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: 10321222
• 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/10321222
• 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; 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.

胰腺导管腺癌（PDAC）仍然是一种致命的疾病，迫切需要新的治疗方法。 治疗方式PDAC肿瘤是非常异质性的，在许多关键途径中携带改变 需要一种广泛的治疗方法，可以激活多种肿瘤抑制蛋白（TSP） 同步我们已经证明，在PDAC中，核输出蛋白Exportin 1的过表达 （XPO 1）通过其错误定位导致TSP（如FOXO 3a、p27、Par-4）的功能失活。 我们是第一个证明CRISPR/Cas9对XPO 1的抑制验证了XPO 1的选择性抑制剂。 核输出（SINE）赛灵克斯和类似物可以恢复多种TSP的抗肿瘤功能， 原位模型中的PDAC细胞死亡和肿瘤抑制。更重要的是，我们的研究结果表明，SINE 与吉西他滨（GEM）和白蛋白结合型紫杉醇协同作用，导致PDAC生长抑制、细胞凋亡 PDAC衍生的癌的上皮-间质转化（EMT）和球体崩解的逆转 干细胞（CSCs）。观察到的协同作用部分是由于TSP的核定位增强， 抑制CSC和EMT标志物。通路分析显示细胞死亡信号传导增强， 抑制CSC维持网络。最重要的是，我们观察到成纤维细胞特异性细胞的活化， 死亡途径强调了XPO 1在维持成纤维细胞生长中尚未探索的作用， 被认为支持PDAC中的高促纤维增生反应（DR）。我们的多模型临床前工作已经导致了 涉及GEM-nab-紫杉醇-赛林克斯用于转移性PDAC的Ib/II期临床研究（NCT 02178436）。在这 我们假设XPO 1抑制将加强TSP的核滞留，恢复它们的抗- 肿瘤功能，导致GEM-nab-紫杉醇在PDAC中的细胞毒性功效增强。来验证我们 假设，我们建议将高通量RNA-Seq和SILAC-MS方法集成在初级 细胞和转基因小鼠模型，并在Ib/II期试验中联合赛林克斯与Gem-nab-紫杉醇， PDAC患者我们预计，我们的整体分析将首次描述XPO 1的影响 抑制PDAC CSC和DR网络的重编程，导致增强 肿瘤中的GEM-nab-紫杉醇。在Aim 1中，我们将证明GEM-nab-紫杉醇和紫杉醇之间的体外协同作用。 使用高通量方法在主要PDAC模型中使用selinexor。在Aim 2中，我们将演示 GEM-nab-紫杉醇-司林克斯在LSL-KrasG 12 D/+; LSL-Trp 53 R172 H/+;Pdx-1-Cre小鼠中功效和协同作用 模型这些研究将确定敏感性和/或耐药标志物，以支持我们的Ib/II期试验， 目的3，我们打算确定赛林克斯与GEM-nab联合使用时的安全性和有效性。 紫杉醇治疗转移性PDAC患者。本试验的活检将用于验证锁定 患者中赛林克斯靶向接合的药效学标志物。临床影响：成功的 完成我们的研究将提出一种新的治疗方法，以提高PDAC患者的生存率。