Mechanisms of Caspase-1 Mediated Schwannoma Regression

Caspase-1 介导神经鞘瘤消退的机制

• 批准号: 8703827
• 负责人: GARY JAY BRENNER
• 金额: $ 36.02万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-30 至 2017-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8703827
• 关键词: Animals; Apoptosis; Apoptotic; Benign; Biochemical; Biological; Biological Assay; Biology; Bystander Effect; CASP1 gene; Caspase; Caspase-1; Cause of Death; Cell Death; Cell Lineage; Cells; Cessation of life; Clinical; Code; Culture Media; DNA; Data; Dependovirus; Development; Disease; Disease Management; Elements; Environment; Genes; Goals; Grant; Hereditary Disease; Human; Image; Immune; Immune response; Immunocompetent; Implant; In Vitro; Individual; Infection; Inflammatory; Injection of therapeutic agent; Investigation; Lead; Lesion; Location; Magnetic Resonance Imaging; Manuscripts; Mediating; Medical; Modeling; Molecular; Monitor; Mus; Myelin Sheath; NF1 gene; Natural Killer Cells; Neoplasms; Nerve; Neurilemmoma; Neurofibromatoses; Neurofibromatosis 2; Neurons; Non-Malignant; Nude Mice; Pathway interactions; Patients; Peripheral Nerves; Peripheral Nervous System Neoplasms; Primary Neoplasm; Publishing; Research; Resistance to infection; Schwann Cells; Schwannomatosis; Stream; Techniques; Testing; Therapeutic; Therapeutic Effect; Time; Toxic effect; Treatment Efficacy; Tumor Volume; Tumor-Derived; Viral Vector; Virus; adaptive immunity; adeno-associated viral vector; cytokine; disability; gene therapy clinical trial; implantation; in vivo; innovation; killings; myelination; neoplastic cell; neurotoxicity; novel; novel therapeutic intervention; novel therapeutics; preclinical study; promoter; protein complex; response; sciatic nerve; treatment strategy; tumor; tumor growth; vector; vector control

DESCRIPTION (provided by applicant): While schwannoma tumors are typically benign (non-malignant) they can have devastating consequences for patients. As for an individual patient there is often no efficacious treatment. Thus, schwannoma therapy represents a major unmet clinical need. We have developed a new therapeutic approach to these nerve-associated tumors that involves intra-tumoral injection of an adeno-associated virus (AAV) vector carrying the pro-apoptotic gene caspase-1 (ICE) under a Schwann-cell specific promoter (P0); the vector is denoted AAV-P0-ICE. Our pre-clinical studies have shown a remarkable ability of the AAV-P0- ICE vector to produce a prolonged reduction in tumor volume with no nerve damage (manuscript submitted). Our preliminary data suggests that AAV-PO-ICE not only directly kills infected caspase-1-expressing tumor cells (presumably by inducing apoptosis), but also kills non-infected tumor cells through a "bystander" effect. It is hypothesize that this bystander killin is mediated by both tumor- and host-specific mechanisms that involve humoral- and cell-mediated pathways. The overriding goal is to investigate the biology of AAV-P0-ICE mediated tumor destruction in the human and mouse schwannoma models we have developed in nude and immunocompetent mice, respectively, and to identify the molecular and cellular pathways mediating the killing of non-vector-infected tumor cells that is associated with apoptotic death of infected tumor cells. In brief, the specific aims are to 1) investigate the parameters of tumor regression in two schwannoma models in mice - a human schwannoma in nude mice and a mouse schwannoma in immune competent animals, 2) investigate whether caspase-1 bystander killing involves apoptotic death of uninfected cells, and 3) analyze whether host immune mechanisms are involved in AAV-P0-ICE initiated bystander killing. In vivo tumor growth will be monitored via bioluminescent imaging and MRI for the human and mouse schwannoma models, respectively; both techniques allow serial assessment over time in a given animal. A variety of biochemical and cell biological techniques will be used to determine whether caspase expressing tumor cells produce factors - both humoral and microvesicle contained - that can be transferred to and cause apoptotic death of uninfected tumor cells. Histological analysis and a variety of fundamental immunological assays will be used to investigate host mechanisms of bystander killing after AAV-P0-ICE injection. This application is novel and innovative in that it utilizes the first viral vector we know of that is restrictively expressed in Schwann-lineage cells hence allowing targeting of schwannomas without neurotoxicity, and proposes investigation of the pathways - tumor and host specific - which underlie induction of bystander tumor killing. Mechanistic understanding of the extensive and specific tumor killing induced by AAV-P0-ICE injection into schwannomas has the potential lead to novel treatment strategies for NF1, NF2 and schwannomatosis, as well as, a variety of other neoplasms.

描述（由申请人提供）：虽然切旺纳马瘤肿瘤通常是良性（非恶性），但它们可能会对患者产生毁灭性的后果。至于个体患者，通常没有有效的治疗方法。因此，切瓦纳瘤治疗代表了主要的未满足的临床需求。我们已经为这些神经相关的肿瘤开发了一种新的治疗方法，涉及在schwann-cell特异性启动子（p0）下载有促凋亡基因caspase-1（ice）的腺相关病毒（AAV）载体（AAV）载体；向量表示AAV-P0冰。我们的临床前研究表明，AAV-P0冰载体具有显着的能力，可以长期减少肿瘤体积而没有神经损伤（手稿提交）。 我们的初步数据表明，AAV-PO-ICE不仅直接杀死了受感染的表达caspase-1表达肿瘤细胞（大概是通过诱导凋亡），而且还通过“旁观者”效应杀死了未感染的肿瘤细胞。假设该旁观者基林是由涉及体液和细胞介导的途径的肿瘤和宿主特异性机制介导的。压倒性的目标是研究我们在裸体和免疫能力的小鼠中开发的人和小鼠造型模型中AAV-P0冰介导的肿瘤破坏的生物学，并确定介导与非载体感染的肿瘤细胞杀死的分子和细胞途径，该途径与非载体的肿瘤细胞杀死相关。 感染的肿瘤细胞。 简而言之，具体目的是1）研究小鼠两种切氏瘤模型中肿瘤回归的参数 - 裸鼠中的人类造口瘤和免疫能力动物中的小鼠造型症的参数，2）研究CASPase-1旁观者杀害杀害是否涉及未感染细胞的凋亡死亡涉及宿主免疫机构的杀人剂，是否涉及杀人机构。人体和小鼠造型瘤模型将通过生物发光成像和MRI来监测体内肿瘤的生长。这两种技术都允许在给定动物中随着时间的流逝进行连续评估。各种生化和细胞生物学技术将用于确定表达肿瘤细胞的胱天蛋白酶是否产生可能会转移到未感染肿瘤细胞的凋亡死亡的因素（包括体液和微孔）。组织学分析和各种基本免疫学测定将用于研究AAV-P0冰注射后旁观者杀死的宿主机制。 该应用是新颖和创新的，因为它利用了我们所知道的第一个病毒载体在schwann -linege细胞中限制性表达，因此允许靶向无神经毒性的schwannomas，并提出对途径的研究 - 肿瘤和宿主特异性 - 这是对旁观者降落杀伤的诱导。 AAV-P0-ICE注入Schwannomas引起的广泛而特定的肿瘤杀伤的机械理解可能导致NF1，NF2和Schwannomatosis的新型治疗策略，以及其他各种肿瘤。