Role of folliculin (FLCN) in lung cell survival

滤泡素 (FLCN) 在肺细胞存活中的作用

基本信息

批准号：
8811465
负责人：
VERA P KRYMSKAYA
金额：
$ 44.92万
依托单位：
UNIVERSITY OF PENNSYLVANIA
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-05-01 至 2017-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8811465
关键词：
5&apos -AMP-activated protein kinase AMP-activated protein kinase kinase Actins Adaptor Signaling Protein Affect Airway Resistance Alveolar Alveolar Cell Anoikis Apoptosis Binding Biochemical Birt-Hogg-Dube Syndrome Bronchoalveolar Lavage Cell Communication Cell Death Cell Polarity Cell Survival Cells Cellular Morphology Complex Cyst Cystic kidney Cytoskeleton Data Disease Distal Down-Regulation Energy Metabolism Epithelial Cells Epithelium Etiology Folliculin Goals Human In Vitro Induced Mutation Kidney Neoplasms Link Lung Lung diseases Maintenance Metabolism Molecular Molecular Target Mus Null Lymphocytes Phosphorylation Pneumothorax Preclinical Testing Production Proteins Publishing Pulmonary Surfactants Respiratory physiology Role Signal Pathway Signal Transduction Small Interfering RNA Testing Transgenic Mice Transgenic Organisms Tuberous sclerosis protein complex Tumor-Suppressor Gene Inactivation alveolar destruction base cell type glucose transport glucose uptake in vivo insight mTOR protein novel novel therapeutic intervention pulmonary function recombinase small hairpin RNA

项目摘要

DESCRIPTION (provided by applicant): Cystic airspace enlargement and spontaneous pneumothorax are major pathological manifestations Birt-Hogg- Dube (BHD) syndrome. Although the etiology of this disease is not known, lung cysts in BHD are linked to autosomal dominant mutational inactivation of tumor suppressor gene folliculin (FLCN). FLCN, a 64-kDa ubiquitously expressed protein with high homology throughout species that lacks apparent functional domain, through adaptor proteins FNIP1/2, binds the 5'-AMP-activated protein kinase (AMPK). Little, however, is known about a role of folliculin (FLCN) in lung cell survival. Our in vitro and in vivo studies demonstrate that FLCN is required for lung cell survival by acting within 5'-AMP-activated protein kinase (AMPK) - tuberous sclerosis complex 2 (TSC2) - mammalian target of rapamycin complex 2 (mTORC2) signaling pathway. To fill a gap in the current understanding of airspace enlargement, we have restricted the scope of this proposal to one testable, central hypothesis that FLCN is required for lung cell survival. To test our hypothesis, in Aim 1, we will determine whether FLCN is required for lung epithelial cell survival in vivo using new transgenic FLCNf/f:SP-C- Cre mice generated in PI's lab. Targeted conditional inducible deletion of FLCN in SP-C-expressing lung epithelial cells will allow us to identify the specific lung epithelial cell type affected by FLCN loss that promotes alveolar space enlargement. In Aim 2 we will examine whether FLCN is required for maintenance of epithelial cell morphology and metabolism. In Aim 3, we will determine FLCN role in regulating activities of Akt and AMPK kinases, and whether pharmacological activation of AMPK will rescue lung epithelial cell survival with targeted inducible deletion of FLCN in FLCNf/f:SP-C-Cre mice. These studies will identify the role of FLCN in lung cell survival, provide insights into the cellular an molecular mechanism of lung epithelial cell-cell contacts, actin cytoskeleton, and metabolism regulated by FLCN. These studies will also establish a mechanistic link between loss of FLCN and cystic airspace enlargement by using our unique novel FLCNf/f:SP- C-Cre transgenic mice and identify potential molecular target(s) for novel therapeutic approaches to treat BHD.

描述（由申请人提供）：囊性空域增大和自发气胸是主要的病理表现，Birt-Hogg- Dube（BHD）综合征。尽管该疾病的病因尚不清楚，但BHD中的肺囊肿与肿瘤抑制基因卵泡蛋白（FLCN）的常染色体显性突变失活有关。 FLCN是一种普遍表达的64 kDa蛋白质，在整个物种中，缺乏明显的功能结构域的高同源性，通过衔接蛋白FNIP1/2结合5'-AMP激活的蛋白激酶（AMPK）。然而，关于卵泡蛋白（FLCN）在肺部细胞存活中的作用知之甚少。我们的体外和体内研究表明，通过在 5' -AMP激活的蛋白激酶（AMPK） - 结节硬化症复合物2（TSC2） - 雷帕霉素复合物2（MTORC2）信号通路的哺乳动物靶标。为了填补当前对空域扩大的理解的空白，我们将该提案的范围限制为一个可检验的，中心假设，即肺部细胞存活需要FLCN。为了检验我们的假设，在AIM 1中，我们将使用PI实验室中产生的新的转基因FLCNF/F：SP-C-CRE小鼠在体内是否需要FLCN。在表达SP-C的肺上皮细胞中FLCN的有针对性的条件诱导缺失将使我们能够确定受FLCN损失影响的特定肺上皮细胞类型，从而促进肺泡空间增大。在AIM 2中，我们将检查是否需要FLCN来维持上皮细胞形态和代谢。在AIM 3中，我们将确定FLCN在调节AKT和AMPK激酶活性中的作用，以及AMPK的药理学激活是否会在FLCNF/F/F：SP-C-C-CRE小鼠中以靶向诱导的FLCN的靶向诱导缺失来挽救肺上皮细胞存活。这些研究将确定FLCN在肺部细胞存活中的作用，为细胞提供肺上皮细胞接触，肌动蛋白细胞骨架和由FLCN调节的代谢的分子机制。这些研究还将通过使用我们独特的新型FLCNF/F：SP-C-CR-CRE转基因小鼠，并确定用于治疗BHD的新型治疗方法，并确定潜在的分子靶标，并确定潜在的分子靶标。