Glioma immunotherapy with strategies based on autonomous parvovirus MVMp

基于自主细小病毒 MVMp 的神经胶质瘤免疫治疗策略

• 批准号: 9056439
• 负责人: Justin Cobb Paglino
• 金额: $ 11.74万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-04-01 至 2018-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9056439
• 关键词: Achievement; Animals; Antibodies; Antigen-Presenting Cells; Award; Blocking Antibodies; Brain; C57BL/6 Mouse; CD8B1 gene; CTLA4 gene; Cancer Immunology Science; Capsid; Cell secretion; Cells; Clinic; Clinical; Clinical Pathology; Combined Modality Therapy; Complementary DNA; Cytolysis; Cytotoxic T-Lymphocyte-Associated Protein 4; Data; Dependence; Development; Development Plans; Diagnosis; Disease; Engineering; Epitopes; Faculty; Feedback; Genetic; Genome; Glioblastoma; Glioma; Goals; Granulocyte-Macrophage Colony-Stimulating Factor; Growth; Heterophile Antigens; Human; Immune; Immune Tolerance; Immune response; Immune system; Immunity; Immunobiology; Immunocompetent; Immunocompromised Host; Immunotherapeutic agent; Immunotherapy; In Vitro; Individual; Infection; Injection of therapeutic agent; Interleukin-12; Journals; K-Series Research Career Programs; Knowledge; Laboratories; Libraries; Lytic; Malignant Glioma; Malignant Neoplasms; Measures; Mediating; Mediator of activation protein; Medical; Medicine; Mentors; Mentorship; Modeling; Molecular; Molecular Mimicry; Molecular Virology; Mus; Neuroglia; Neurosurgeon; Oncolytic; Oncolytic viruses; Operative Surgical Procedures; Outcome; PPBP gene; Paper; Parvovirus; Parvovirus Infections; Patients; Positioning Attribute; Prostate; Prostatic Neoplasms; Proteins; Publications; Publishing; Radiation; Recombinant adeno-associated virus (rAAV); Recombinants; Regulatory T-Lymphocyte; Reporting; Research; Research Personnel; Residencies; Resistance; Safety; Signal Transduction; Splenocyte; Structure; Survival Analysis; T-Cell Depletion; T-Cell Development; T-Lymphocyte; Techniques; Testing; Therapeutic; Time; TimeLine; Training; Transgenes; Tropism; Tumor Antigens; Tumor Biology; Tumor Immunity; United States; Vesicular stomatitis Indiana virus; Viral; Viral Vector; Virus; Virus Replication; Yale Cancer Center; adeno-associated viral vector; base; cancer immunotherapy; career; career development; chemotherapy; combinatorial; cytokine; design; experience; implantation; interest; killings; knowledge base; meetings; member; mouse model; neoplastic cell; neuro-oncology; neurosurgery; novel; novel strategies; novel therapeutics; oncology program; oncolysis; oncolytic Vesicular Stomatitis Virus; preference; professor; programs; research and development; responsible research conduct; sarcoma; skills; success; symposium; tenure track; tumor; tumor microenvironment; vector; virology

DESCRIPTION (provided by applicant): This application describes a plan to develop my career through fully-time research, structured mentoring, didactics, seminars, training in the responsible conduct of research, presentation at conferences, and specific target dates for the achievement of goals that include completion and publication of data described in my specific aims, committee guidance for moving forward to a tenure-track faculty position, and submission of an R01 proposal. The research strategy describes our approach to attempting to cure syngeneic orthotopic malignant glioma using an oncolytic parovirus combined with therapies that we hypothesize will synergize with parvovirus therapy because they operate by different mechanisms. The use of parvovirus in an orthotopic syngeneic glioma model is novel, as is the combination of parvovirus with the chosen complementary approaches, which are all of significant promise or demonstrated efficacy. I am well prepared to pursue the goals outlined in this application. My medical training and residency in clinical pathology give me a comprehensive knowledge base for understanding disease and its treatment. My graduate training in molecular parvovirology, under Dr. Peter Tattersall (referee) in the department of Laboratory Medicine at Yale equipped me with skills in a variety of molecular virology techniques, developed my ability to design an effective experimental approach, and resulted in two first-author publications in Virology on the tropism of oncolytic parvoviruses. In my subsequent training