LuIII parvovirus targets glioma

描述(申请人提供)：胶质母细胞瘤是侵袭性和侵袭性的脑肿瘤，通常会导致一年内死亡。目前还没有治愈方法，目前的治疗方法只能延长生命几个月，通常是以牺牲生活质量为代价的。据报道，一些细小病毒对癌细胞具有溶瘤潜力，而一种名为H-1的大鼠细小病毒目前正在进行临床试验，用于治疗胶质瘤患者。在我们最初的工作中，我们筛选了大量的细小病毒，我们发现了一种相对鲜为人知的细小病毒LuIII，它的表现比包括H-1在内的任何其他测试的病毒都要好得多，也是唯一一种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，不会靶向或杀死神经元，对正常胶质细胞的感染很小。在这里，我们测试了一些与LuIII选择性感染、复制和杀死人类胶质瘤的能力相关的假设。在第一组实验中，我们测试了这样的假设，即LuIII将成功地靶向并杀死移植到大脑中的胶质母细胞瘤细胞，对正常大脑的毒性相对较小，无论是在瘤内注射病毒之后，还是在静脉接种之后。肿瘤是通过表达红色荧光报告来检测的，病毒感染的细胞是通过LuIII病毒抗原NS1的绿色免疫荧光染色来检测的。胶质瘤的一个关键临床问题是手术或放射治疗后的复发；在这里，我们假设LuIII可能在正常的人神经胶质细胞中保持无症状的低水平感染，而来自正常细胞的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，并测试LuIII不产生抗病毒干扰素(干扰素)反应的假设，以及LuIII感染被干扰素减弱的假设。这种对干扰素系统的独立性使LuIII有别于大脑中使用的其他一些对干扰素敏感的溶瘤病毒。LuIII对干扰素的不敏感将允许与LuIII和干扰素联合治疗(干扰素是一种部分有效的抗肿瘤治疗方法 脑肿瘤的一种亚型)，或与LuIII和对干扰素敏感的溶瘤病毒共同治疗。我们将使用深层完整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。这些遗传信息既有助于理解LuIII选择性感染胶质瘤的机制，也可能被证明是有用的诊断预测指标，即哪些肿瘤相关突变最有可能与LuIII高感染相关。最后，我们的初步数据表明，LuIII不仅在靶向和杀死胶质母细胞瘤方面有效，而且还可以感染和杀死其他侵袭大脑的癌症，包括黑色素瘤和肺癌。肺癌转移是脑部继发性癌症最常见的问题。我们将通过与上面涉及胶质瘤的实验平行的实验来检验LuIII以大脑中的肺癌细胞为靶点的假设。我们的中心目标是测试LuIII作为一种安全有效的手段大幅减弱或摧毁人类脑瘤的潜力。

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.