Overcoming therapeutic resistance of gliomas

描述(申请人提供)：溶瘤单纯疱疹病毒(OHSV)-1的特定病毒基因的突变已被证明对癌细胞具有选择性，这使得在人类中进行翻译研究成为可能。然而，在第一阶段和第二阶段的临床试验中，只有一小部分接种了OHSV-1的癌症患者显示出客观的反应。我们最近已经鉴定出对OHSV介导的肿瘤分解具有抗性的GBM株系。基于我们的发现，我们创造了重组OHSV-TRAIL，一种携带可分泌的促凋亡肿瘤坏死因子相关凋亡诱导配体(TRAIL)的OHSV-TRAIL，并假设OHSV-TRAIL可用于癌症治疗，以基于机制的方式靶向广泛的耐药肿瘤。在这个方案中，我们将筛选一组从患者来源的原代GBM株分离的胶质瘤干细胞(GSC)，以检测它们对OHSV和TRAIL的敏感性，并鉴定对OHSV或OHSV和TRAIL都具有抗性的株系。根据我们对OHSV-TRAIL诱导的OHSV和TRAIL耐药胶质瘤细胞ERK表达下调、JNK和p38激活以及caspase表达上调的初步研究，我们推测OHSV将靶向于细胞增殖途径，并启动OHSV和/或TRAIL耐药的胶质瘤细胞和原代GSCs对TRAIL介导的凋亡。OHSV-S-TRAIL的有效性将在体外和由已识别的株系创建的小鼠模型中进行评估，并最终在通过筛选一组新切除的患者肿瘤而确定的耐药系中进行。将基因工程荧光(GFP和mCherry)和生物发光(萤火虫荧光素酶；Fluc和Renilla荧光素酶；Rluc)成像标记整合到OHSV和脑胶质瘤细胞中，将使我们能够通过体内生物发光成像(BLI)和活体显微镜(IVM)追踪OHSV-TRAIL的有效性。我们预计我们的发现将对以下方面做出重大贡献：1)阐明OHSV介导的肿瘤分解和TRAIL如何协同发挥作用的分子机制，以靶向对病毒溶瘤和/或细胞毒治疗具有抵抗力的异种胶质瘤细胞；以及2)最终为脑肿瘤患者开发基于OHSV的新疗法。

DESCRIPTION (provided by applicant): Mutations of specific viral genes of oncolytic herpes simplex virus (oHSV)-1 have been shown to confer selectivity to cancer cells, which has enabled translational studies in humans. However, only a subset of cancer patients inoculated with oHSV-1 have shown objective response in phase 1 and 2 clinical trials. We have recently identified GBM lines that are resistant to oHSV mediated oncolysis. Based on our findings, we have created recombinant oHSV-TRAIL, an oHSV bearing a secretable pro-apoptotic tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), and hypothesize that oHSV-TRAIL could be used as a cancer therapeutic to target a broad spectrum of resistant tumors in a mechanism based manner. In this proposal, we will screen a panel of glioma stem cells (GSC) isolated from patient derived primary GBM lines, for their sensitivity to oHSV and TRAIL and identify lines that are resistant to oHSV or both oHSV and TRAIL. Based on our preliminary studies on the downregulation of ERK, activation of JNK and p38 and subsequent up- regulation of caspases in oHSV and TRAIL resistant glioma cells treated with oHSV-TRAIL, we hypothesize that oHSV will target cell proliferation pathway and prime oHSV and/or TRAIL resistant established glioma cells and primary GSCs to TRAIL mediated apoptosis. The efficacy of oHSV-S-TRAIL will be evaluated in vitro and in mouse models created from identified lines and ultimately in resistant lines identified from screening a panel of freshly resected patient tumors. The incorporation of genetically engineered fluorescent (GFP and mCherry) and bioluminescent (firefly luciferase; Fluc and Renilla luciferase;Rluc) imaging markers into oHSV and glioma cells will allow us to follow delivery and spread of oHSV in vivo; changes in glioma volumes and invasion; and the efficacy of oHSV-TRAIL by in vivo bioluminescence imaging (BLI) and intravital microscopy (IVM). We anticipate that our findings will have a major contribution towards: 1) elucidating molecular mechanisms of how oHSV mediated oncolysis and TRAIL can function in concert to target heterogeneous glioma cells which are resistant to viral oncolysis and/or cytotoxic therapies; and 2) ultimately developing novel oHSV based therapies for patients with brain tumors.