LuIII parvovirus targets glioma

描述（由申请人提供）：胶质母细胞瘤是侵袭性和侵袭性脑肿瘤，通常在诊断后一年内导致死亡。目前还没有治愈方法，目前的治疗方法只能延长几个月的生命，而且往往是以牺牲生活质量为代价的。据报道，许多细小病毒显示出对癌细胞的溶瘤潜力，一种大鼠细小病毒H-1目前正在临床试验中用于治疗胶质瘤患者。在我们最初筛选大量细小病毒的工作中，我们发现了一种相对模糊的细小病毒LuIII，它的表现明显优于其他任何测试，包括H-1，并且是唯一成功杀死多个人类胶质瘤的细小病毒。LuIII在大脑中似乎是安全的，不靶向或杀死神经元，对正常神经胶质的感染最小。在这里，我们测试了一些关于LuIII选择性感染、复制和杀死人类胶质瘤的能力的假设。在第一组实验中，我们验证了LuIII将成功靶向并杀死移植到大脑中的胶质母细胞瘤细胞的假设，在肿瘤内注射病毒和静脉注射后，对正常大脑的毒性相对较小。通过表达红色荧光报告蛋白检测肿瘤，通过LuIII病毒抗原NS1的绿色免疫荧光染色检测病毒感染细胞。胶质瘤的一个关键临床问题是手术或放疗后的复发；在这里，我们验证了这样一种假设，即LuIII可能在正常的人类神经胶质细胞中维持无症状的低水平感染，并且来自正常细胞的LuIII将因此减弱或阻止胶质瘤细胞的扩张或复发。我们将证实我们的初步发现，并验证LuIII不会产生抗病毒干扰素（IFN）反应的假设，以及IFN不会减弱LuIII感染的假设。这种对IFN系统的独立性使LuIII与大脑中对IFN敏感的许多其他溶瘤病毒区别开来。由于LuIII对IFN不敏感，因此可以与LuIII和IFN联合治疗（IFN是一种部分有效的抗肿瘤药物）

DESCRIPTION (provided by applicant): Glioblastomas are aggressive and invasive brain tumors that generally lead to death within a year of diagnosis. No cure is available, and current treatments prolong life by only a few months, often at the expense of quality of life. A number of parvoviruses have been reported to show oncolytic potential against cancer cells, and a rat parvovirus, H-1, is currently in clinical trials to treat glioma patients. In our initial work screning a large number of parvoviruses, we found one relatively obscure parvovirus, LuIII, that performed substantially better than any of the others tested, including H-1, and was the only parvovirus tested that successfully killed multiple human gliomas. LuIII appears safe in the brain, and does not target or kill neurons and shows minimal infection of normal glia. Here we test a number of hypotheses related to the ability of LuIII to selectively infect, replicate in, an kill human glioma. In the first set of experiments, we test the hypothesis that LuIII will successfully target and kill glioblastoma cells that are transplanted into the brain, with relativey little toxicity to the normal brain, both after an intratumoral virus injection, and after intravenus inoculation. Tumors are detected by expression of a red fluorescent reporter, and virally infected cells are detected by green immunofluorescent staining for the LuIII viral antigen NS1. A critical clinical problem with glioma is their recurrence after surgical or radiation treatment; here we tes the hypothesis that LuIII may maintain an asymptomatic low level of infection in normal human glial cells, and that LuIII arising from normal cells will thereby attenuate or block glioma cell expansion or recurrence. We will corroborate our preliminary findings and test the hypothesis that LuIII does not generate an anti-viral interferon (IFN) response, and that LuIII infection is nt attenuated by IFN. This independence from the IFN system sets LuIII apart from a number of other oncolytic viruses used in the brain which are sensitive to IFN. LuIII's insensitivity to IFN would allow a co-treatment with LuIII and IFN (IFN is a partially effective anti-tumor treatment in a subset of brain tumors), or co-treatment with LuIII together with an IFN-sensitive oncolytic virus. We will employ deep whole exome genetic sequencing, and sequencing of the mRNA transcriptome, to search for gene mutations in glioma that correlate with high levels of LuIII infection. This genetic information is useful both to understand the mechanisms underlying LuIII's selective infection of gliomas, and also may prove useful as a diagnostic predictor of which tumor-related mutations are most likely to be associated with a high LuIII infection. Finally, our preliminary data suggest that LuIII is effective at not only targeting and killing glioblastoma, but also infects and kills other cancers that invade the brain, including melanoma and lung cancer. Lung cancer metastasis is the most common problem involving secondary cancer in the brain. We will test the hypothesis that LuIII targets lung cancer cells in the brain n experiments parallel to those above involving glioma. Our central goal is to test the potential of LuIII as a safe and effective means of substantially attenuating or destroying brain tumors in humans.