Filamin interactions in differentiation, invasion and disease

细丝蛋白在分化、侵袭和疾病中的相互作用

基本信息

批准号：
8437332
负责人：
DAVID A CALDERWOOD
金额：
$ 34.45万
依托单位：
YALE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2003
资助国家：
美国
起止时间：
2003-08-01 至 2017-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8437332
关键词：
Actins Acute Acute Promyelocytic Leukemia Address Animal Model Ankyrin Repeat Binding Binding Sites Biochemical Biological Biological Assay Biological Process Boxing Breast Breast Cancer Cell Bundling Cardiovascular system Cell Adhesion Cell Differentiation process Cells Cleft Palate Collagen Complex Congenital Abnormality Congenital Disorders Congenital Heart Defects Connective Tissue Diseases Crystallography Cytokine Inducible SH2-Containing Protein Cytoskeleton Data Defect Disease Dissection Ehlers-Danlos Syndrome Epithelial Cells Extracellular Matrix Extracellular Matrix Degradation F-Actin FLNC gene Family Gel Genes Genetic Hematopoietic Human Immunoglobulin Domain Integrin Inhibition Integrins Investigation Ligands Lysine Maps Matrix Metalloproteinases Mechanics Mediating Missense Mutation Molecular Muscle Muscle Cells Mutation Myocardium N-terminal Neoplasm Metastasis Neuronal Migration Disorder Neurons Phenotype Point Mutation Process Protein Isoforms Proteins Regulation Resistance Resolution Role Signal Transduction Signaling Protein Site Skeletal Muscle Skeleton Specificity Structure Syndrome Techniques Testing Tretinoin Ubiquitin adhesion receptor base brain malformation cell motility cell type density disease phenotype disease-causing mutation experience fibrosarcoma filamin gain of function human disease interest loss of function malformation malignant breast neoplasm migration mutant periventricular heterotopia protein crosslink public health relevance receptor reconstitution response skeletal skeletal disorder skeletal dysplasia trafficking ubiquitin-protein ligase urinary tract obstruction

项目摘要

DESCRIPTION (provided by applicant): Continued support is requested for our investigation of the roles of filamin in cell differentiation, invasion and disease. Filamins are essential actin crosslinking proteins composed of an N-terminal actin-binding domain followed by 24 immunoglobulin-like domains which interact with numerous cytosolic signaling proteins and transmembrane receptors. Humans have three filamin genes, encoding the widely expressed filamin A and B and largely muscle specific filamin C. Missense point mutations in filamins cause a variety of human diseases, ranging from altered neuronal migration, to cardiac and skeletal muscle defects, and a spectrum of congenital malformations generally characterized by skeletal dysplasias but also including extra-skeletal malformations such as cleft palate, cardiac defects and obstructive uropathy. The actin-binding domain is a hotspot for filamin mutations but, despite dramatic progress in understanding filamin structure and function, how filamin point mutations cause disease remains poorly understood. Furthermore, reduced filamin A expression correlates with increased breast cancer invasion and metastasis, and we recently discovered that loss of filamin increases extracellular matrix (ECM) remodeling and cell invasion. How filamin controls ECM degradation and invasion is unknown. In addition, we have shown that ASB2 (ankyrin repeat-containing protein with a suppressor of cytokine signaling box 2), part of an E3 ubiquitin ligase complex, targets filamins for rapid proteasomal degradation and we suggest that the resultant transient loss of filamin contributes to retinoic acid-induced differentiation of acute promyelocytic leukemia cells. However, the molecular basis for ASB2 function and how loss of filamin influences cell differentiation have not been elaborated. To address the important unanswered questions highlighted above we propose three specific aims which draw on our extensive experience using biochemical, cellular and structural techniques to investigate filamins. Specifically, we will: 1) Characterize the mechanism of ASB2-mediated filamin-degradation and test its role in cell differentiation; 2) Assess the role of filamins in EC remodeling and cell invasion; and 3) Identify cellular phenotypes associated with disease-associated filamin point mutations, revealing potential molecular mechanisms of disease.

描述（由申请人提供）：我们对细丝蛋白在细胞分化、侵袭和疾病中的作用的研究需要持续的支持。细丝蛋白是必需的肌动蛋白交联蛋白由 N 端肌动蛋白结合结构域和 24 个免疫球蛋白样结构域组成，这些结构域与众多胞质信号蛋白和跨膜受体相互作用。人类具有三种细丝蛋白基因，编码广泛表达的细丝蛋白 A 和 B 以及主要是肌肉特异性的细丝蛋白 C。细丝蛋白的错义点突变会导致多种人类疾病，从神经元迁移改变到心肌和骨骼肌缺陷，以及一系列先天性畸形通常以骨骼发育不良为特征，但也包括骨骼外畸形，如腭裂、心脏缺陷和梗阻性尿路病。肌动蛋白结合域是细丝蛋白突变的热点，但是，尽管在了解细丝蛋白结构和功能方面取得了巨大进展，但细丝蛋白点突变如何导致疾病仍然知之甚少。此外，细丝蛋白 A 表达的减少与乳腺癌侵袭和转移的增加相关，我们最近发现细丝蛋白的缺失会增加细胞外基质 (ECM) 重塑和细胞侵袭。细丝蛋白如何控制 ECM 降解和侵袭尚不清楚。此外，我们还发现，ASB2（含有锚蛋白重复序列的蛋白，具有细胞因子信号传导盒 2 的抑制子）是 E3 泛素连接酶复合物的一部分，可靶向细丝蛋白进行快速蛋白酶体降解，我们认为由此产生的细丝蛋白的短暂损失有助于视黄酸诱导急性早幼粒细胞白血病细胞的分化。然而，ASB2 功能的分子基础以及细丝蛋白的缺失如何影响细胞分化尚未得到详细阐述。为了解决上面强调的重要的未解答的问题，我们提出了三个具体目标，这些目标借鉴了我们使用生化、细胞和结构技术研究细丝蛋白的丰富经验。具体来说，我们将： 1）表征ASB2介导的细丝蛋白降解机制并测试其在细胞分化中的作用； 2）评估细丝蛋白在EC重塑和细胞侵袭中的作用； 3) 识别与疾病相关的细丝蛋白点突变相关的细胞表型，揭示疾病的潜在分子机制。