Molecular Coordination of Iron Homeostasis by microRNAs

microRNA 对铁稳态的分子协调

基本信息

批准号：
7939530
负责人：
STEPHEN L CLARKE
金额：
$ 43.61万
依托单位：
OKLAHOMA STATE UNIVERSITY STILLWATER
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-05-01 至 2013-09-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7939530
关键词：
Affect Anemia Animals Applications Grants Cell physiology Chronic Disease Cognitive Computer Simulation Development Diabetes Mellitus Dietary Iron Disease Extramural Activities Foundations Future Gene Expression Gene Expression Regulation Goals Health Homeostasis Impairment In Vitro Individual Iron Iron deficiency anemia Laboratories Lead Light Liver Maintenance Malignant Neoplasms Messenger RNA Metabolism MicroRNAs Modeling Molecular Molecular Profiling Motor Nutrient Physiological Population Positioning Attribute Prevalence Proteins RNA Rattus Research Role Skeletal Muscle Solid Testing Tissues United States Work biological adaptation to stress cell growth regulation disorder prevention experience immune function impaired capacity in vivo innovation insight interdisciplinary approach iron metabolism novel protein expression public health relevance response uptake

项目摘要

DESCRIPTION (provided by applicant): Diseases of iron metabolism continue to be a major health concern in the United States. Iron deficiency remains the most common single nutrient deficiency in the US and individuals experience negative health consequences due to iron deficiency in the absence of anemia, primarily due to alterations in iron metabolism in skeletal muscle. Surprisingly little is known about the molecular mechanisms regulating iron metabolism in skeletal muscle and how alterations in this tissue affects iron homeostasis. The long-term research goal of our laboratory is to advance understanding of how iron metabolism coordinated, and how alterations in iron sensing can lead to the development or prevention of disease. There is evidence suggesting expression of genes involved in iron metabolism is regulated by ID, however, the molecular mechanisms remain poorly characterized. Only recently has the role of small regulatory RNA molecules called microRNAs (miRNAs) been identified as an important mechanism for regulating various cellular processes. Our preliminary in silico analysis of miRNA targets resulted in the identification of mRNAs encoding proteins involved in iron metabolism. Thus our primary objectives are to determine the extent to which expression of miRNAs is regulated in response to ID and to characterize the impact of miRNA expression on potential regulatory targets involved in iron metabolism. We will critically evaluate targets of differentially expressed miRNAs and examine the roles of these targets in cellular iron metabolism. The central hypothesis is that miRNA expression is regulated in response to ID and that changes in expression are associated with changes in the expression target mRNAs resulting in the homeostatic regulation of cellular iron metabolism. To test this hypothesis, this proposal encompasses two specific aims. In Aim 1 we will evaluate and characterize iron-dependent changes in miRNA expression using a weanling rat model of ID. In Aim 2 we will determine the extent to which miRNAs contribute to the regulation cellular iron metabolism through modulating the expression of proteins involved in cellular iron homeostasis. MicroRNA expression profiles in the liver and skeletal muscle will be determined using miRNA microarrays and validated using qPCR. Combining in vivo and in vitro studies, we will identify and characterize target mRNAs and determine the role of these mRNAs in iron metabolism. We will also determine the extent to which changes in miRNA expression are due to alterations in iron status versus alterations in iron sensing. The proposed research is relevant to the maintenance of optimal health in light of the prevalence ID, and by taking an interdisciplinary approach to examine the relationships between iron status and cellular metabolism, we will expand our understanding of how iron contributes to the maintenance of optimal health. PUBLIC HEALTH RELEVANCE: Iron deficiency continues to be the most common nutrient deficiency and is associated with alterations ranging from impairment in immune function, delayed cognitive development, and decreased capacity of work in skeletal muscle. The focus of this project is to examine the extent to which iron status is associated with alterations in microRNA expression that may contribute to some of the tissue-specific effects of iron deficiency. The results of this study will inform our understanding of the molecular mechanisms coordinating the cellular response to iron deficiency and may have implications in understanding alterations in iron metabolism observed in chronic diseases such as cancer and diabetes.

