Lal Sangeet, Raffel Corey
Department of Neurological Surgery, University of California San Francisco, San Francisco, CA, USA.
Methods Mol Biol. 2022;2423:165-177. doi: 10.1007/978-1-0716-1952-0_16.
Oncolytic virotherapy translational research in the current era is heavily focused on the interaction of the immune system and tumor microenvironment with oncolytic viruses. Preclinical xenograft studies using human cells in immunodeficient mouse models does not serve this purpose. As a consequence, developing syngeneic immunocompetent murine cancer models sensitive to infection and growth of specific oncolytic viruses is required. The group 3 subtype of medulloblastoma, among the four molecular subgroups-WNT, SHH, Group 3, and Group 4, has the worst prognosis and the poorest outcome. Sadly, current treatments cause long-term toxicity and morbidity to survivors adversely affecting their quality of life. Alternate effective therapy with less side effects is urgently needed. We have shown that oncolytic measles virus (MV) is effective against localized as well as CSF-disseminated medulloblastoma in immunodeficient mouse models. To study the interaction of immune system with oncolytic measles virotherapy, we have developed a murine group 3 medulloblastoma cell line (CSCG) that is infectible by MV, is killed by MV, allows replication of MV, and is tumorigenic in the brain of syngeneic transgenic immune-competent mice. Intratumoral injection of MV results in significant prolongation of survival in mice bearing CSCG tumors in the brain. This model provides the first suitable platform to examine therapeutic regimens of MV therapy for MB tumors in the presence of intact immune system. Here, we describe our lab protocols to develop this cell line and the mouse model.
当前时代的溶瘤病毒疗法转化研究主要聚焦于免疫系统和肿瘤微环境与溶瘤病毒的相互作用。在免疫缺陷小鼠模型中使用人类细胞进行的临床前异种移植研究无法达到这一目的。因此，需要开发对特定溶瘤病毒的感染和生长敏感的同基因免疫活性小鼠癌症模型。在髓母细胞瘤的四个分子亚组——WNT、SHH、3组和4组中，3组亚型的预后最差，结局也最糟糕。遗憾的是，目前的治疗方法会给幸存者带来长期毒性和发病率，对他们的生活质量产生不利影响。迫切需要副作用较小的替代有效疗法。我们已经表明，溶瘤麻疹病毒（MV）在免疫缺陷小鼠模型中对局部以及脑脊液播散的髓母细胞瘤有效。为了研究免疫系统与溶瘤麻疹病毒疗法的相互作用，我们开发了一种3组髓母细胞瘤小鼠细胞系（CSCG），它可被MV感染、能被MV杀死、允许MV复制，并且在同基因转基因免疫活性小鼠的大脑中具有致瘤性。瘤内注射MV可显著延长大脑中携带CSCG肿瘤的小鼠的生存期。该模型为在完整免疫系统存在的情况下研究MV治疗MB肿瘤的治疗方案提供了首个合适的平台。在此，我们描述我们开发这种细胞系和小鼠模型的实验室方案。
