Retinoid Metabolism in the Adult Heart and Heart Failure

成人心脏和心力衰竭中的类维生素A代谢

• 批准号: 10657290
• 负责人: D Brian Foster
• 金额: $ 75.17万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-01 至 2027-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10657290
• 关键词: Address; Adult; All-Trans-Retinol; Anabolism; Animal Model; Binding; Biological Assay; Cardiac; Cardiac Myocytes; Catabolism; Cellular Retinol Binding Protein; ChIP-seq; Circulation; Clinical; Collaborations; Coronary Arteriosclerosis; Coronary heart disease; Data; Drops; EFRAC; Echocardiography; Elements; Endoplasmic Reticulum; Enzyme Interaction; Enzymes; Epigenetic Process; Etiology; Evaluation; Exhibits; Face; Family member; Fluorescence Microscopy; Functional disorder; Gene Expression; Gene Expression Regulation; Genes; Genetic Transcription; Half-Life; Heart; Heart failure; Homeostasis; Hormones; Human; Human Engineering; Hypertrophic Cardiomyopathy; Image; Impairment; Kinetics; Knock-out; Knockout Mice; Knowledge; Label; Mass Spectrum Analysis; Messenger RNA; Metabolic; Metabolism; MicroRNAs; Mitochondria; Mixed Function Oxygenases; Monitor; Mus; Myocardial Ischemia; Myofibrils; Nuclear; Organ; Outer Mitochondrial Membrane; Oxidoreductase; Patients; Physiological; Post-Translational Protein Processing; Pre-Clinical Model; Primary idiopathic dilated cardiomyopathy; Production; Proteins; Proteomics; Reaction; Relaxation; Reporter; Reporting; Retinaldehyde; Retinoic Acid Receptor; Retinoids; Retinol Binding Proteins; Retinol Metabolism Pathway; Retinol dehydrogenase; Role; Signal Transduction; Site; Specificity; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Techniques; Technology; Testing; Therapeutic; Tissues; Tretinoin; Vitamin A; biosignature; cardiac tissue engineering; cellular engineering; cohort; heart cell; heart function; hemodynamics; human stem cells; in vitro activity; ischemic cardiomyopathy; knock-down; mass spectrometric imaging; medical schools; mouse model; novel; overexpression; pleiotropism; postnatal; preservation; pressure; prevent; programs; promoter; response; transcription factor; transcriptome; translational potential; ultra high resolution

Project Summary This proposal examines the previously unaddressed role of altered retinoid metabolism in heart failure (HF). We have recently shown that in both idiopathic dilated cardiomyopathy (IDCM) and experimental heart failure (HF) there is up to a 40% decline in the cardiac levels of the vitamin A metabolite and potent hormone, all-trans retinoic acid (ATRA), despite adequate vitamin A. We have also shown that direct administration of ATRA prevents HF, in response to pressure-overload. However, implementing ATRA therapy for HF could face headwinds, given the pleiotropy of ATRA signaling and the limited half-life of ATRA in the circulation. Alternatively, therapeutic specificity might best be achieved by targeting the enzymes of cardiac retinoid metabolism. This proposal addresses 3 critical knowledge gaps in our understanding of the mechanism of ATRA metabolism and its impact on the post-natal heart. 1. The enzymes that are responsible for the metabolism of retinol to retinoic acid in the adult mammalian heart are unknown. We have begun to identify the pertinent enzymes in human stem cell-derived cardiomyocytes (hSC-CMs). We show that Dhrs4 regulates the retinaldehyde pool in human stem-cell-derived cardiomyocytes hSC-CMs. We detail a workflow to validate other candidates, leveraging advances hSC-CMs and human engineered heart tissue (hEHT). 2. The pathophysiological mechanisms of a clinically-pertinent cardiac ATRA insufficiency have not been addressed. Knockout of cellular retinol-binding protein 1 (Crbp1) recapitulates the ATRA decline seen in human IDCM. Herein, we show the ATRA decline is sufficient to cause diastolic dysfunction and slow myofibrillar relaxation kinetics. We evaluate the role of the retinaldehyde reductase Dhrs4 by conditional cardiac knockout in the mouse heart. We test the hypothesis that boosting cardiomyocyte ATRA can prevent or even rescue HF and ameliorate cross-bridge cycling. We propose to manipulate DHRS4 and ATRA levels to identify ATRA-sensitive transcriptional programs in mouse hearts, and hEHT. 3. How pervasive the ATRA decline is across HF etiologies is unknown, though proteomic biosignatures of ischemic cardiomyopathy (ICM) and HF with preserved ejection fraction (HFpEF) are consistent with ATRA decline. Even so, another study has shown increased cardiac ATRA in the setting of advanced coronary heart disease. Identifying suitable cohorts for ATRA homeostasis therapy requires that the magnitude and direction of ATRA changes in HF be quantified across HF etiologies. We will quantify retinoids and retinoid-associated protein multiple etiologies including HF with reduced ejection fraction (HFrEF), HFpEF, ICM, and hypertrophic cardiomyopathy (HCM), using state-of-the-art targeted mass spectrometry assays (LC-MS3, isPRM) and examine their distribution within the heart using MALDI-MS Imaging. Restated, this proposal addresses fundamentally novel cardiobiology of a transcriptional master regulator with translational potential, by leveraging the latest advances in human heart cell engineering, novel mouse models, advanced mass spectrometry techniques, and human HF cohorts.

