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: 8634751
• 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/8634751
• 关键词: Achievement; Am 80; 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; 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; 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; oncology program; oncolysis; preference; professor; programs; research and development; responsible research conduct; sarcoma; skills; success; symposium; 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）：本申请描述了一个通过全职研究、结构化指导、教学、研讨会、负责任的研究行为培训、会议演讲以及实现目标的具体目标日期来发展我的职业生涯的计划其中包括完成和发布我的具体目标中描述的数据、晋升终身教授职位的委员会指导以及提交 R01 提案。该研究策略描述了我们尝试使用溶瘤细小病毒与细小病毒疗法相结合来治愈同基因原位恶性胶质瘤的方法，我们假设这些疗法将与细小病毒疗法产生协同作用，因为它们通过不同的机制发挥作用。细小病毒在原位同基因神经胶质瘤模型中的使用是新颖的，细小病毒与所选补充方法的组合也是新颖的，这些方法都具有重大前景或已证明有效。 我已做好充分准备来实现本申请中概述的目标。我的医学培训和临床病理学住院医师培训为我了解疾病及其治疗提供了全面的知识基础。我在耶鲁大学实验医学系 Peter Tattersall 博士（裁判）的指导下进行分子细小病毒学研究生培训，使我掌握了各种分子病毒学技术的技能，培养了我设计有效实验方法的能力，并取得了两项第一-作者在病毒学上发表了有关溶瘤细小病毒趋向性的文章。在随后与主要导师 Anthony van den Pol 博士一起进行的培训中，我在使用溶瘤性水泡性口炎病毒 (VSV)、设计和生成多种重组腺相关病毒 (AAV) 载体以及研究肿瘤方面获得了一定的技巧。免疫受损小鼠模型中的溶瘤作用。 2011 年，我在《病毒学杂志》上发表了一篇关于 VSV 治疗人类肉瘤的适用性的第一作者文章，其中我们证明，在某些肉瘤中，组成型先天免疫信号传导对 VSV 复制构成了障碍；我发现了几种增强病毒对耐药肉瘤的靶向作用的方法。今年，我在《病毒学杂志》上发表了另一篇第一作者论文，内容涉及体外和免疫功能低下小鼠体内神经胶质瘤的细小病毒溶瘤作用。从培训之初起，我的研究兴趣就是利用病毒作为癌症疗法。对我来说，这个领域最令人兴奋的进展是发现病毒可以刺激有效的、甚至是治愈性的抗肿瘤免疫反应的方法。尽管我了解免疫生物学和癌症免疫治疗的知识，但我寻求在一定程度上理解它，以便我能够在该领域取得所需的进展。因此，我正在申请这个职业发展奖，以接受培训并进行免疫生物学研究，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的会议，每季度与整个委员会的会议，我的研究和职业发展活动将向他们提出反馈。我在神经外科的情况和我的主要导师的经验使神经胶质瘤成为我研究的理想目标疾病。 van den Pol 博士发表了关于使用多种病毒来靶向神经胶质瘤的报告，包括我们最近研究细小病毒的报告。共同导师 Joseph Piepmeier 博士是一位执业神经外科医生、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。我将与 Piepmeier 博士一起参加肿瘤委员会。导师 Daniel DiMaio 博士是耶鲁大学癌症中心的遗传学教授兼科学主任，鉴于他在分子病毒学和肿瘤生物学方面的专业知识，他将成为有价值的导师。免疫生物学教授 Alfred Bothwell 博士的共同指导是该应用程序的一个关键特征。作为耶鲁大学癌症中心癌症免疫学项目的成员和研究生院主任，他的免疫生物学专业知识加上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、年度会议演讲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基因免疫活性模型来最接近地模拟临床神经胶质瘤，并且我们已经确定了两个与 C57BL/6 小鼠同基因的神经胶质瘤系，它们对细小病毒 MVMp 的裂解生长敏感。 MVMp 基因组与细小病毒 LuIII 的基因组有 80% 相同，我们最近报道该病毒在人类神经胶质瘤中生长最佳，然而人类对 LuIII 的物种偏好阻碍了其在同基因小鼠模型中的使用。有证据支持我们的观点，即细小病毒溶瘤增强了感染刺激的微环境中肿瘤抗原的释放，从而引发抗肿瘤免疫反应。我的方法的核心是这样的观点：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组合。因此，我检验了这样的假设：溶瘤细小病毒感染与不同的互补免疫刺激治疗相结合的组合策略将构成神经胶质瘤的有效免疫疗法。在目标 1 中，我测试了三种免疫刺激抗体。在目标 2 中，我测试了两种表达细胞因子的细小病毒载体。在目标 3 中，我测试了一个被工程化到细小病毒载体中的“改变自身表位库”。我期待本文所述的研究和培训活动。