Molecular mechanism of omega-3 response

omega-3反应的分子机制

基本信息

批准号：
8599746
负责人：
JAMES T. BRENNA
金额：
$ 36.92万
依托单位：
CORNELL UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-01-01 至 2017-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8599746
关键词：
11q12 5&apos Untranslated Regions Adult American Anabolism Arachidonic Acids Biochemical Biological Markers Cardiovascular system Catalysis Cells Chemicals Complex DNA Data Degenerative polyarthritis Development Diabetes Mellitus Diet Disease Docosahexaenoic Acids Dominant-Negative Mutation Eicosanoid Production Eicosapentaenoic Acid Euphausiacea Evaluation Fatty Acid Desaturases Fatty Acids Fishes Food Functional RNA Gender Gene Cluster Gene Expression Profile Genes Genetic Genetic Variation Genotype Goals Growth and Development function Haplotypes Health Heart Heterogeneous-Nuclear Ribonucleoproteins Hormonal Hormones Human Human Genetics Individual Industry Infant Intake Intercistronic Region Investigation Knockout Mice Knowledge Lactation Learning Length Link Linolenic Acids Liver MCF7 cell Mediating Mental disorders Metabolic Metabolic Control Metabolic syndrome Molecular Mus Neurologic Nutrient Oils Omega-3 Fatty Acids Open Reading Frames Papio Pathway interactions Phenotype Phosphorylation Polypyrimidine Tract-Binding Protein Polyunsaturated Fatty Acids Positioning Attribute Pregnancy Prevalence Process Protein Isoforms Public Health RNA RNA Interference RNA Splicing Randomized Controlled Trials Regulation Reporting Role Shotgun Sequencing Small Interfering RNA Spliced Genes Supplementation Testing Textbooks Transcript Transcription Initiation Site Untranslated Regions Variant Visual Woman Work base cardiovascular risk factor desaturase dietary supplements fatty acid biosynthesis genome wide association study neglect novel pregnant primary outcome promoter public health relevance response secondary outcome transcriptome sequencing vector

项目摘要

DESCRIPTION (provided by applicant): Omega-3 (?3 or n-3) long chain polyunsaturated fatty acids (LCPUFA, e.g., 22:6n-3, 20:5n-3) are among the most popular dietary supplements used by Americans. Understanding inter-individual response variation requires elucidation of the underlying pathways and the influence of genotypes. LCPUFA biosynthesis is limited by desaturase activity encoded by the fatty acid desaturase gene cluster (FADS1-3, 11q12-13.1). FADS intronic and intergenic SNPs are disproportionately identified as significant in genetic studies (e.g. GWAS). In recent years we showed that all the FADS genes have conserved, highly expressed, and phylogenetically conserved alternative transcripts (AT). A newly described FADS1 AT has desaturase function and new siRNA data show that a specific splicing factor (SF), PTB (hnRNP I) modulates FADS AT abundance. In early 2012, we generated the first Fads3-/- (null) mouse to investigate the as-yet-unknown function of this extensively alternatively spliced gene. AT represent a novel, essential paradigm for omega-3 metabolic control mediated in non-coding regions. Present knowledge of regulatory mechanisms neglecting AT are necessarily incomplete because >95% of human genes produce AT, including the FADS genes. The overall goal is to discover the regulation and function of FADS AT and some of their genetic control via mechanistic investigation. Hypothesis 1: FADS AT are modulated by dietary PUFA and by hormones via splicing factors (SF) in a manner consistent with enhanced catalysis (e.g. FADS1AT1) or inhibition (e.g. FADS3 AT) of desaturation. Hypothesis 2: FADS AT modulation is related to SNPs and haplotypes via splicing factors. Specific Aim 1. Define the transcription start sites (TSS) and UTR of FADS CS and AT. (a) Determine the promoters and full length open reading frames (ORFs) of FADS CS and AT with RNA-Seq using human liver. (b) Determine FADS transcript modulation by ratios of dietary PUFA in mice and by hormones in human cells, thereby establishing AT as intermediate biomarkers for LCPUFA biosynthesis. (c) Develop and implement RNAi of splicing factors and inhibition of their phosphorylation to investigate SF modulation of FADS transcripts, and fatty acid desaturation and composition. Specific Aim 2. Discovery of FADS AT function(s) and modulation. (a) Based on RNA-Seq and RNAi results: (i) Transfect human cells to test for activation or inhibition by specific AT. (ii) Construct vectors for functional studies in human cels, using full length ORF. (b) Characterize the overt and molecular/biochemical phenotype of Fads3 null mice. (c) Investigate the prevalence of human SNPs and haplotypes related to PUFA biosynthesis. Relevance to human health. Understanding of FADS AT regulation, function, and influence on LCPUFA bio- synthesis will enable mechanistic evaluation of interindividual variability in LCPUFA biosynthesis. Human genetic variation and metabolic conditions responsive to omega-3 LCPUFA supplementation will be identified.

描述（由申请人提供）：omega-3（？3或n-3）长链多不饱和脂肪酸（例如LCPUFA，例如22：6n-3，20：5n-3）是美国人使用的最流行的饮食补充剂之一。理解个体间的反应变化需要阐明潜在途径和基因型的影响。 LCPUFA生物合成受到脂肪酸去饱和酶基因簇编码的去饱和酶活性的限制（FADS1-3，11q12-13.1）。 FADS内含子和基因间SNP在遗传研究中被不成比例地识别为重要的（例如GWAS）。近年来，我们表明所有FADS基因都具有保守，高表达和系统发育保守的替代转录本（AT）。新描述的FADS1在HOS Daturase函数上，新的SiRNA数据表明，特定的剪接因子（SF），PTB（HNRNP I）可调节丰富的时尚。在2012年初，我们生成了第一个FADS3 - / - （null）小鼠，以研究该广泛剪接基因的尚未尚未尚未发挥作用。在非编码区域中介导的omega-3代谢控制的新颖，必需的范式。当前对忽略调节机制的知识必然是不完整的，因为> 95％的人类基因在包括时尚基因在内产生的基因。总体目标是通过机械研究发现FADS及其某些遗传控制的调节和功能。假设1：FADS AT由饮食中的PUFA和激素通过剪接因子（SF）调节，其方式与增强的催化（例如FADS1AT1）或抑制（例如FADS3 AT）相一致。假设2：调制时的FADS与SNP和单倍型有关。特定目标1。定义FADS CS和AT的转录起始位点（TSS）和UTR。（a）使用人肝脏确定FADS CS的启动子和全长开放式阅读框（ORF），并使用RNA-Seq使用人肝。（b）确定小鼠饮食PUFA和人类细胞中饮食PUFA比率的FADS转录本调制，从而确定AS为LCPUFA生物合成的AS中间生物标志物。（c）开发和实施剪接因子的RNAi并抑制其磷酸化以研究FADS转录物的SF调节以及脂肪酸的去饱和和组成。特定的目标2。在功能和调制时发现时尚。（a）基于RNA-seq和RNAi结果：（i）转染人类细胞测试特定于AT的激活或抑制。（ii）使用全长ORF构建用于人类CELS功能研究的向量。（b）表征FADS3无效小鼠的公开和分子/生化表型。（c）研究与PUFA生物合成有关的人SNP和单倍型的患病率。与人类健康有关。了解在调节，功能和对LCPUFA生物合成的影响下的时尚将使LCPUFA生物合成中个体变异性的机理评估。将确定对欧米茄-3 lcpufa补充有效的人类遗传变异和代谢状况。