Parvoviral Virothereapy for Melanoma

黑色素瘤的细小病毒病毒治疗

• 批准号: 7608590
• 负责人: Peter J. Tattersall
• 金额: $ 5.32万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-04-01 至 2009-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7608590
• 关键词: Categories; Cells; Cultured Cells; Dermatology; Doctor of Philosophy; Eligibility Determination; Event; Evolution; Family; Fibroblasts; Genetic Transcription; Growth; Human; Laboratories; Libraries; Medicine; Melanoma Cell; Notch Signaling Pathway; Oligonucleotides; Oncolytic viruses; Orphan; Parvovirus; Procedures; Research; Research Personnel; Site; Structural Protein; Therapeutic Index; Transformed Cell Line; Variant; Viral; Virion; Virus; Work; cell transformation; cell type; genetic manipulation; killings; melanocyte; melanoma; member; mutant; neoplastic cell; promoter; skin disorder

P/F #57 - "Parvoviral Virotherapy for Melanoma" Peter Tattersall, PhD (Laboratory Medicine), Ruth Halaban, PhD (Dermatology) Eligibility Category: Established investigator with no previous work in skin diseases We have identified the orphan parvovirus Lulll as an effective oncolytic virus from a screen of five parvoviral species infecting step-wise transformed melanocytes in cell culture. We have demonstrated that Lulll efficiently infects these cells, expressing abundant viral structural and non-structural proteins. Furthermore, Lulll generates significant amounts of progeny virions that rapidly spread to, and kill, other transformed cells in the culture. The further aims of this research are to generate and characterize replication competent derivatives of Lulll for their potential as virotherapeutic agents against malignant melanoma. We will seek to confirm the applicability of Lulll by examining its ability to grow productively in human melanocyte cell lines transformed by different stepwise procedures. Lulll will also be assessed for its ability to productively infect normal melanocytes versus a panel of primary and established melanoma cell cultures, in order to explore its therapeutic index and the generality of its melanotropism. We will further enhance the oncoselectivity of the Lulll derivatives in neoplastically transformed human melanocyte cultures by genetic manipulation. For this, we will construct Lulll P4 promoter variants predicted to be responsive to activation of the Notch signaling pathway, a frequent event in the evolution of melanoma. In parallel, we will use a Lulll P4 promoter derivative to generate an infectious clone library of degenerate oligonucleotide sequences replacing the Ets1 site, a major determinant of oncoselectivity in transformed fibroblasts. This library will be used to attempt selection of mutants with P4 proximal transcription site(s) further optimized for growth in fully transformed melanocytes. Selected mutants will be screened for their ability to expand in and kill a panel of melanoma tumor cells, while sparing those from a parallel panel of normal, primary melanocyte cultures. Viruses are often very specific for the host cell types they will infect and kill. This project will explore whether Lulll, a relatively melanoma-specific member of the parvovirus family, can be further modified so as to preferentially destroy malignant melanoma cells, compared to their normal counterparts.

P/F #57 -“细小病毒病毒治疗黑色素瘤” Peter Tattersall博士（实验室医学），Ruth Halaban博士（皮肤科） 资格类别：已确立的研究者，以前没有皮肤病方面的工作 我们已经从五种细小病毒的筛选中鉴定出孤儿细小病毒Lulll作为有效的溶瘤病毒。 种感染细胞培养物中逐步转化的黑素细胞。我们已经证明， 有效地感染这些细胞，表达丰富的病毒结构和非结构蛋白。此外，委员会认为， Lulll产生大量的子代病毒体，这些病毒体迅速扩散到并杀死其他转化细胞， 文化。本研究的进一步目的是产生和表征复制能力 Lulll的衍生物作为抗恶性黑素瘤的病毒治疗剂的潜力。我们将寻求 通过检测Lulll在人类黑素细胞系中高效生长的能力来证实其适用性 通过不同的逐步程序转化。还将评估Lulll的生产性感染能力 正常黑素细胞与一组原代和已建立的黑色素瘤细胞培养物，以探索其 治疗指数及其促黑作用的一般性。我们将进一步增强肿瘤的选择性， 通过遗传操作在肿瘤转化的人类黑素细胞培养物中的Lulll衍生物。为此， 我们将构建预测响应于Notch信号传导激活的LullI P4启动子变体 这是黑色素瘤发展过程中的一个常见事件。同时，我们将使用Lulll P4启动子衍生物 为了产生置换Ets 1位点的简并寡核苷酸序列的感染性克隆文库， 转化成纤维细胞中肿瘤选择性的主要决定因素。此库将用于尝试选择 具有P4近端转录位点的突变体进一步优化以在完全转化的黑素细胞中生长。 将筛选选定的突变体，以确定它们在一组黑色素瘤肿瘤细胞中扩增并杀死它们的能力， 将其与正常的原代黑素细胞培养物的平行组分开。 病毒通常对它们将感染和杀死的宿主细胞类型非常特异。该项目将探讨是否 Lulll是细小病毒家族的相对黑素瘤特异性的成员，可以进一步修饰以 与正常的黑色素瘤细胞相比，它们优先破坏恶性黑色素瘤细胞。