ConProject-001

ConProject-001

• 批准号: 9981226
• 负责人: JOSEPH KISSIL
• 金额: $ 11.76万
• 依托单位: SCRIPPS FLORIDA
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-01 至 2020-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9981226
• 关键词: Acoustic Neuroma; Affect; Alleles; Anatomy; Animal Model; Animals; Antitumor Response; 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; 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; Preclinical Testing; 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; effective therapy; experience; histopathological examination; human disease; improved; in vivo; in vivo optical imaging; meningioma; mortality; neoplastic cell; novel; periostin; promoter; serial imaging; spatiotemporal; therapeutic target; transplant model; treatment effect; treatment response; tumor; tumor growth

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 Cremediated 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.

神经纤维瘤病2型(NF2)是一种显性遗传性常染色体疾病(每30,000人中就有1人患病)，即 归因于NF2基因的杂合性缺失(LOH)。迄今为止，最常见的表现为 本病是第8脑神经的神经鞘瘤发展而来。虽然我们对这一现象的理解 在过去的20年里，NF2的分子机制已经有了显著的改善，一些 潜在的治疗靶点已经确定，但对这种疾病仍然缺乏可行的治疗选择。 目前的治疗方案是仅限于放射和/或治疗的临时性抗症状干预 手术，并伴随着严重的发病率。显然，迫切需要开发治疗方案。 适用于NF2患者。 目前，开发针对NF2的有效药物治疗的主要障碍之一是缺乏 建立合适的动物模型，以便对候选药物进行临床前测试。为了克服这一点，我们建议 开发一种准确反映NF2生物学特性的NF2基因工程小鼠模型 神经鞘瘤患者的发展，纳入转基因报告等位基因允许颞叶 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，我们将采用 最近描述的一种NF2 GEMM，其中两个NF2等位基因在雪旺细胞祖细胞中都被灭活，通过 将Nf2条件基因敲除小鼠与携带Cre重组酶等位基因的转基因小鼠进行杂交，该等位基因由 周膜蛋白启动子。然后，这些小鼠将被杂交到我们开发的一种报告菌株，在这种菌株中，一种新的 生物发光共振能量转移(BRET)报告等位基因可被CREMEDIATED有条件地打开 重组。这位新开发的记者克服了以前在 动物模型，如要求外部提供激发光和由于 组织吸收，通过使用增强的绿色荧光蛋白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产生迄今已知的最明亮的生物发光信号，并改善了 利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一新的NF2 GEMM。在R33阶段，我们将确定捕食者 GEMM的有效性通过与两种先前被证明诱导抗肿瘤反应的药物进行测试 NF2的可移植模型。这将使我们能够跟踪肿瘤的生长和治疗反应，超过 延长的时间段，在GEMM中密切反映人类疾病。