Development and validation of a genetically engineered model of neurofibromatosis type 2 to facilitate discovery of neurotherapeutics

开发和验证 2 型神经纤维瘤病基因工程模型，以促进神经治疗药物的发现

• 批准号: 9375911
• 负责人: JOSEPH KISSIL
• 金额: $ 47.72万
• 依托单位: SCRIPPS FLORIDA
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-08-01 至 2019-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9375911
• 关键词: Acoustic Neuroma; Affect; Alleles; Anatomy; Animal Model; Animals; Benchmarking; Bilateral; Biology; Bioluminescence; Breeding; Clinical Trials; Cranial Nerves; Development; Disease; Drug Evaluation; Energy Transfer; Enterobacteria phage P1 Cre recombinase; Enzymes; Ependymoma; FRAP1 gene; Genes; Genetic Engineering; Genetic Recombination; Genetically Engineered Mouse; Genotype; Goals; Growth and Development function; Histologic; Imaging technology; Inherited; Intervention; Knockout Mice; Light; Link; Loss of Heterozygosity; Luciferases; MAP Kinase Gene; Mediating; Modeling; Molecular; Monitor; Monomeric GTP-Binding Proteins; Morbidity - disease rate; Mus; Mutant Strains Mice; Nervous system structure; Neurilemmoma; Neurofibromatosis 2; Neurofibromin 2; Operative Surgical Procedures; Patients; Penetration; Pharmaceutical Preparations; Pharmacotherapy; Phase; Proteins; Radiation; Radiation therapy; Regulation; Reporter; Reporting; Research Personnel; Schwann Cells; Shrimp; Signal Pathway; Signal Transduction; Stem cells; Symptoms; Technology; Testing; Therapeutic; Time; Tissues; Transgenic Mice; Transgenic Organisms; Tumor Suppressor Genes; Tumor Volume; Validation; Variant; absorption; aged; bevacizumab; cohort; deep ocean; drug candidate; drug testing; effective therapy; experience; histopathological examination; human disease; improved; in vivo; in vivo optical imaging; meningioma; mortality; mouse model; neoplastic cell; novel; periostin; pre-clinical; promoter; response; serial imaging; spatiotemporal; therapeutic target; treatment effect; treatment response; tumor; tumor growth

Project Summary: Neurofibromatosis type 2 (NF2) is a dominantly inherited autosomal disease (affecting 1 in 30,000) that is attributed to loss-of-heterozygosity (LOH) of the NF2 gene. By far the most common manifestation of the disease is the development of schwannomas of the 8th cranial nerve. Although our understanding of the molecular mechanisms underlying NF2 has significantly improved over the past 2 decades, and a number of potential therapeutic targets have been identified, viable treatment options are still lacking for this disease. Current treatment options are temporary anti-symptomatic interventions that are limited to radiation and/or surgery and associated with severe morbidity. Clearly, there is an urgent need to develop therapeutic options for NF2 patients. Currently, one of the main obstacles towards the development of effective drug treatments for NF2 is the lack of appropriate animal models to enable pre-clinical testing of drug candidates. To overcome this, we propose to develop a GEMM (Genetically Engineered Mouse Model) of NF2 that accurately reflects the biology of schwannoma development in patients, to incorporate a transgenic reporter allele allowing the temporal evaluation of drug candidate efficacy in an accurate and consistent manner. Towards this goal we will employ a recently described NF2 GEMM in which both Nf2 alleles are inactivated in Schwann cell progenitors, by crossing Nf2 conditional knockout mice to transgenic mice carrying a Cre-recombinase allele driven by the Periostin promoter. These mice will then be crossed to a reporter strain we developed, in which a novel bioluminescence resonance energy transfer (BRET) reporter allele can be conditionally turned on by Cre- mediated recombination. This newly developed reporter overcomes limitations previously experienced in animal models, such as a requirement for externally provided excitation light and limited penetration due to tissue absorption, by using a fusion of enhanced GFP to an enhanced variant of luciferase. The intra-molecular BRET between these proteins generates the brightest bioluminescent signal known to date and improves spatiotemporal monitoring of small numbers of tumor cells using in vivo optical imaging. In the R21 phase we will develop and internally validate this new NF2 GEMM. In the R33 phase we will determine the predicitive validity of the GEMM by testing it with 2 drugs previously shown to elicit an anti-tumor response in transplantable models of NF2. This will permit us to follow tumor growth and response to treatment, over an extended time period, in a GEMM that closely reflects the human disease.

项目概要： 2型神经纤维瘤病（NF 2）是一种显性遗传的常染色体疾病（影响1/30，000）， 这归因于NF 2基因的杂合性缺失（洛）。到目前为止， 疾病是第八脑神经的神经鞘瘤的发展。尽管我们对 在过去的20年里，NF 2的分子机制得到了显著的改善， 尽管已经确定了潜在的治疗靶点，但仍然缺乏针对这种疾病的可行的治疗选择。 目前的治疗选择是暂时的对症干预，仅限于放射和/或 手术和严重并发症。显然，迫切需要开发治疗方案 NF 2患者 目前，开发针对NF 2的有效药物治疗的主要障碍之一是缺乏针对NF 2的有效药物治疗。 适当的动物模型，使候选药物的临床前测试。为了克服这一点，我们建议 开发一种NF 2的GEMM（基因工程小鼠模型），准确反映了 神经鞘瘤发展的患者，纳入转基因报告等位基因，允许颞叶神经鞘瘤， 以准确和一致的方式评价候选药物的疗效。为了实现这一目标，我们将 最近描述的Nf 2 GEMM，其中两个Nf 2等位基因在雪旺细胞祖细胞中被失活， 将Nf 2条件性敲除小鼠与携带Cre-重组酶等位基因的转基因小鼠杂交，所述Cre-重组酶等位基因由Nf 2基因驱动。 骨膜蛋白启动子。然后这些小鼠将与我们开发的一种报告菌株杂交， 生物发光共振能量转移（BRET）报告等位基因可以通过Cre- 介导的重组这种新开发的报告器克服了以前在 动物模型，如需要外部提供的激发光和有限的穿透， 组织吸收，通过使用增强的GFP与增强的荧光素酶变体的融合。分子内 这些蛋白质之间的BRET产生迄今为止已知的最亮的生物发光信号， 使用体内光学成像对少量肿瘤细胞进行时空监测。在R21阶段，我们 将开发和内部验证这种新的NF 2 GEMM。在R33阶段，我们将确定 GEMM的有效性，通过用2种先前显示可引起抗肿瘤反应的药物进行测试， NF 2的可移植模型。这将使我们能够跟踪肿瘤生长和对治疗的反应，超过一个月。 延长的时间段，在GEMM，密切反映人类疾病。