权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

FOXO signaling and skeletal muscle atrophy

FOXO 信号传导与骨骼肌萎缩

基本信息

批准号：
8183893
负责人：
Andrew Robert Judge
金额：
$ 32.57万
依托单位：
UNIVERSITY OF FLORIDA
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-08-01 至 2016-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8183893
关键词：
Acetylation Acetyltransferase Acquired Immunodeficiency Syndrome Address Aging Area Atrophic Bed rest Binding Boxing CREB-binding protein Cells Chronic Chronic Obstructive Airway Disease DNA Binding Data Deacetylase Deacetylation Denervation Diabetes Mellitus Disease Dominant-Negative Mutation EP300 gene Electrophoretic Mobility Shift Assay Family Gene Targeting Genetic Genetic Transcription Goals HDAC2 gene HDAC4 gene Health Heart failure IGF1 gene Immobilization In Vitro Intervention Lead MLLT7 gene Malignant Neoplasms Measures Mediating Muscle Muscle Fibers Muscular Atrophy Nuclear Nutrient Oligonucleotides Pathway interactions Phosphorylation Physiological Plasmids Post-Translational Protein Processing Protein Acetylation Protein Family Proteins Regulation Reporter Research Role Sepsis Signal Pathway Signal Transduction Signaling Molecule Skeletal Muscle Starvation Stimulus Testing Transcriptional Activation Transfection Work base cell type deprivation human CREBBP protein improved in vivo mRNA Expression muscle form mutant p300/CBP-Associated Factor programs research study response skeletal muscle wasting therapeutic target tool transcription factor wasting

项目摘要

DESCRIPTION (provided by applicant): Skeletal muscle atrophy is a widespread physiological phenomenon and significant health problem associated with muscle disuse (immobilization, bedrest, denervation) and various diseases (cancer, sepsis, AIDS, diabetes, chronic heart failure, chronic obstructive pulmonary disease). However, our understanding of the signaling molecules that regulate muscle mass during an atrophy condition are ill defined. Therefore the long- range goal of our research program is to understand the regulation of signaling pathways that cause muscle atrophy during various conditions. Eventually improved understanding will lead to the identification of the most suitable targets for specific interventions. One family of proteins that is known to cause skeletal muscle atrophy is the FOXO family, and FOXO signaling is increased in a variety of atrophy conditions, including cast immobilization, sepsis, cancer and starvation. However, our understanding of the regulation of FOXO signaling in skeletal muscle is largely confined to the phosphorylation of FOXO factors via the IGF1/PI3K/Akt pathway. While this pathway is clearly important in the regulation of FOXO signaling, another mechanism of regulation, acetylation, appears to be equally important in regulating FOXO signaling, at least in cultured non-muscle cell. However, it is currently unknown whether acetylation of FOXO factors is a regulatory mechanism of FOXO signaling in skeletal muscle during any condition of muscle atrophy. The overall objective of the current proposal is to determine the role of acetyltransferase (HAT) proteins and deacetylase (HDAC) proteins, and acetylation and deacetylation of FOXO factors, in regulating FOXO signaling as it relates to skeletal muscle atrophy. To address this, in Aim 1 we will transfect C2C12 skeletal muscle cells with wild type (WT) or dominant negative (d.n.) HAT proteins and determine their regulation of endogenous FOXO signaling. We will subsequently select one HAT protein for further study in whole skeletal muscle during muscle disuse and cancer. In Aim 2 we will transfect C2C12 skeletal muscle cells with wild type (WT) or dominant negative (d.n.) HDAC proteins and determine their regulation of endogenous FOXO signaling. We will subsequently select one HDAC protein for further study in whole skeletal muscle during muscle disuse and cancer. In Aim 3 we will transfect C2C12 skeletal muscle cells with plasmids encoding FOXO acetylation mutants that mimic either the acetylated or deacetylated forms of FOXO and determine their regulation of FOXO signaling. In each of these Aims we will measure FOXO transcriptional activity, FOXO- DNA binding, FOXO cellular localization, post-translational modifications of FOXO factors, the transcription of a sub-set of known FOXO target genes, and skeletal muscle fiber cross sectional area. The findings from these experiments will lead to a greater understanding of the regulation of FOXO signaling as it relates to skeletal muscle atrophy. PUBLIC HEALTH RELEVANCE: Skeletal muscle wasting is associated with a variety of conditions, including cast immobilization, bed rest, denervation, cancer, sepsis, diabetes and chronic heart failure. In the proposed work we will study a regulatory mechanism of a signaling pathway that is known to regulate skeletal muscle size, to gain a better understanding of how this pathway is controlled. A more comprehensive understanding of the regulation of this pathway is important for the identification of therapeutic targets for muscle wasting.

