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: 8036039
• 负责人: Grant McFadden
• 金额: $ 29.49万
• 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/8036039
• 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; public health relevance; purge; recombinant virus; reconstitution; response; stem; stem cell differentiation; therapeutic transgene; tumor; tumorigenic; virus genetics; virus tropism

DESCRIPTION (provided by applicant): 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. PUBLIC HEALTH RELEVANCE Recently, we discovered that one particular rabbit-specific poxvirus, called myxoma virus (MV), also infects and kills a wide spectrum of human cancer cells and MV has been used to successfully treat several types of cancers in animal models. In order to facilitate the preclinical development of MV as a new oncolytic therapeutic for cancer in man, we propose to exploit two specific cancer models (human B-cell lymphoma and acute myeloid leukemia) to establish the optimal conditions for a novel therapeutic "cancer cell purging" protocol. This strategy will benefit leukemia/lymphoma cancer patients who are currently excluded from autologous bone marrow transplantation by allowing their own stem cell samples (from bone marrow or blood) to be purged of cancer cells prior to re-engraftment and reconstitution of their immune system following high dose chemotherapy.

描述（由申请人提供）：粘液瘤病毒（MV）仅在体内感染兔子，但也具有在体外和体内感染多种人类癌细胞的天然能力。因此，MV 是治疗人类癌症的溶瘤病毒疗法的一个有吸引力的候选者。 MV 已成功用于治疗异种移植免疫缺陷小鼠中的多种人类脑癌和免疫功能正常小鼠中的鼠转移性黑色素瘤。在这里，MV 将通过利用该病毒独特适合的治疗策略来开发用于人类临床试验：离体清除人骨髓内的癌细胞或来自通常被排除在自体干细胞之外的患者的动员 PBMC 样本高剂量化疗后的细胞移植。已经收集了大量的初步数据来支持这一提议：1) MV 不会干扰或损害移植了正常人骨髓或细胞因子动员的 PBMC 的免疫缺陷小鼠的人类多能干细胞分化，2) MV 消除多种人类癌细胞离体清除后，3）MV不仅可以有效地清除体内允许的白血病/淋巴瘤细胞，而且还可以出乎意料地预防甚至不允许的人类细胞来自植入或肿瘤诱导的白血病细胞（例如 KG1 细胞），以及 4）表达各种用于成像目的的有用报告蛋白（荧光和生物发光）的 MV 重组体已经构建。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：将测试 MV 清除对正常人干细胞分化的安全性，使用移植的免疫缺陷 NOG 小鼠来验证完整的造血细胞移植和免疫重建。将使用体外造血集落形成细胞测定以及体内有效的血液细胞植入，在源自正常骨髓和 G-CSF 动员的 PBMC 的原代人干/祖细胞上测试 MV 清除。 2) 优化 MV 离体癌细胞清除：将研究两种人类癌症（B 细胞淋巴瘤和急性髓性白血病）离体 MV 清除在移植的 NOG 小鼠体内消除其致瘤潜力的能力。癌细胞和病毒将被标记上可区分的荧光素酶，从而可以在体内独立追踪移植的肿瘤细胞和治疗病毒。还将测试来自急性髓系白血病患者的原代细胞的 MV 特异性消除污染性癌细胞并允许选择性植入非癌性人类白细胞的能力。 3）研究MV清除原代人白血病细胞的机制：我们最近表明，用MV离体感染人KG1白血病细胞可阻止这些细胞随后植入NOG受体小鼠并形成肿瘤，尽管事实上这些细胞体外完全不允许 MV 感染。为了评估病毒诱导的细胞信号传导变化，我们用针对 46 种不同人类信号传导磷蛋白的一系列抗体探测了 MV 感染的 KG1 细胞，并观察到 ​​MV 感染特异性诱导 KG1 细胞中 Stat5 和 Hck 激活。我们将探讨这些宿主细胞信号传导激活对于 MV 成功离体肿瘤细胞清除人类白血病细胞的功能意义。 公共健康相关性 最近，我们发现一种特殊的兔子特异性痘病毒，称为粘液瘤病毒 (MV)，也能感染并杀死多种人类癌细胞，并且 MV 已被用于在动物模型中成功治疗多种类型的癌症。为了促进 MV 作为一种新的人类癌症溶瘤疗法的临床前开发，我们建议利用两种特定的癌症模型（人类 B 细胞淋巴瘤和急性髓性白血病）来建立新型治疗“癌细胞”的最佳条件。净化”协议。该策略将使目前被排除在自体骨髓移植之外的白血病/淋巴瘤癌症患者受益，方法是允许他们自己的干细胞样本（来自骨髓或血液）在重新植入和重建免疫系统之前清除癌细胞。高剂量化疗。