Myxoma Virus (MV) Oncolysis for treating human cancer

粘液瘤病毒 (MV) 溶瘤治疗人类癌症

• 批准号: 8413599
• 负责人: Grant McFadden
• 金额: $ 27.72万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-03-01 至 2014-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8413599
• 关键词: Acute Myelocytic Leukemia; Animal Model; Antibodies; Autologous Bone Marrow Transplantation; Autologous Stem Cell Transplantation; B-Cell Lymphomas; Blood; Bone Marrow; CSF3 gene; Cancer Model; Cancer Patient; Cancer cell line; Cells; Clinic; Clinical Trials; Data; Development; Engraftment; Florida; Hematopoietic; Hematopoietic stem cells; High Dose Chemotherapy; Human; Image; Immune; Immune system; Immunocompetent; Immunodeficient Mouse; In Vitro; Infection; Injection of therapeutic agent; Leukocytes; Libraries; Luciferases; Malignant Neoplasms; Malignant neoplasm of brain; Metastatic Melanoma; Molecular; Multipotent Stem Cells; Mus; Myxoma virus; Normal tissue morphology; Oncolytic; Oryctolagus cuniculus; Patients; Peripheral Blood Mononuclear Cell; Peripheral Blood Stem Cell; Pharmaceutical Preparations; Phosphoproteins; Poxviridae; Proteins; Protocols documentation; Recombinants; Refractory; Reporter; Safety; Sampling; Signal Transduction; Sirolimus; Source; Stem cells; Testing; Therapeutic; Tissues; Transplantation; Tropism; Virus; Virus Diseases; Virus Receptors; Xenograft procedure; base; cancer cell; cancer type; cytokine; human stem cells; in vitro Assay; in vivo; killings; leukemia; leukemia/lymphoma; man; mutant; neoplastic cell; next generation; novel therapeutics; oncolysis; pre-clinical; prevent; purge; recombinant virus; reconstitution; response; stem; stem cell differentiation; therapeutic transgene; tumor; tumorigenic; virus genetics; virus tropism

Project Summary Myxoma virus (MV) infects only rabbits in vivo, but also has a natural capacity to infect a wide variety of human cancer cells in vitro and in vivo. Thus, MV is an attractive candidate for oncolytic virotherapy to treat human cancer. MV has been used to successfully treat several diverse human brain cancers in xenografted immunodeficient mice and murine metastatic melanoma in immunocompetent mice. Here, MV will be developed for human clinical trials by exploiting a therapeutic strategy for which the virus is uniquely well- suited: ex vivo purging of cancer cells within human bone marrow or mobilized-PBMC samples from patients who would normally be excluded from autologous stem cell transplantation following high dose chemotherapy. Considerable preliminary data has been collected to support this proposal: 1) MV does not perturb or compromise human multipotent stem cell differentiation in immunodeficient mice engrafted with normal human bone marrow or cytokine-mobilized PBMCs, 2) MV eliminates a wide variety of human cancer cells following ex vivo purging, 3) MV can effectively purge not only permissive leukemia/lymphoma cells in vivo, but also unexpectedly prevents even nonpermissive human leukemia cells (such as KG1 cells) from engraftment or tumor induction, and 4) MV recombinants that express a variety of useful reporter proteins (fluorescent and bioluminescent) for imaging purposes have already been constructed. Specifically, our aims are: 1) Validate MV safety for ex vivo treatment of normal human hematopoietic stem cells: The safety of MV-purging for normal human stem cell differentiation will be tested, using engrafted immunodeficient NOG mice to verify full hematopoietic cell engraftment and immune reconstitution. MV purging will be tested on primary human stem/progenitor cells derived from normal bone marrow and G-CSF-mobilized PBMCs, using hematopoietic colony forming cell assays in vitro as well as for efficient hematologic cell engraftment in vivo. 2) Optimize MV ex vivo cancer cell purging: Two human cancers, B-cell lymphoma and acute myeloid leukemia, will be investigated for the ability of ex vivo MV purging to eliminate their tumorigenic potential in vivo in engrafted NOG mice. The cancer cells and viruses will be tagged with distinguishable luciferases that allow the engrafted tumor cells and the therapeutic virus to be independently tracked in vivo. Primary cells from acute myeloid leukemia patients will also be tested for the ability of MV to specifically eliminate the contaminating cancer cells and allow the selective engraftment of only noncancerous human leukocytes. 3) Investigate the mechanism of MV purging of primary human leukemia cells: We have recently shown that ex vivo infection of human KG1 leukemia cells with MV prevents the subsequent engraftment and tumor formation of these cells into NOG recipient mice, despite the fact that these cells are completely nonpermissive for MV infection in vitro. To assess for virus-induced cell signaling changes, we probed MV-infected KG1 cells with an array of antibodies to 46 different human signaling phosphoproteins, and observed that MV infection specifically induces Stat5 and Hck activation in KG1 cells. We will explore the functional significance of these host cell signaling activations for the successful ex vivo tumor cell purging of human leukemia calls by MV.

