mTOR-containing Protein Complexes in the Pathogenesis of Tuberous Sclerosis

含 mTOR 的蛋白质复合物在结节性硬化症发病机制中的作用

基本信息

批准号：
7500130
负责人：
Arnold Kristof
金额：
$ 17.72万
依托单位：
MCGILL UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-09-24 至 2012-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7500130
关键词：
Abnormal Cell Address Affinity Affinity Chromatography Apoptosis Apoptosis Regulator Apoptotic Attenuated Benign Binding Biochemical Biogenesis Biological Assay Biopsy Specimen Cell Death Cell Fate Control Cell Nucleus Cell Proliferation Cell model Cell physiology Cells Cessation of life Characteristics Complex Confocal Microscopy Cultured Cells Data Disease Epitopes Equilibrium Event Experimental Models Functional disorder Genetic Transcription Goals Growth Imaging Techniques Immunofluorescence Immunologic Immunohistochemistry In Vitro Inherited Interferon Type II Interferons Kinetics Lesion Life Live Birth Localized Location Lung Lung Transplantation Lung diseases Lymphangioleiomyomatosis Mental Retardation Mitogens Modeling Molecular Molecular Chaperones Multiprotein Complexes Mutation Nature Neurocutaneous Syndromes Normal tissue morphology Nuclear Translocation Nutrient Organ Pathogenesis Patients Population Prevention Principal Investigator Proliferating Protein Biosynthesis Protein Fragment Protein Kinase Proteins Protocols documentation Recombinant Proteins Recombinants Respiratory Failure Ribosomes Role STAT1 gene STAT1 protein Seizures Signal Transduction Sirolimus Skin Smooth Muscle Smooth Muscle Myocytes Standards of Weights and Measures Structure of parenchyma of lung Sturge-Weber Syndrome TSC1 gene TSC2 gene Techniques Therapeutic Therapeutic Agents Tuberous Sclerosis Tuberous sclerosis protein complex Tumor Suppressor Genes Western Blotting base cell growth cell motility human FRAP1 protein human TSC2 protein in vitro Assay inhibitor/antagonist neoplastic novel novel therapeutics pro-apoptotic protein protein kinase C-delta protein protein interaction protein purification research study response scaffold therapeutic target trafficking transcription factor tumor

项目摘要

DESCRIPTION (provided by applicant): Tuberous sclerosis complex (TSC) is a genetically inherited disease of benign tumors in multiple organs. Loss or inactivation of the TSC1 or TSC2 tumor suppressor genes leads to abnormally high activity of the protein mammalian target of rapamycin (mTOR), and excessive cell growth. Arising in one third of patients with tuberous sclerosis complex, lymphangioleiomyomatosis (LAM) features abnormal proliferation of neoplastic smooth muscle-like cells (LAM cells) that leads to cystic destruction of the lung, progressive respiratory failure, and lung transplantation or death. Our long-term goal is to better understand the molecular pathogenesis of tuberous sclerosis complex and LAM. The current therapeutic approach for the treatment of TSC and LAM is to block mTOR activity and cell proliferation. However, mTOR can control a variety of other cellular functions that depend on its presence in distinct macromolecular protein complexes, as well as its cellular location. Our group recently discovered a new mTOR complex that is induced by interferon-(, and contains proteins involved in apoptosis. These include the transcription factor 'signal transducer and activator of transcription-1 (STAT1) and protein kinase C-delta. We propose to investigate the molecular mechanisms that regulate mTOR/STAT1 complex assembly and cellular trafficking, because these events are likely to favor the death of LAM cells, and represent a complementary therapeutic strategy. The specific aims of this proposal are: 1) To determine the biochemical requirements for mTOR/STAT1 protein-protein interactions, 2) To define the nature and requirements for cellular trafficking of the mTOR/ STAT1 complex, 3) To investigate the role of STAT1, and its physical interactions with mTOR, in the control of proliferation and apoptosis in LAM cells, a model of cellular TSC2 deficiency. To examine the molecular requirements for the assembly and cellular trafficking of the mTOR /STAT1 complex, we will employ complementary protein purification techniques, in vitro assays with recombinant proteins, and advanced imaging techniques. Finally, we will investigate the role of mTOR/STAT1 signaling in LAM cell cultures, and biopsy specimens from patients with tuberous sclerosis complex and LAM. Results from these studies will establish a role for rnTOR/STAT1 signaling in the cellular pathophysiology of LAM and tuberous sclerosis, as well as efficient experimental models and platforms to identify novel therapeutic agents.

描述（申请人提供）：结节性硬化症（TSC）是一种多器官良性肿瘤的遗传性疾病。 TSC1 或 TSC2 肿瘤抑制基因的丢失或失活会导致哺乳动物雷帕霉素靶蛋白 (mTOR) 活性异常高，以及细胞过度生长。三分之一的结节性硬化症患者出现淋巴管平滑肌瘤病 (LAM)，其特征是肿瘤性平滑肌样细胞 (LAM 细胞) 异常增殖，导致肺囊性破坏、进行性呼吸衰竭、肺移植或死亡。我们的长期目标是更好地了解结节性硬化症和 LAM 的分子发病机制。目前治疗 TSC 和 LAM 的方法是阻断 mTOR 活性和细胞增殖。然而，mTOR 可以控制多种其他细胞功能，这取决于它在不同大分子蛋白质复合物中的存在及其细胞位置。我们小组最近发现了一种新的 mTOR 复合物，由干扰素诱导，并且含有参与细胞凋亡的蛋白质。其中包括转录因子“信号转导子和转录激活子 1 (STAT1) 和蛋白激酶 C-delta。我们建议研究调节 mTOR/STAT1 复合物组装和细胞运输的分子机制，因为这些事件可能有利于 LAM 细胞的死亡，并且代表了一种补充治疗策略。该提案的具体目标是：1）确定生化要求。为了mTOR/STAT1 蛋白质-蛋白质相互作用，2) 定义 mTOR/STAT1 复合物细胞运输的性质和要求，3) 研究 STAT1 的作用及其与 mTOR 的物理相互作用，在控制细胞增殖和凋亡中的作用LAM 细胞，TSC2 缺陷细胞模型。为了检查 mTOR /STAT1 复合物组装和细胞运输的分子要求，我们将采用互补的蛋白质纯化技术、重组蛋白质的体外测定以及先进的成像技术。最后，我们将研究 mTOR/STAT1 信号在 LAM 细胞培养物以及结节性硬化症和 LAM 患者的活检标本中的作用。这些研究的结果将确定 rnTOR/STAT1 信号在 LAM 和结节性硬化症细胞病理生理学中的作用，以及识别新型治疗药物的有效实验模型和平台。