Studies in Poxvirus Host Range Genes and Tropism

痘病毒宿主范围基因和趋向性研究

基本信息

批准号：
7786263
负责人：
Grant McFadden
金额：
$ 35.87万
依托单位：
UNIVERSITY OF FLORIDA
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-03-16 至 2014-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7786263
关键词：
Animal Model Australia Benign Biological Models Cancer Control Cells Chemotherapy-Oncologic Procedure Collection Complex Cullin 1 Development Disease Drug usage Embryo European Fibroblasts Gene Targeting Genes Glioma Human Immunocompetent Immunodeficient Mouse Infection Interferons Knock-out Lagomorpha Libraries Malignant Neoplasms Mediating Molecular Mus Mutagenesis Myxoma virus New Zealand Oncolytic Oryctolagus cuniculus Pathogenesis Pathway interactions Pharmaceutical Preparations Phosphotransferases Pike fish Population Poxviridae Proliferating Protein Microchips Proteins Reporting Role Signal Pathway Signal Transduction Signaling Molecule Sirolimus Small Interfering RNA Species Specificity T-Lymphocyte Testing Therapeutic Tropism Tumor Necrosis Factor-alpha Tumor Necrosis Factors Viral Viral Proteins Virus Virus Diseases Virus Replication Work Xenograft procedure analog cancer cell cancer type enzootic feral inhibitor/antagonist killings knockout gene man member novel oncolysis permissiveness pre-clinical protein protein interaction public health relevance response skin lesion tool tumor virus tropism

项目摘要

DESCRIPTION (provided by applicant): We propose to examine the molecular pathways that mediate the species specificity and tropism of one particular poxvirus, namely myxoma virus (MV). MV is a rabbit-specific poxvirus that induces distinctly different disease profiles depending on the specific rabbit species but is nonpathogenic for every other host species, including man. Our lab has extensively studied MV and the disease it causes in rabbits, called myxomatosis, as a model system to investigate the fundamental principles of poxvirus pathogenesis, particularly by exploiting our large and growing collection of targeted MV gene knockout constructs. Several years ago, we unexpectedly discovered that primary mouse cells, which are normally nonpermissive for MV infection, could be rendered fully permissive by interrupting the cellular interferon (IFN) responses. This work then led to the discovery that the majority of human cancer cells tested were fully permissive for MV and that MV is remarkably effective as oncolytic therapy for the treatment of human cancer xenografts in a variety of animal models. We also discovered that two viral host range factors (M-T5 and M063) were also critical for permissive MV replication in many human cancer cells and we could manipulate viral permissiveness and oncolysis through these viral proteins and their host cell protein targets. Finally, our most recent observation related to this proposal is that primary human cells are protected from MV infection synergistically by IFN and tumor necrosis factor (TNF). We propose to: 1- Evaluate the roles of viral host range factors in MV tropism for human cancer cells. We propose to analyze the interactions of MV host range proteins with host cell signaling molecules (like Akt) by mutagenesis, siRNA knockdowns, signaling inhibitors and protein microarrays. We will test a number of adjunct strategies, like signaling modifier drugs used for cancer chemotherapy in man, to increase MV oncolytic potential for a wider spectrum of human cancer cells. 2- Investigate the role of host IFN and TNF responses in MV tropism. We will study MV knockout viruses that are deleted in several key viral host range genes to investigate their roles in inhibiting IFN or TNF responses, and modulating MV tropism and oncolysis. This new information will allow for more rational approaches to optimizing MV virotherapy against a wider spectrum of human cancers, and for controlling MV replication in primary noncancerous human cells. PUBLIC HEALTH RELEVANCE: Previously, our studies on poxvirus pathogenesis were more focused on studying the basic mechanisms by which one particular rabbit-specific poxvirus called myxoma virus (MV) causes disease in the rabbit host. Recently, we discovered that MV also infects and kills a wide spectrum of human cancer cells and we have now used MV to successfully treat several types of cancers in animal models. In this proposal, we will study two key viral host range factors and their cellular signaling pathway targets in order to assist the preclinical development of MV as a new oncolytic therapeutic for cancer in man.

描述（由申请人提供）：我们建议检查介导一种特定的一种特定蛇毒的物种特异性和向量的分子途径，即粘液瘤病毒（MV）。 MV是一种兔子特异性的痘病毒，根据特定的兔子物种诱导明显不同的疾病特征，但对于包括人在内的所有其他宿主物种而言都是非致病性的。我们的实验室已经广泛研究了MV及其在兔子中引起的疾病，称为粘膜瘤病，是一种模型系统，以研究痘病毒发病机理的基本原理，尤其是通过利用我们大量且越来越多的目标MV基因敲除结构的收集。几年前，我们出乎意料地发现，通常不接受MV感染的原代小鼠细胞可以通过中断细胞干扰素（IFN）反应来完全允许。然后，这项工作导致发现，大多数被测试的人类癌细胞具有完全允许的MV，并且MV非常有效，因为在多种动物模型中治疗人类癌症异种移植物治疗人溶治疗。我们还发现，两个病毒宿主范围因子（M-T5和M063）对于许多人类癌细胞中的允许MV复制也至关重要，我们可以通过这些病毒蛋白及其宿主细胞蛋白靶标操纵病毒允许和撞击。最后，我们与该提案有关的最新观察结果是，原代人细胞受到IFN和肿瘤坏死因子（TNF）协同性的MV感染。我们建议：1-评估病毒宿主范围因子在MV向热带中的人类癌细胞中的作用。我们建议通过诱变，siRNA敲低，信号抑制剂和蛋白质微阵列分析MV宿主范围蛋白与宿主细胞信号分子（如AKT）的相互作用。我们将测试许多辅助策略，例如用于人类的癌症化学疗法的信号修饰药，以提高MV溶瘤潜力，以增加更多的人类癌细胞。 2-调查宿主IFN和TNF响应在MV tropism中的作用。我们将研究在几个关键病毒宿主范围基因中删除的MV基因敲除病毒，以研究其在抑制IFN或TNF反应中的作用，并调节MV的Tropism和Oncolysiss。这些新信息将允许采用更合理的方法来优化针对更广泛的人类癌症的MV病毒疗法，并控制原代非癌性人类细胞中的MV复制。公共卫生相关性：以前，我们对痘病毒发病机理的研究更加专注于研究一种基本机制，通过这种机制，一种特定的兔特异性蛇毒称为粘液瘤病毒（MV）在兔宿主中引起疾病。最近，我们发现MV还感染并杀死了广泛的人类癌细胞，现在我们已经使用MV成功地治疗了动物模型中的几种类型的癌症。在此提案中，我们将研究两个关键的病毒宿主范围及其细胞信号通路靶标，以帮助MV作为人类癌症的新型溶瘤治疗。