项目概要 粘液瘤病毒（MV）仅在体内感染兔子，但也具有感染多种人类的天然能力 体外和体内的癌细胞。因此，MV 是溶瘤病毒疗法治疗人类的一个有吸引力的候选者。 癌症。 MV 已成功用于治疗多种异种移植的人类脑癌 免疫缺陷小鼠和免疫功能正常小鼠的鼠转移性黑色素瘤。这里，MV将是 通过利用该病毒独特的治疗策略，为人类临床试验而开发 适用：离体清除人骨髓内的癌细胞或患者的动员 PBMC 样本 通常在高剂量化疗后被排除在自体干细胞移植之外的人。 已经收集了大量的初步数据来支持这一提议：1）MV 不会扰动或 损害移植正常人类免疫缺陷小鼠的人类多能干细胞分化 骨髓或细胞因子动员的 PBMC，2) MV 消除多种人类癌细胞 体内清除，3）MV不仅可以有效清除体内允许的白血病/淋巴瘤细胞，还可以 出乎意料地阻止甚至不允许的人类白血病细胞（例如 KG1 细胞）的植入或 肿瘤诱导，以及 4) 表达多种有用报告蛋白（荧光和荧光蛋白）的 MV 重组体 用于成像目的的生物发光）已经被构建。具体来说，我们的目标是： 1) 验证正常人造血干细胞离体治疗的MV安全性： 将使用移植的免疫缺陷 NOG 测试正常人类干细胞分化的 MV 清除 小鼠验证完整的造血细胞植入和免疫重建。 MV 吹扫将在 源自正常骨髓和 G-CSF 动员的 PBMC 的原代人类干/祖细胞，使用 体外造血集落形成细胞测定以及体内有效的血细胞植入。 2) 优化 MV 离体癌细胞清除：两种人类癌症，B 细胞淋巴瘤和急性骨髓瘤 白血病，将研究离体 MV 清除以消除体内致瘤潜力的能力 在移植的 NOG 小鼠中。癌细胞和病毒将被标记上可区分的荧光素酶，从而允许 移植的肿瘤细胞和治疗病毒在体内进行独立追踪。原代细胞来自 急性髓系白血病患者还将接受MV特异性消除白血病的能力测试。 污染癌细胞并仅允许选择性植入非癌性人类白细胞。 3）研究原代人白血病细胞MV清除的机制：我们最近证明 用 MV 离体感染人 KG1 白血病细胞可防止随后的植入和肿瘤 这些细胞形成 NOG 受体小鼠，尽管事实上这些细胞是完全不允许的 用于体外MV感染。为了评估病毒诱导的细胞信号传导变化，我们探测了 MV 感染的 KG1 细胞 使用针对 46 种不同人类信号磷蛋白的一系列抗体，并观察到 ​​MV 感染 特异性诱导 KG1 细胞中 Stat5 和 Hck 激活。我们将探讨这些的功能意义 MV 成功体外清除人类白血病细胞的宿主细胞信号传导激活。