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。没有治愈方法，目前的治疗方法只能延长几个月的生命，往往以牺牲生活质量为代价。据报道，许多细小病毒显示出对癌细胞的溶瘤潜力，大鼠细小病毒H-1目前正在临床试验中治疗神经胶质瘤患者。在我们最初对大量细小病毒进行筛查的工作中，我们发现了一种相对不起眼的细小病毒LuIII，它的表现比包括H-1在内的任何其他测试病毒都要好得多，并且是唯一一种成功杀死多种人类神经胶质瘤的细小病毒。LuIII在大脑中似乎是安全的，并且不靶向或杀死神经元，并且显示出对正常胶质细胞的最小感染。在这里，我们测试了一些假设有关的能力LuIII选择性感染，复制，杀死人类胶质瘤。 在第一组实验中，我们测试了LuIII将成功靶向并杀死移植到脑中的胶质母细胞瘤细胞的假设，在瘤内病毒注射后和静脉内接种后，对正常脑的毒性相对较小。通过表达红色荧光报告基因检测肿瘤，通过对LuIII病毒抗原NS 1进行绿色免疫荧光染色检测病毒感染的细胞。神经胶质瘤的一个关键的临床问题是手术或放射治疗后复发，在这里，我们测试的假设，LuIII可能会保持一个无症状的低水平的感染在正常的人类神经胶质细胞，LuIII所产生的正常细胞，从而减弱或阻止神经胶质瘤细胞的扩张或复发。我们将证实我们的初步研究结果和测试的假设，LuIII不产生抗病毒干扰素（IFN）的反应，LuIII感染是NT减弱IFN。这种与IFN系统的独立性使LuIII与脑中使用的许多其他对IFN敏感的溶瘤病毒不同。LuIII对IFN的不敏感性将允许LuIII和IFN的共同治疗（IFN是一种部分有效的抗肿瘤治疗， 脑肿瘤的子集），或用LuIII与IFN-敏感性溶瘤病毒共同治疗。我们将采用深度全外显子组基因测序和mRNA转录组测序，以寻找与高水平LuIII感染相关的胶质瘤基因突变。这种遗传信息对于理解LuIII选择性感染神经胶质瘤的机制是有用的，并且也可以证明其作为肿瘤相关突变最可能与高LuIII感染相关的诊断预测因子是有用的。最后，我们的初步数据表明，LuIII不仅能有效靶向和杀死胶质母细胞瘤，还能感染和杀死其他侵入大脑的癌症，包括黑色素瘤和肺癌。肺癌转移是最常见的问题，涉及脑中的继发性癌症。我们将在与上述涉及神经胶质瘤的实验平行的实验中测试LuIII靶向脑中的肺癌细胞的假设。我们的中心目标是测试LuIII作为基本上减弱或破坏人类脑肿瘤的安全有效手段的潜力。

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.