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.

编号57-“黑色素瘤的细小病毒疗法” Peter Tattersall，博士(实验室医学)，Ruth Halaban，博士(皮肤科) 资格类别：在皮肤病方面没有工作经验的资深研究人员 我们已经从五种细小病毒的筛选中确定了孤儿细小病毒Lulll为有效的溶瘤病毒 在细胞培养中感染逐步转化的黑素细胞的物种。我们已经证明了Lulll 有效地感染这些细胞，表达丰富的病毒结构蛋白和非结构蛋白。此外， Lulll产生大量的子代病毒粒子，这些病毒粒子迅速传播到并杀死其他转化的细胞。 文化。这项研究的进一步目标是产生和表征复制能力 Lulll的衍生物作为病毒治疗剂治疗恶性黑色素瘤的潜力。我们将寻求 通过检测Lulll在人黑素细胞系中的高效生长能力来确认其适用性 通过不同的循序渐进的程序转变。Lulll也将被评估其有效感染的能力 正常黑素细胞与一组原代和已建立的黑色素瘤细胞培养的比较，以探索其 治疗指标及其嗜黑作用的通用性。我们会进一步加强市民大众的选择 通过基因操作在肿瘤转化的人类黑素细胞培养中的Lulll衍生物。为了这个， 我们将构建Lulll P4启动子变体，预测其对Notch信号的激活有反应 途径，黑色素瘤进化过程中的一种常见事件。同时，我们将使用Lulll P4启动子衍生物 为了产生取代Ets1位点的简并寡核苷酸序列的具有感染性的克隆库， 转化成纤维细胞肿瘤选择性的主要决定因素。此库将用于尝试选择 带有P4近端转录位点的突变体(S)进一步优化了在完全转化的黑素细胞中的生长。 将对选定的突变体进行筛选，以了解它们在黑色素瘤肿瘤细胞中的扩张和杀伤能力，而 避免那些来自正常的原代黑素细胞培养的平行小组。 病毒通常对它们将感染和杀死的宿主细胞类型具有很强的特异性。这个项目将探索是否 Lulll是细小病毒家族中一种相对黑色素瘤特异性的成员，可以进一步修饰，以便 优先摧毁恶性黑色素瘤细胞，而不是正常细胞。