描述（由申请人提供）：骨骼肌萎缩是一种广泛的生理现象，是与肌肉废用（固定、卧床休息、去神经支配）和各种疾病（癌症、败血症、艾滋病、糖尿病、慢性心力衰竭、慢性阻塞性肺病）相关的严重健康问题。然而，我们对在萎缩状态下调节肌肉质量的信号分子的理解是不明确的。因此，我们研究计划的长期目标是了解在各种条件下导致肌肉萎缩的信号通路的调节。最终，通过加深了解，将确定最适合采取具体干预措施的目标。已知引起骨骼肌萎缩的蛋白质的一个家族是FOXO家族，并且FOXO信号传导在多种萎缩状况中增加，包括管型固定、败血症、癌症和饥饿。然而，我们对骨骼肌中FOXO信号调节的理解主要限于通过IGF1/PI3K/Akt途径磷酸化FOXO因子。虽然该途径在FOXO信号传导的调节中显然是重要的，但另一种调节机制乙酰化在调节FOXO信号传导中似乎同样重要，至少在培养的非肌肉细胞中是如此。然而，目前尚不清楚FOXO因子的乙酰化是否是任何肌肉萎缩条件下骨骼肌中FOXO信号传导的调节机制。目前建议的总体目标是确定乙酰转移酶（HAT）蛋白和脱乙酰酶（HDAC）蛋白的作用，以及FOXO因子的乙酰化和脱乙酰化在调节FOXO信号传导中的作用，因为它与骨骼肌萎缩有关。为了解决这个问题，在目标1中，我们将用野生型（WT）或显性阴性（d. n.）HAT蛋白，并确定其调节内源性FOXO信号。我们随后将选择一种HAT蛋白在肌肉废用和癌症期间的整个骨骼肌中进行进一步研究。在目的2中，我们将用野生型（WT）或显性阴性（d. n.）HDAC蛋白和确定其内源性FOXO信号转导的调节。我们随后将选择一种HDAC蛋白在肌肉废用和癌症期间的整个骨骼肌中进行进一步研究。在目标3中，我们将用编码FOXO乙酰化突变体的质粒转染C2C12骨骼肌细胞，所述突变体模拟FOXO的乙酰化或去乙酰化形式，并确定它们对FOXO信号传导的调节。在这些目标中的每一个中，我们将测量FOXO转录活性、FOXO-DNA结合、FOXO细胞定位、FOXO因子的翻译后修饰、已知FOXO靶基因的子集的转录和骨骼肌纤维横截面积。这些实验的发现将导致对FOXO信号调节的更好理解，因为它与骨骼肌萎缩有关。公共卫生关系：骨骼肌萎缩与多种病症相关，包括石膏固定、卧床休息、去神经支配、癌症、败血症、糖尿病和慢性心力衰竭。在拟议的工作中，我们将研究已知调节骨骼肌大小的信号通路的调节机制，以更好地了解该通路是如何控制的。更全面地了解这一途径的调节对于确定肌肉萎缩的治疗靶点非常重要。