with Dr. Anthony van den Pol, here the primary mentor, I have gained facility in using oncolytic vesicular stomatitis virus (VSV), designing and generating a number of recombinant adeno-associated virus (AAV) vectors, and studying tumor oncolysis in immunocompromised mouse models. In 2011, I published a first-author article in the Journal of Virology on the applicability of VSV to treatment of human sarcomas, in which we demonstrate that in some sarcomas, constitutive innate immune signaling poses a barrier to VSV replication; I found several ways in which to enhance viral targeting to otherwise resistant sarcomas. This year, I published another first-author paper in the Journal of Virology on parvoviral oncolysis of gliomas in vitro and in immunocompromised mice. My research interest from the start of my training has been to employ viruses as cancer therapeutics. To me, the most exciting developments in this field have been the discovery of means by which viruses can stimulate an effective, even curative, anti-tumor immune response. Although I have knowledge of immunobiology and cancer immunotherapy, I seek to understand it on a level that will allow me to make needed advances in this field. Therefore I am applying for this career development award to receive training and undertake research with an emphasis on immunobiology, particularly the use of viruses and viral vectors as immunotherapeutics. The career development plan describes meeting monthly with mentors one-on-one, and quarterly with the entire committee, to whom my research and career development activities will be presented for feedback. My situation within the department of neurosurgery and the experience of my primary mentor make glioma the ideal target disease for my research. Dr. van den Pol has published reports on the use of a number viruses to target glioma, including our most recent report investigating parvoviruses. Co-mentor Dr. Joseph Piepmeier is a practicing neurosurgeon, director of the Surgical Neuro-Oncology Program, and was editor-in-chief of the Journal of Neuro-Oncology for eight years. I will attend tumor-board with Dr. Piepmeier. Co- mentor Dr. Daniel DiMaio, professor of Genetics and scientific director of the Yale Cancer Center, will be valuable as a mentor given his expertise in both molecular virology and tumor biology. Co-mentorship by Dr. Alfred Bothwell, Professor of Immunobiology, is a critical feature of this application. A member of the Yale Cancer Center's Cancer-Immunology Program, and Director of Graduate Studies, his immunobiology expertise combined with an excellent track record of mentoring will be indispensable to me as I endeavor to broaden my research into his field. Other important components of the career development plan include an immunobiology course, an advanced course in T-cell development, attending relevant research and clinical seminars, annual presentation at conferences, and a timeline for achievement of research and career goals. Our approach is to most closely mimic clinical glioma by using an orthotopic syngeneic immunocompetent model, and we have identified two glioma lines syngeneic to C57BL/6 mice that are susceptible to lytic growth of parvovirus MVMp. The MVMp genome is 80% identical to that of parvovirus LuIII, the virus we recently reported to grow optimally in human gliomas, however the human species preference of LuIII precludes its use in syngenic mouse models. Evidence supports our view that parvoviral oncolysis enhances release of tumor antigens in an infection-stimulated microenvironment, thereby priming an anti- tumor immune response. Central to my approach is the perspective that optimal efficacy depends on a combination of therapeutics that coordinately promote an anti-tumor immune response by different mechanisms. Therefore, I test the hypothesis that combinatorial strategies, in which oncolytic parvovirus infection is combined with different complementary immune stimulatory treatments, will constitute effective immunotherapy for glioma. In Aim 1 I test three immunostimulatory antibodies. In Aim 2 I test two parvoviral vectors expressing cytokines. In Aim 3 I test an 'altere self epitope library' engineered into parvoviral vector. I look forward to the research and trainin activities described herein.

