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.

抽象的肺淋巴管平滑肌瘤病 (LAM) 是一种进展缓慢的疾病，其特征为囊性囊肿肺部遭到破坏，最终导致呼吸衰竭，每百万女性中有 3.4-7.8 人受到影响。从生化角度来说， LAM 是由结节性硬化症复合体 (TSC) 功能丧失引起的，似乎需要激活哺乳动物雷帕霉素复合物靶标 1 (mTORC1) 信号通路的研究。尽管令人鼓舞的临床使用 mTORC1 抑制剂治疗 LAM 的试验结果显示，这些药物受到限制有两个原因：首先，基于雷帕霉素的治疗无法治愈，因为停止治疗后肿瘤会重新生长；第二，长期使用雷帕霉素会产生巨大的副作用，包括免疫紊乱和糖尿病。解决鉴于目前尚未满足的临床需求，该提案将调查三个未解决的机制问题，开发 LAM 长效疗法的长期目标。第一个目标将决定贡献 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，因为无法 mTORC1 靶向疗法诱导毒性仍然是一个尚未解决的障碍。该目标将调查这些细胞中谷氨酰胺代谢的机制以及靶向生长和存活的新方法通过体外和体内药理学和 shRNA 方法治疗 AML 和 LAM 细胞。第二个目标将更好地了解 mTORC1 通路如何调节关键基因的选择性剪接参与 LAM 中看到的“良性转移”表型。虽然 mRNA 剪接受到高度调控并且普遍存在（存在于约 95% 的人类基因中，并且在癌症中经常失调），导致放松管制是未知的。利用我们最近完成的磷酸蛋白质组筛选中的新成果，这个目标将研究 mTORC1 通路如何调节参与“良性肿瘤”的关键基因的剪接最后，第三个目标将确定雌激素如何影响 LAM 转移的关键机制。有助于 LAM 的发病机制，这种疾病几乎只发生在育龄期的女性中。目的是研究雌激素受体信号通过 ERK-Fra1-ZEB 通路的作用。 mTORC1 过度激活，调节 LAM 细胞存活、迁移和侵袭。除了寻址之外 LAM 生物学的基本问题，每个目标都具有临床意义，其中两个目标是动物已提出将细胞生物学发现转化为相关体内模型的研究。综上所述，非常需要对 LAM 的生物学有更深入的了解，并且期望能够成功拟议工作的完成将通过识别新的生物标志物以及影响 LAM 治疗可以特异性消除 TSC 和 TSC 中异常细胞生长、迁移和肿瘤形成的新药林患者。