Molecular and biochemical basis of Lymphangioleiomyomatosis

淋巴管平滑肌瘤病的分子和生化基础

基本信息

批准号：
8612928
负责人：
JOHN BLENIS
金额：
$ 49.81万
依托单位：
HARVARD MEDICAL SCHOOL
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-01-01 至 2014-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8612928
关键词：
Abnormal Cell Address Adverse effects Affect Alternative Splicing Angiomyolipoma Animals Area Benign Biochemical Biochemistry Biogenesis Biological Markers Biological Process Biology Breast Epithelial Cells Cell Proliferation Cell Survival Cells Cellular biology Citric Acid Cycle Clinical Clinical Trials Complex Cytostatics Diabetes Mellitus Disease Disease Progression Energy Supply Equilibrium Estrogen Receptors Estrogens Extracellular Signal Regulated Kinases Family Female Fibronectins Genes Genetic Glucose Glutamine Goals Growth Growth Factor Human Immigration Immune System Diseases Immunologics In Vitro Infiltration Lesion Life Link Lung Lymphangioleiomyomatosis Lymphatic Lymphatic vessel MAPK3 gene Malignant Neoplasms Mediating Messenger RNA Metabolic Metabolism Methods Molecular Mus Mutation Neoplasm Metastasis Normal Cell Null Lymphocytes Nutrient Oncogenic Pathogenesis Pathology Pathway interactions Patients Pharmaceutical Preparations Phenotype Phosphorylation Phosphotransferases Play Pleural effusion disorder Pneumothorax Positioning Attribute Process Property Protein Kinase Proteins Proteomics Publishing RNA Splicing Rattus Receptor Signaling Regulation Research Respiratory Failure Role Serine Signal Pathway Signal Transduction Sirolimus Smooth Muscle Source Spliced Genes Stem cells System TSC1 gene TSC2 gene Testing Therapeutic Therapeutic Intervention Toxic effect Translating Translations Tuberous Sclerosis Tuberous sclerosis protein complex Tumor Suppressor Genes Uterine Fibroids Vascular Endothelial Growth Factors Withholding Treatment Woman Work airway obstruction analog base cell growth cell motility child bearing cytotoxic deprivation drug development drug discovery epithelial to mesenchymal transition expectation human ARMET protein human FRAP1 protein improved in vivo in vivo Model inhibitor/antagonist insight interest loss of function mTOR protein macromolecule meetings migration novel novel strategies protein expression public health relevance pulmonary function response small hairpin RNA transcription factor tumor

项目摘要

Abstract Pulmonary Lymphangioleiomyomatosis (LAM) is a slowly progressing disease characterized by cystic destruction of the lungs and eventual respiratory failure, and affects 3.4-7.8 per million women. Biochemically, LAM is caused by loss of function of the Tuberous Sclerosis Complex (TSC) and appears to require activation of the mammalian target of rapamycin complex 1 (mTORC1) signaling pathway. Despite encouraging clinical trial results from the use of mTORC1 inhibitors to treat LAM, these drugs are limited for two reasons: first, rapamycin-based treatments are not curative as tumors regrow following cessation of treatment; and second, long-term rapamycin usage has tremendous side effects including immune disorders and diabetes. To resolve this currently, unmet clinical need, this proposal will investigate three unresolved mechanistic questions, with the long-term goal of developing long-lasting therapies for LAM. The first aim will determine the contribution of glutamine (Gln) metabolism in AML and LAM cells and its importance in cellular survival as inability of mTORC1-targeted therapies to induce toxicity remains an unaddressed hurdle. This aim will investigate the mechanism of Gln metabolism in these cells as well as new approaches to target the growth and survival of AML and LAM cells via pharmacologic and shRNA methods, both in vitro and in vivo. The second aim will garner a greater understanding of how the mTORC1 pathway regulates the alternative splicing of key genes involved in the "benign metastasis" phenotype seen in LAM. While mRNA splicing is highly regulated and pervasive (occurring in ~95% of human genes and frequently deregulated in cancer), the mechanisms causing deregulation are unknown. Using novel hits from a phospho-proteomic screen we recently completed, this aim will investigate how the mTORC1 pathway regulates the splicing of key genes involved in the "benign metastasis" phenotype of LAM. Finally, the third aim will identify key mechanistic insights into how estrogen contributes to the pathogenesis of LAM, which occurs almost exclusively in females during childbearing years. The aim will investigate the role of estrogen receptor signaling via the ERK-Fra1-ZEB pathway, in the context of mTORC1 hyperactivation, in regulating LAM cell survival, migration and invasion. In addition to addressing basic questions of LAM biology, each of these aims has clinical implications, and in two of the aims animal studies have been proposed to translate the cell biology discoveries into relevant in vivo models. In conclusion, there's a great need for greater understanding of the biology of LAM, and the expectations are that successful completion of the proposed work will impact LAM treatment through identification of new biomarkers as well as novel drugs that can specifically eliminate abnormal cell growth, migration and tumor formation in TSC and LAM patients.

抽象的肺淋巴结肌瘤病（LAM）是一种缓慢发展的疾病，其特征是囊性肺部破坏和最终呼吸道衰竭，每百万妇女影响3.4-7.8。生化， LAM是由结节硬化症复合物（TSC）功能的丧失引起的，似乎需要激活雷帕霉素复合物1（MTORC1）信号通路的哺乳动物靶标。尽管鼓励临床使用MTORC1抑制剂治疗LAM的试验结果，这些药物受到限制，原因有两个：首先，基于雷帕霉素的治疗方法不能治愈，因为肿瘤在停止治疗后再生。第二，长期雷帕霉素的使用具有巨大的副作用，包括免疫疾病和糖尿病。解决目前，这是未满足的临床需求，该建议将研究三个未解决的机械问题，为LAM开发长期疗法的长期目标。第一个目标将决定 AML和LAM细胞中的谷氨酰胺（GLN）代谢及其在细胞存活中的重要性诱导毒性的MTORC1靶向疗法仍然是一个未经应答的障碍。这个目标将调查这些细胞中GLN代谢的机制以及针对生长和存活的新方法 AML和LAM细胞通过药物和shRNA方法在体外和体内。第二个目标 Garner对MTORC1途径如何调节关键基因的替代剪接有更深入的了解参与LAM中看到的“良性转移”表型。而mRNA剪接受到高度调节，并且普遍存在（发生在约95％的人类基因，经常在癌症中引导），导致的机制放松管制未知。使用我们最近完成的磷酸化 - 蛋白质组屏幕的新型命中，这个目的将研究MTORC1途径如何调节与“良性良性”的关键基因的剪接转移”林的表型。最后，第三个目标将确定雌激素如何的关键机理见解有助于LAM的发病机理，该发病机理几乎仅在生育年份发生在女性中。目的将在上下文中研究通过ERK-FRA1-ZEB途径的雌激素受体信号的作用 MTORC1过度活化的，在调节LAM细胞存活，迁移和侵袭中。除了解决 LAM生物学的基本问题，每个目标都具有临床意义，而在两个Aim Anim 已经提出了将细胞生物学发现转化为相关体内模型的研究。综上所述，人们非常需要对LAM的生物学有更深入的了解，而期望是成功的拟议工作的完成将通过识别新的生物标志物以及可以特异性消除TSC和 LAM患者。