Effect of Heme on mRNA and miRNA Profiles

血红素对 mRNA 和 miRNA 谱的影响

• 批准号: 8432952
• 负责人: HERBERT L BONKOVSKY
• 金额: $ 10.6万
• 依托单位: CAROLINAS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-02-01 至 2014-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8432952
• 关键词: 3' Untranslated Regions; Acute; Acute Intermittent Porphyria; Animal Model; Anti-Inflammatory Agents; Anti-inflammatory; Antioxidants; Aplastic Anemia; Bilirubin; Biliverdine; Binding; Blood; Bone Marrow; Carbon Dioxide; Carbon Monoxide; Catabolism; Cells; Clinical; Complex; Defect; Diamond; Disease; Dose; Down-Regulation; Enzyme Gene; Enzymes; Functional RNA; Gene Expression; Genes; Genetic Transcription; Goals; Health; Heme; Heme Iron; Hemoglobin; Hepatocyte; Homeostasis; Housekeeping; Human; Hydroxymethylbilane Synthase; Inborn Errors of Metabolism; Iron; Kidney; Knowledge; Lead; Leukocytes; Life; Liver; Mediating; Messenger RNA; Metabolic Pathway; Metabolism; Metalloporphyrins; MicroRNAs; Mitochondria; Modeling; Mus; Mutation; Nuclear Receptors; Organ; Oxygen; Pathway interactions; Patients; Peptide Hydrolases; Peripheral Blood Mononuclear Cell; Personal Satisfaction; Physiological; Play; Porphyrias; Porphyrins; Prevention; Principal Investigator; Property; Proteins; Protoporphyrins; RNA; Reaction; Regulation; Reporting; Role; Seeds; Seminal; Signal Pathway; Signaling Molecule; Spleen; Syndrome; Tissues; Toxic effect; Translations; Up-Regulation; base; blood forming organ; comparative; enzyme activity; heme oxygenase-1; improved; in vitro Model; in vivo; insight; intravenous administration; mRNA Stability; novel; programs; public health relevance; synthetic enzyme; uptake

DESCRIPTION (provided by applicant): Heme [iron protoporphyrin] is a primordial macrocycle upon which nearly all life on earth depends. It has manifold known functions and diverse properties. The importance of a normal pathway and regulation of heme metabolism is underscored by the seriousness of diseases in which there are defects in heme homeostasis. For example, the porphyrias are a group of diseases in which there are defects in normal heme synthesis, due mainly to inborn errors of metabolism that produce deficient activities of the enzymes of normal porphyrin and heme synthesis. Genes and their products of particular importance in heme metabolism are ALA synthase 1 [ALAS1] and heme oxygenase 1 [HMOX1], respectively, the rate-controlling enzymes of heme synthesis and catabolism. Hydroxymethylbilane synthase [HMBS], the third enzyme of the heme biosynthetic pathway, is deficient in acute intermittent porphyria, the most severe of the acute porphyrias. Recently, we reported important novel effects of miRNAs-122, -196, and -let 7 on expression of HMOX1 and its key repressor BACH1, and we recently discovered new and heretofore unexpected roles of proteasomal and other protease pathways that regulate levels of ALAS1, BACH1, and HMOX1. We also have discovered changes caused by heme excess vs heme deficiency on reciprocal expression of miRNAs and mRNAs in human hepatocytes. Heme, which is the treatment of choice for acute porphyric attacks and for their prevention, may have deleterious effects on the liver and other tissues and organs, especially with repeated use. Our recent microarray results in human Huh-7 cells have unveiled novel effects of heme excess vs heme deficiency on several genes and pathways. It is important now to assess the potential physiological importance of alterations in miRNA and mRNA profiles in in vitro models and whether and to what extent similar changes occur in vivo, in animal models of AIP and in humans with acute porphyrias. We hypothesize that heme and other metalloporphyrins regulate ALAS1, HMBS and HMOX1 expression by altering selected microRNAs and mRNA profiles Our specific aims are as follows: Aim #1: To delineate effects and mechanisms whereby selected microRNAs modulate expression of ALAS1, HMBS, and HMOX1 and/or their key regulators in human hepatocytes. Aim #2 To characterize the effects of selected, physiologically relevant doses of heme on the livers, kidneys, spleens, and bone marrows of mice with mutations that resemble those seen in human AIP and on peripheral blood mononuclear cells of patients with AIP who are receiving heme as therapy. Results of these studies will provide important new insights into the manifold and critical functions of heme in health and disease, and in our understanding at both the basic and clinical level.

描述（由申请人提供）：血红素[铁原卟啉]是一种原始大环化合物，地球上几乎所有生命都依赖于它。它具有多种已知的功能和多种特性。血红素稳态存在缺陷的疾病的严重性强调了血红素代谢正常途径和调节的重要性。例如，卟啉症是一组正常血红素合成存在缺陷的疾病，主要是由于先天性代谢错误导致正常卟啉和血红素合成的酶活性缺陷。在血红素代谢中特别重要的基因及其产物分别是 ALA 合成酶 1 [ALAS1] 和血红素加氧酶 1 [HMOX1]，分别是血红素合成和分解代谢的速率控制酶。羟甲基胆烷合酶 [HMBS] 是血红素生物合成途径的第三种酶，在急性间歇性卟啉症（最严重的急性卟啉症）中存在缺陷。最近，我们报道了 miRNA-122、-196 和 -let 7 对 HMOX1 及其关键阻遏物 BACH1 表达的重要新作用，并且我们最近发现了调节 ALAS1、BACH1 和 HMOX1 水平的蛋白酶体和其他蛋白酶途径的新的和迄今为止意想不到的作用。我们还发现血红素过量与血红素缺乏引起的人肝细胞中 miRNA 和 mRNA 相互表达的变化。血红素是急性卟啉症发作和预防的首选治疗方法，但可能对肝脏和其他组织和器官产生有害影响，特别是重复使用时。我们最近在人类 Huh-7 细胞中的微阵列结果揭示了血红素过量与血红素缺乏对多个基因和途径的新影响。现在重要的是评估体外模型中 miRNA 和 mRNA 谱变化的潜在生理重要性，以及体内、AIP 动物模型和患有急性卟啉症的人类是否以及在多大程度上发生类似的变化。我们假设血红素和其他金属卟啉通过改变选定的 microRNA 和 mRNA 图谱来调节 ALAS1、HMBS 和 HMOX1 表达。我们的具体目标如下：目标 #1：描绘选定的 microRNA 调节人肝细胞中 ALAS1、HMBS 和 HMOX1 和/或其关键调节因子表达的作用和机制。目标#2 表征选定的生理相关剂量的血红素对具有与人类 AIP 相似突变的小鼠的肝脏、肾脏、脾脏和骨髓的影响，以及对接受血红素治疗的 AIP 患者外周血单核细胞的影响。这些研究的结果将为血红素在健康和疾病中的多种关键功能以及我们在基础和临床层面的理解提供重要的新见解。