Jaime-Ramirez Alena Cristina, Dmitrieva Nina, Yoo Ji Young, Banasavadi-Siddegowda Yeshavanth, Zhang Jianying, Relation Theresa, Bolyard Chelsea, Wojton Jeffrey, Kaur Balveen
Department of Neurological Surgery, The Ohio State University Comprehensive Cancer Center, The James Cancer Hospital, and Solove Research Institute, Columbus, OH, USA.
Center for Biostatistics Biomedical Informatics Department, The Ohio State University Comprehensive Cancer Center, The James Cancer Hospital, and Solove Research Institute, Columbus, OH, USA.
J Gene Med. 2017 Mar;19(3). doi: 10.1002/jgm.2942.
Malignant gliomas (glioblastomas; GBMs) are extremely aggressive and have a median survival of approximately 15 months. Current treatment modalities, which include surgical resection, radiation and chemotherapy, have done little to prolong the lives of GBM patients. Chondroitin sulfate proteoglycans (CSPG) are critical for cell-cell and cell-extracellular matrix (ECM) interactions and are implicated in glioma growth and invasion. Chondroitinase (Chase) ABC is a bacterial enzyme that cleaves chondroitin sulfate disaccharide chains from CSPGs in the tumor ECM. Wild-type Chase ABC has limited stability and/or activity in mammalian cells; therefore, we created a mutant humanized version (Chase M) with enhanced function in mammalian cells.
We hypothesized that disruption of cell-cell and cell-ECM interactions by ChaseM and temozolomide (TMZ) will enhance the chemotherapeutic availability and sensitivity of glioma cells.
Utilizing primary patient-derived neurospheres, we found that ChaseM decreases glioma neurosphere aggregation in vitro. Furthermore, an oncolytic HSV-1 virus expressing secreted ChaseM (OV-ChaseM) enhanced viral spread and glioma cell killing compared to OV-Control, in vitro. OV-ChaseM plus TMZ combinatorial treatment resulted in a significant synergistic enhancement of glioma cell killing accompanied by an increase in apoptotic cell death. Intracellular flow cytometric analysis revealed a significant reduction in the phosphorylation of the pro-survival AKT protein following OV-ChaseM plus TMZ treatment. Lastly, in nude mice bearing intracranial GBM30 glioma xenografts, intratumoral OV-ChaseM plus TMZ (10 mg/kg by oral gavage) combination therapy resulted in a significant (p < 0.02) enhancement of survival compared to each individual treatment alone.
These data reveal that OV-ChaseM enhances glioma cell viral susceptibility and sensitivity to TMZ.
恶性胶质瘤（胶质母细胞瘤；GBM）极具侵袭性，中位生存期约为15个月。目前的治疗方式，包括手术切除、放疗和化疗，在延长GBM患者生命方面收效甚微。硫酸软骨素蛋白聚糖（CSPG）对细胞间和细胞与细胞外基质（ECM）的相互作用至关重要，并与胶质瘤的生长和侵袭有关。软骨素酶（Chase）ABC是一种细菌酶，可从肿瘤ECM中的CSPG上切割硫酸软骨素二糖链。野生型Chase ABC在哺乳动物细胞中的稳定性和/或活性有限；因此，我们创建了一个在哺乳动物细胞中功能增强的突变体人源化版本（Chase M）。
我们假设ChaseM和替莫唑胺（TMZ）破坏细胞间和细胞与ECM的相互作用将提高胶质瘤细胞的化疗可用性和敏感性。
利用原发性患者来源的神经球，我们发现ChaseM在体外可减少胶质瘤神经球聚集。此外，与OV-Control相比，表达分泌型ChaseM的溶瘤性单纯疱疹病毒1型（OV-ChaseM）在体外增强了病毒传播和胶质瘤细胞杀伤。OV-ChaseM加TMZ联合治疗导致胶质瘤细胞杀伤显著协同增强，同时凋亡细胞死亡增加。细胞内流式细胞术分析显示，OV-ChaseM加TMZ治疗后，促生存AKT蛋白的磷酸化显著降低。最后，在携带颅内GBM30胶质瘤异种移植物的裸鼠中，瘤内注射OV-ChaseM加TMZ（通过口服灌胃给予10mg/kg）联合治疗与单独的每种治疗相比，显著（p<0.02）提高了生存率。
这些数据表明，OV-ChaseM增强了胶质瘤细胞对病毒的易感性和对TMZ的敏感性。
