Enhancing Delivery and Regulating Gene Expression in Oncolytic Vaccinia Vectors

增强溶瘤痘苗载体的递送和调节基因表达

• 批准号: 8019447
• 负责人: Stephen H Thorne
• 金额: $ 30.49万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-04-01 至 2015-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8019447
• 关键词: Address; Adverse reactions; Animal Model; Antibodies; Area; Binding; Biological Products; Biological Response Modifier Therapy; Blood Vessels; Bystander Effect; Cancer Patient; Cell Therapy; Cells; Clinical; Clinical Trials Design; Data; Development; Disease; Drug resistance; Early treatment; Effectiveness; Environment; Epitopes; Excision; Exposure to; Extracellular Matrix; Future; Gene Expression; Genes; Immune; Immune Targeting; Immune response; Immunotherapy; In Situ; Infection prevention; Killer Cells; Lead; Malignant Neoplasms; Mediating; Microbubbles; Mus; Mutation; Necrosis; Oncolytic; Oncolytic viruses; Outcome; Patients; Phase; Pre-Clinical Model; Production; Proteins; Protocols documentation; Regulation; Relapse; Safety; Smallpox; Testing; Therapeutic; Transgenes; Ultrasonography; Universities; Vaccinated; Vaccination; Vaccinia; Vaccinia virus; Vaccinium; Vascular System; Vascular blood supply; Viral; Viral Genes; Viral Load result; Viral Proteins; Viral Vector; Virus; Virus Diseases; Work; alternative treatment; base; cancer cell; cancer therapy; cytokine; design; experience; gene function; immune clearance; improved; neutralizing antibody; oncolytic vector; particle; passive transport; pre-clinical; public health relevance; research clinical testing; response; targeted delivery; therapeutic transgene; traditional therapy; transgene expression; tumor; tumor eradication; vaccinia virus vector; vector

DESCRIPTION (provided by applicant): We have previously designed and implemented a variety of oncolytic vectors based on vaccinia virus, developing a panel of gene-deleted and tumor-targeting strains that have displayed promising pre- clinical and early clinical results. We have also pioneered the use of cell-based carrier vehicles to more efficiently deliver oncolytic virus strains to their tumor targets. In particular we demonstrated that Cytokine Induced Killer (CIK) cells could not only act as effective delivery vehicles for vaccinia strains, but could also synergize with the virus to increase their anti-tumor effects. Despite these recent advances, these therapies have still not reached their full potential. In this proposal we have identified key limitations to the future development of these biological therapies, including the requirement to overcome anti-viral immune responses in order to efficiently repeat treat with the same therapy; and the need for both added potency and improved safety within these vectors. We look to address these in three main aims: in the first two aims we will focus on improving the systemic delivery of the viral vector to the tumor, especially in pre-immunized animal models and look to implement strategies to even take advantage of the anti-viral immune response; in the final aim we will examine the advantages of placing different transgene or essential viral gene products under the control of externally regulated protein stability domains. A total of three complimentary approaches will be applied to enhance viral delivery in the face of an anti-viral immune response, focusing on evading circulating neutralizing antibody; (i) epitope swapping or epitope removal will be used to reduce antibody recognition, (ii) binding of viral particles to targeted microbubbles, followed by ultrasound mediated release within the tumor environment and (iii) use of cell carriers to deliver virus to the tumor, particularly incorporating viral strains with mutations that increase the levels of the 'stealth' or enveloped form of the virus produced in situ. In the final aim we will further define and incorporate previously demonstrated advantages of externally regulating the function of different therapeutic transgenes expressed from the viral vectors. Alternatively, we will regulate essential viral gene expression, so providing a shut-off switch within the vector. The overall objectives of this proposal are therefore to overcome some of the major limitations of oncolytic viral therapy, so improving its clinical potential and providing realistic alternative treatment options for patients with some of the hardest to treat cancers, including relapsed, drug-resistant and disseminated disease. Data produced in this proposal will be directly applied to the protocol design of clinical trials under development at the University of Pittsburgh, and so we expect it to directly lead to patient benefits. PUBLIC HEALTH RELEVANCE: This overall aim of this proposal is to enhance the application of oncolytic (replication selective) viruses. This will primarily involve improving the systemic delivery of these vectors, both in naove and previously immunized hosts. In addition we will incorporate regulated transgene function to improve the effectiveness of these therapies and we will look to integrate the oncolytic viruses with immune cell therapies. The focus of this proposal will not only be on improving the biological agents as single therapies, but also on how to combine them such that the natural interactions between viruses and immune cell therapies can be utilized to enhance the therapeutic outcome.

描述（由申请人提供）：我们以前已经设计并实施了基于离子病毒的各种溶瘤载体，开发了一组基因删除和涉及肿瘤的靶向菌株，这些菌株显示出有希望的临床前和早期临床结果。我们还开创了基于细胞的载体车辆的使用，以更有效地将溶瘤病毒菌株传递到其肿瘤靶标。特别是我们证明了细胞因子诱导的杀伤（CIK）细胞不仅可以充当有效的离发疫苗菌株的递送车，而且还可以与病毒协同作用以增加其抗肿瘤作用。尽管这些最近的进步，这些疗法仍未发挥其全部潜力。在该提案中，我们确定了这些生物疗法未来发展的关键局限性，包括要克服抗病毒免疫反应以进行相同疗法有效重复治疗的要求；以及在这些向量中增加效力和提高安全性的需求。我们打算在三个主要目的中解决这些问题：在前两个目标中，我们将重点放在改善病毒载体向肿瘤的系统性输送上，尤其是在预免疫模型中，并寻求实施策略，甚至利用抗病毒免疫反应；在最终目的中，我们将研究将不同的转基因或必需病毒基因产物置于外部调节蛋白质稳定性域的控制之下的优势。面对抗病毒免疫反应，总共将采用三种免费方法来增强病毒递送，重点是逃避循环中和中和抗体。 （i）表位交换或去除表位将用于减少抗体的识别，（ii）病毒颗粒与靶向微泡的结合，然后在肿瘤环境中进行超声介导的释放，（iii）使用细胞载体将病毒与肿瘤传递到肿瘤中，尤其是将病毒株与``''steal of theeble of theeble of theebles of shopery ofere nebleseples'prode nepresseal'profence conseption'profence'insovers of inseal''或endere hosende in neblesefence'propence of inseal''的水平。在最终目的中，我们将进一步定义并结合先前证明的外部调节从病毒载体表达的不同治疗转基因功能的优势。另外，我们将调节必需的病毒基因表达，因此在向量内提供关闭开关。因此，该提案的总体目标是克服癌病毒疗法的一些主要局限性，从而提高其临床潜力，并为患有最难治疗癌症的患者提供现实的替代治疗选择，包括复发，耐药和散布疾病。本提案中生产的数据将直接应用于匹兹堡大学正在开发的临床试验方案设计，因此我们希望它直接带来患者益处。 公共卫生相关性：该提案的总体目的是增强溶瘤（复制选择性）病毒的应用。这将主要涉及改善在NAOVE和以前免疫的宿主中的这些向量的系统性传递。此外，我们将结合受调节的转基因功能以提高这些疗法的有效性，我们将寻求将溶瘤病毒与免疫细胞疗法相结合。该提案的重点不仅将放在改善生物学剂作为单一疗法上，而且还放在如何结合它们上，以便可以利用病毒与免疫细胞疗法之间的自然相互作用来增强治疗结果。