描述（由申请人提供）：在美国，铁代谢疾病仍然是主要健康问题。铁缺乏症仍然是美国最常见的单一营养缺乏症，并且由于缺乏贫血的铁缺乏症而导致的负面健康后果，主要是由于骨骼肌中铁代谢的改变。令人惊讶的是，关于调节骨骼肌铁代谢的分子机制以及该组织中的变化如何影响铁稳态。我们实验室的长期研究目标是提高对铁代谢如何协调的理解，以及铁感应的变化如何导致疾病的发展或预防。有证据表明，与铁代谢有关的基因表达受ID调节，但是，分子机制的表征仍然很差。直到最近，小调节RNA分子称为microRNA（miRNA）的作用被确定为调节各种细胞过程的重要机制。我们对miRNA靶标的计算机分析的初步分析导致鉴定了编码与铁代谢有关的蛋白质的mRNA。因此，我们的主要目标是确定对ID响应的miRNA表达的程度，并表征miRNA表达对铁代谢涉及的潜在调节靶标的影响。我们将批判性地评估差异表达的miRNA的靶标，并检查这些靶标在细胞铁代谢中的作用。中心假设是miRNA表达受到ID的响应调节，并且表达的变化与表达靶标mRNA的变化有关，从而导致细胞铁代谢的稳态调节。为了检验这一假设，该提案涵盖了两个具体目标。在AIM 1中，我们将使用ID的断奶大鼠模型评估和表征miRNA表达中铁的依赖性变化。在AIM 2中，我们将通过调节参与细胞铁稳态的蛋白质的表达来确定miRNA对调节细胞铁代谢的贡献程度。将使用miRNA微阵列确定肝脏和骨骼肌中的microRNA表达谱，并使用qPCR验证。结合体内和体外研究，我们将识别和表征靶标mRNA，并确定这些mRNA在铁代谢中的作用。我们还将确定miRNA表达的变化是由于铁状态与铁感应的改变所致。拟议的研究与鉴于患病率ID的维持与最佳健康有关，并采用跨学科方法来检查铁地位与细胞代谢之间的关系，我们将扩展对铁如何对维持最佳健康的贡献的理解。公共卫生相关性：铁缺乏症仍然是最常见的营养缺乏症，与从免疫功能障碍，认知能力延迟以及骨骼肌工作能力降低的变化有关。该项目的重点是检查铁状态与microRNA表达改变有关的程度，这可能有助于铁缺乏的某些组织特异性作用。这项研究的结果将为我们了解协调细胞对铁缺乏反应的分子机制的理解，并可能对理解在癌症和糖尿病等慢性疾病中观察到的铁代谢的改变具有影响。

项目成果

期刊论文数量（5）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Evaluation of candidate reference genes for quantitative real-time PCR analysis in a male rat model of dietary iron deficiency.

DOI：
10.1186/s12263-021-00698-0
发表时间：
2021-10-02
期刊：
Genes & nutrition
影响因子：
3.5
作者：
Fiddler JL;Clarke SL
通讯作者：
Clarke SL

Comparisons of the iron deficient metabolic response in rats fed either an AIN-76 or AIN-93 based diet.

喂养 AIN-76 或 AIN-93 饮食的大鼠缺铁代谢反应的比较。

DOI：
10.1186/1743-7075-9-95
发表时间：
2012
期刊：
Nutrition & metabolism
影响因子：
4.5
作者：
Davis,McKaleR;Hester,KristenK;Shawron,KristaM;Lucas,EdralinA;Smith,BrendaJ;Clarke,StephenL
通讯作者：
Clarke,StephenL

Influence of microRNA on the maintenance of human iron metabolism.