描述（由申请人提供）：本申请描述了一项通过全职研究、结构化指导、教学、研讨会、负责任的研究行为培训、会议演讲和实现目标的具体目标日期来发展我的职业生涯的计划，这些目标包括完成和发布我的具体目标中描述的数据、委员会对迈向终身教职的指导，以及提交R01提案。该研究策略描述了我们尝试使用溶瘤性小病毒结合治疗方法来治疗同源原位恶性胶质瘤的方法，我们假设这些方法将与小病毒治疗协同作用，因为它们通过不同的机制起作用。在同种胶质瘤模型中使用细小病毒是新颖的，将细小病毒与所选择的补充方法相结合也是新颖的，这些方法都有重要的前景或已证明有效。我已经做好了充分的准备去追求这份申请中列出的目标。我在临床病理学方面的医学培训和住院实习为我了解疾病及其治疗提供了全面的知识基础。在耶鲁大学检验医学系Peter Tattersall博士（审稿人）的指导下，我在分子小病毒学方面进行了研究生培训，掌握了各种分子病毒学技术，培养了我设计有效实验方法的能力，并在《病毒学》上发表了两篇关于溶瘤性小病毒趋向性的第一作者论文。在随后跟随主要导师Anthony van den Pol博士的培训中，我掌握了使用溶瘤性水泡性口炎病毒（VSV）的技能，设计和生成了许多重组腺相关病毒（AAV）载体，并在免疫功能低下的小鼠模型中研究肿瘤溶瘤。2011年，我在《病毒学杂志》（Journal of Virology）上发表了一篇关于VSV在治疗人类肉瘤中的适用性的第一作者文章，其中我们证明了在一些肉瘤中，组成性先天免疫信号对VSV的复制构成了障碍；我发现了几种方法来增强病毒靶向性，以抵抗肉瘤。今年，我在《病毒学杂志》（Journal of Virology）上发表了另一篇第一作者论文，内容是在体外和免疫功能低下的小鼠中对胶质瘤进行细小病毒溶瘤。我的研究兴趣从一开始就是利用病毒来治疗癌症。对我来说，这个领域最令人兴奋的发展是发现了病毒可以刺激有效的、甚至是治愈的抗肿瘤免疫反应的方法。虽然我有免疫生物学和癌症免疫治疗的知识，但我试图在一定程度上理解它，这将使我在这一领域取得必要的进展。因此，我申请这个职业发展奖，接受培训，并从事研究，重点是免疫生物学，特别是使用病毒和病毒载体作为免疫治疗。职业发展计划描述了每月与导师一对一会面，每季度与整个委员会会面，我的研究和职业发展活动将向他们汇报，以获得反馈。我在神经外科的情况和我的主要导师的经验使胶质瘤成为我理想的研究目标疾病。van den Pol博士发表了关于使用多种病毒靶向胶质瘤的报告，包括我们最近研究细小病毒的报告。共同导师Joseph Piepmeier博士是一名执业神经外科医生，外科神经肿瘤学项目主任，并担任《神经肿瘤学杂志》主编八年。我会和皮普梅尔医生一起参加肿瘤委员会。共同导师Daniel DiMaio博士是耶鲁大学癌症中心遗传学教授和科学主任，鉴于他在分子病毒学和肿瘤生物学方面的专业知识，他将是一位有价值的导师。由免疫生物学教授Alfred Bothwell博士共同指导是该应用程序的关键特征。作为耶鲁大学癌症中心癌症免疫学项目的成员和研究生研究主任，他的免疫生物学专业知识和出色的指导记录对我来说是不可或缺的，因为我努力将我的研究扩展到他的领域。职业发展计划的其他重要组成部分包括免疫生物学课程，t细胞发育高级课程，参加相关研究和临床研讨会，年度会议报告，以及实现研究和职业目标的时间表。我们的方法是通过使用原位同源免疫活性模型来最接近地模拟临床胶质瘤，我们已经确定了两个与C57BL/6小鼠同源的胶质瘤系，它们对细小病毒MVMp的裂解生长敏感。MVMp基因组与我们最近报道的在人类胶质瘤中生长最佳的细小病毒LuIII的基因组有80%相同，然而人类对LuIII的物种偏好使其无法在同基因小鼠模型中使用。证据支持我们的观点，即在感染刺激的微环境中，细小病毒溶瘤促进肿瘤抗原的释放，从而引发抗肿瘤免疫反应。我的方法的核心观点是，最佳疗效取决于通过不同机制协调促进抗肿瘤免疫反应的治疗方法的组合。因此，我验证了溶瘤细小病毒感染与不同互补免疫刺激治疗相结合的组合策略将构成对胶质瘤有效的免疫治疗的假设。在Aim 1中，我测试了三种免疫刺激抗体。在Aim 2中，我测试了两个表达细胞因子的细小病毒载体。在Aim 3中，我测试了一个改造成细小病毒载体的“改变自身表位库”。我期待着这里所描述的研究和培训活动。