ConProject-003

基本信息

批准号：
9981228
负责人：
JOSEPH KISSIL
金额：
$ 11.31万
依托单位：
SCRIPPS FLORIDA
依托单位国家：
美国
项目类别：
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/9981228
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

项目摘要

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.

2 型神经纤维瘤病 (NF2) 是一种显性遗传的常染色体疾病（影响三万分之一），归因于 NF2 基因的杂合性丢失（LOH）。迄今为止最常见的表现该疾病是第八脑神经神经鞘瘤的发展。虽然我们的理解是 NF2 的分子机制在过去 20 年中得到了显着改善，并且许多潜在的治疗靶点已经确定，但该疾病仍缺乏可行的治疗方案。目前的治疗选择是临时的抗症状干预措施，仅限于放射和/或手术并与严重的发病率相关。显然，迫切需要开发治疗方案对于 NF2 患者。目前，开发有效治疗 NF2 的药物的主要障碍之一是缺乏合适的动物模型，以实现候选药物的临床前测试。为了克服这个问题，我们建议开发 NF2 的 GEMM（基因工程小鼠模型），准确反映 NF2 的生物学特性患者神经鞘瘤的发展，纳入转基因报告等位基因，允许颞叶以准确、一致的方式评估候选药物的功效。为了实现这一目标，我们将采用最近描述的 NF2 GEMM，其中两个 Nf2 等位基因在雪旺细胞祖细胞中均失活，通过将 Nf2 条件敲除小鼠与携带 Cre 重组酶等位基因的转基因小鼠进行杂交骨膜素促进剂。然后这些小鼠将与我们开发的报告品系杂交，其中一种新的 Cremediad 可以有条件地开启生物发光共振能量转移 (BRET) 报告基因等位基因重组。这个新开发的报告器克服了以前在动物模型，例如需要外部提供的激发光和由于通过使用增强型 GFP 与增强型荧光素酶变体的融合来实现组织吸收。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，并改善了使用体内光学成像对少量肿瘤细胞进行时空监测。在R21阶段我们将开发并内部验证这个新的 NF2 GEMM。在R33阶段我们将确定预测 GEMM 的有效性，通过使用 2 种先前被证明能引发抗肿瘤反应的药物进行测试 NF2的可移植模型。这将使我们能够在一段时间内跟踪肿瘤的生长和对治疗的反应延长时间段，在密切反映人类疾病的 GEMM 中。