pancreas; Methodology; Modality; Modeling; Molecular Biology; Nuclear; Nuclear Export; Nuclear Proteins; Oral; Organoids; Outcome; PAWR gene; Paclitaxel; Pancreatic Ductal Adenocarcinoma; Pathway Analysis; Pathway interactions; Patient Recruitments; Patients; Penetrance; Pharmaceutical Preparations; Phase; Phase I Clinical Trials; Phase II Clinical Trials; Phase Ib/II Clinical Trial; Phase Ib/II Trial; Pre-Clinical Model; Process; Proteins; RNA Interference; Recruitment Activity; Regimen; Reporting; Residual Tumors; Resistance; Role; Safety; Signal Transduction; Specificity; Specimen; Time; Transgenic Mice; Transgenic Model; Transgenic Organisms; Tumor Suppressor Proteins; Vascularization; Work; analog; biomarker development; cancer stem cell; chemotherapeutic agent; clinical translation; cytotoxic; epithelial to mesenchymal transition; exportin 1 protein; gemcitabine; genome editing; improved; in vivo; inhibitor/antagonist; innovation; model building; mouse model; novel therapeutics; overexpression; pancreatic ductal adenocarcinoma cell; pancreatic ductal adenocarcinoma model; pharmacodynamic biomarker; pre-clinical; preclinical study; response biomarker; stem cell growth; stemness; subcutaneous; synergism; therapeutic biomarker; therapy design; therapy outcome; transcriptome sequencing; treatment response; tumor

Summary: Pancreatic ductal adenocarcinoma (PDAC) remains a deadly disease in urgent need of newer therapeutic modalities. PDAC tumors are very heterogeneous carrying alterations in many critical pathways and require a broad form of therapy that can activate multiple tumor suppressor proteins (TSPs) simultaneously. We have shown that in PDAC over-expression of the nuclear exporter protein Exportin 1 (XPO1) leads to functional inactivation of TSPs (such as FOXO3a, p27, Par-4) through their mislocalization. We were the first to demonstrate that inhibition of XPO1 by CRISPR/Cas9 validated Selective Inhibitor of Nuclear Export (SINE) selinexor and analogs can restore the anti-tumor function of multiple TSPs leading to PDAC cell death and tumor inhibition in orthotopic models. More significantly, our findings show that SINE synergize with gemcitabine (GEM) and nab-paclitaxel leading to enhanced PDAC growth inhibition, apoptosis, reversal of epithelial-to-mesenchymal transition (EMT) and spheroid disintegration of PDAC derived cancer stem cells (CSCs). The observed synergy was due in part to enhanced nuclear localization of TSPs and suppression of CSC and EMT markers. Pathway analysis showed enhancement in cell death signaling and suppression of CSC sustaining networks. Most importantly we observed activation of fibroblast specific cell death pathways highlighting an as of yet unexplored role of XPO1 in sustaining fibroblast growth that is recognized to support high desmoplastic reaction (DR) in PDAC. Our multi-model pre-clinical work has led to a Phase Ib/II clinical study involving GEM-nab-paclitaxel-selinexor for metastatic PDAC (NCT02178436). In this proposal we hypothesize that XPO1 inhibition will enforce the nuclear retention of TSPs, restore their anti- tumor function leading to enhancement of cytotoxic efficacy of GEM-nab-paclitaxel in PDAC. To validate our hypothesis, we propose to integrate high-throughput RNA-Seq and SILAC-MS methodologies in primary cellular and transgenic mice models and in a Phase Ib/II trial combining selinexor with Gem-nab-paclitaxel in PDAC patients. We anticipate that our holistic analyses will, for the first time, delineate the impact of XPO1 inhibition in re-programming of PDAC CSC and DR networks that leads to the enhancement in the efficacy of GEM-nab-paclitaxel in tumors. In Aim1 we will demonstrate in vitro synergy between GEM-nab-paclitaxel and selinexor in primary PDAC models using high-throughput methodologies. In Aim2 we will demonstrate the efficacy and synergy of GEM-nab-paclitaxel-selinexor in LSL-KrasG12D/+;LSL-Trp53R172H/+;Pdx-1-Cre mice model. These studies will identify sensitivity and/or resistance markers that will support our phase Ib/II trial in Aim3 in which we intend to determine the safety and efficacy of selinexor when combined with GEM-nab- paclitaxel in patients with metastatic PDAC. Biopsies from this trial will be used to validate locked pharmacodynamic markers of selinexor target engagement in patients. Clinical Impact: The successful completion our studies will bring forward a new therapy to improve the survival of patients with PDAC.

摘要：胰腺导管腺癌（PDAC）仍然是一种致命的疾病，迫切需要更新 治疗方式。 PDAC肿瘤在许多关键途径中是非常异构的携带变化 并需要一种可以激活多种肿瘤抑制蛋白（TSP）的广泛的治疗形式 同时地。我们已经表明，在PDAC中，核出口蛋白导出1的过表达1 （XPO1）导致TSP的功能失活（例如FOXO3A，P27，PAR-4）通过其错误定位。 我们是第一个证明CRISPR/CAS9抑制XPO1的人 核出口（SINE）selinexor和类似物可以恢复多个TSP的抗肿瘤功能 PDAC细胞死亡和原位模型中的肿瘤抑制。更重要的是，我们的发现表明正弦 与吉西他滨（GEM）和NAB-甲酰胺协同作用，导致PDAC生长抑制，凋亡，凋亡， PDAC衍生癌的上皮到间质转变（EMT）和球体分解的逆转 干细胞（CSC）。观察到的协同作用部分是由于TSP和 CSC和EMT标记的抑制。途径分析显示细胞死亡信号传导和 抑制CSC维持网络。最重要的是，我们观察到成纤维细胞的激活 死亡途径突出显示XPO1在维持成纤维细胞生长中的作用尚未开发 公认的PDAC中支持高脱糖反应（DR）。我们的多模型临床前工作导致了 IB/II期临床研究涉及转移性PDAC（NCT02178436）的GEM-NAB-链酰甲酰基轴线。在这个 提案我们假设XPO1抑制作用将强制执行TSP的核保留率，恢复其抗 肿瘤功能可增强PDAC中宝石-NAB-甲酰胺的细胞毒性功效。验证我们的 假设，我们建议将高通量RNA-seq和SILAC-MS方法整合在原发性 细胞和转基因小鼠模型，以及在IB/II期试验中，将selinexor与宝石 - 纳布 - 巴列赛相结合 PDAC患者。我们预计我们的整体分析将首次描述XPO1的影响 抑制PDAC CSC和DR网络的重新编程，这些网络导致增强的功效 肿瘤中的宝石 - nab-甲酰胺。在AIM1中，我们将在Gem-Nab-Paclitaxel和 使用高通量方法的主要PDAC模型中的selinexor。在AIM2中，我们将证明 LSL-KRASG12D/+; LSL-TRP53R172H/+; PDX-1-CRE小鼠的gem-nab-paclitaxel-selinexor的效果和协同效果 模型。这些研究将确定灵敏度和/或阻力标记，这些标记将支持我们的IB/II期试验 我们打算在其中确定selinexor的安全性和功效的AIM3与gem-nab-相结合 转移性PDAC患者的紫杉醇。该试验的活检将用于验证锁定 Selinexor靶标参与患者的药效学标记。临床影响：成功 完成我们的研究将提出一种新的疗法，以改善PDAC患者的生存。