Identification of novel pathways causing NF1-driven Schwann cell tumors

鉴定导致 NF1 驱动的雪旺细胞肿瘤的新途径

基本信息

批准号：
10361597
负责人：
NANCY RATNER
金额：
$ 50.06万
依托单位：
CINCINNATI CHILDRENS HOSP MED CTR
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-12-01 至 2026-11-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10361597
关键词：
Affect Agonist Alleles Arginine Binding Biochemical Biochemistry Biotinylation Cancer Model Cell Proliferation Cell Survival Cells Cessation of life Characteristics Complex Data Development Disease Family Fingers Genetic Genetic Diseases Goals Growth Growth Factor Guanosine Triphosphate Phosphohydrolases Heritability Histology Homeostasis Immunohistochemistry In Vitro Individual KRAS2 gene Knockout Mice Location Magnetic Resonance Imaging Mammals Maps Mediating Metabolic Methods Modeling Molecular Biology Techniques Molecular Chaperones Monomeric GTP-Binding Proteins Mus NF1 gene Nervous System Neoplasms Neurofibromatosis 1 Oncogenic Oral Pathway interactions Patients Peripheral Nerves Peroxidases Pharmacology Phosphorylation Phosphorylation Site Preclinical Testing Predisposition Property Prostatic Neoplasms Proteins Reagent Role Schwann Cells Signal Pathway Signal Transduction Survival Analysis Syndrome Tamoxifen Technology Testing Therapeutic Time Tumor Suppressor Proteins Western Blotting base cell type gain of function immunocytochemistry in vivo in vivo evaluation inhibitor interdisciplinary approach loss of function member mouse genetics mouse model mutant neoplastic cell neurofibroma novel paralogous gene prostratin protein complex ras Proteins response self-renewal sensor sermons targeted treatment therapeutic target tumor tumorigenesis

项目摘要

Abstract The NF1 gene product, neurofibromin, was identified in 1990, yet fundamental questions remain unanswered concerning its intracellular location(s), binding partners, and signaling properties. It is known that the large (>200kd) tumor suppressor protein contains a small domain that turns of RAS proteins, so that loss of NF1 function leads to RAS activation on cell stimulation. The remaining NF1 sequence can interact with a chaperone, SPRED1, and other proteins in vitro and/or in specific cell types. In Schwan cells, NF1 loss results in formation of neurofibromas, tumors in the peripheral nerves. It is still unknown which interaction partners and RAS proteins are relevant in Schwann cells. Here we propose to identify NF1 interaction partners, including relevant RAS proteins, that contribute to Schwann cell tumorigenesis. This is because in mammals there are six highly homologous NF1-related RAS paralogs, divided into two families. Our studies rely on interdisciplinary approaches, including cell/mouse genetics, and state of the art biochemical and molecular biology techniques, and our expertise in preclinical testing. Our preliminary data, enabled by new reagents and technologies, supports key roles for canonical RAS proteins in NF1 mutant Schwann cell proliferation. We will now assess the specific roles of RAS proteins in primary Schwann cells in vitro, and in neurofibroma in vivo, and test a potential targeted therapy in vivo. Importantly, whether neurofibromin has Schwann cell-specific interaction partners beyond RAS proteins is unknown. Using proximity biotinylation our preliminary studies identify novel potential NF1-interaction partners in Schwann cells. We will identify complexes containing neurofibromin and these proteins and specific RAS paralogs, and additional complexes that may form on cell stimulation, and test if blocking identified pathways is useful therapeutically.

抽象的 NF1 基因产物神经纤维蛋白于 1990 年被发现，但仍存在一些基本问题关于其细胞内位置、结合伙伴和信号传导仍然没有答案特性。众所周知，大的（>200kd）肿瘤抑制蛋白含有小的转动 RAS 蛋白的结构域，因此 NF1 功能的丧失会导致 RAS 激活细胞刺激。剩余的 NF1 序列可以与伴侣 SPRED1 相互作用，体外和/或特定细胞类型中的其他蛋白质。在雪旺细胞中，NF1 缺失会导致形成神经纤维瘤，周围神经肿瘤。目前尚不清楚是哪一个相互作用伙伴和 RAS 蛋白与雪旺细胞相关。在此我们建议识别 NF1 相互作用伙伴，包括相关 RAS 蛋白，有助于施万细胞肿瘤发生。这是因为在哺乳动物中存在六种高度同源的 NF1 相关基因 RAS旁系同源物，分为两个家族。我们的研究依赖于跨学科方法，包括细胞/小鼠遗传学以及最先进的生物化学和分子生物学技术以及我们在临床前测试方面的专业知识。我们的初步数据由新的支持试剂和技术，支持经典 RAS 蛋白在 NF1 突变体中的关键作用雪旺细胞增殖。我们现在将评估 RAS 蛋白在原发性中的具体作用体外雪旺细胞和体内神经纤维瘤，并测试潜在的靶向治疗体内。重要的是，神经纤维蛋白是否具有雪旺细胞特异性相互作用伙伴 RAS 蛋白之外的情况尚不清楚。使用邻近生物素化我们的初步研究鉴定雪旺细胞中新的潜在 NF1 相互作用伙伴。我们将识别复合物含有神经纤维蛋白和这些蛋白质以及特定的 RAS 旁系同源物，以及其他细胞刺激时可能形成的复合物，并测试阻断已识别的途径是否有用治疗上。