项目摘要 这项建议研究了先前未被提及的维甲酸代谢改变在心力衰竭中的作用。 (Hf)。我们最近发现，在特发性扩张型心肌病(IDCM)和实验性心脏中 衰竭(HF)心脏中维生素A代谢物和强效激素的水平下降高达40%， 全反式维甲酸(ATRA)，尽管有足够的维生素A。我们还表明，直接给药 全反式维甲酸可预防心力衰竭，以应对压力过载。然而，对心力衰竭实施全反式维甲酸治疗可能面临 逆风，鉴于全反式维甲酸信号的多效性和全反式维甲酸在循环中的有限半衰期。或者， 靶向心脏类维A酸代谢的酶可能是实现治疗特异性的最好方法。 这项建议解决了我们在理解全反式维甲酸机制方面的三个关键知识空白 代谢及其对产后心脏的影响。1.负责代谢的酶 视黄醇到视黄酸在成年哺乳动物心脏中的作用尚不清楚。我们已经开始确定相关的 人类干细胞来源的心肌细胞中的酶(HSC-CMS)。我们发现Dhrs4调节 人干细胞来源的心肌细胞中的视黄醛储存池。我们详细介绍了一个工作流来验证其他 利用先进的HSC-CMS和人类工程心脏组织(HEHT)。2. 临床相关的心脏ATRA功能不全的病理生理学机制尚未得到解决。 细胞视黄醇结合蛋白1(Crbp1)的敲除重现了人类IDCM中ATRA的下降。 在此，我们证明ATRA的下降足以导致舒张性功能障碍和缓慢的肌原纤维松弛。 运动学。我们通过条件性心脏基因敲除来评估视黄醛还原酶Dhrs4在小鼠中的作用 心。我们验证了这样的假设，即增强心肌细胞全反式维甲酸可以预防甚至挽救心衰并改善 骑跨桥自行车。我们建议操纵DHRS4和ATRA水平来识别ATRA敏感性 小鼠心脏的转录程序，以及hEHT。3.ATRA下降在心力衰竭病因中的普遍程度 尚不清楚，尽管缺血性心肌病(ICM)和射血保留的心衰的蛋白质组生物学特征 分数(HFpEF)与ATRA下降相一致。尽管如此，另一项研究表明，心脏ATRA增加 在晚期冠心病的背景下。寻找适合全反式维甲酸稳态治疗的队列 要求在不同的心力衰竭病因中量化ATRA在心力衰竭中的变化幅度和方向。我们会 定量检测维甲酸和维甲酸相关蛋白多种病因，包括射血分数降低的心衰 (HFrEF)、HFpEF、ICM和肥厚型心肌病(HCM)，使用最先进的靶向质量 光谱分析(LC-MS3，isPRM)，并使用MALDI-MS成像检查它们在心脏内的分布。 重申一下，这项建议从根本上解决了转录主调控子的新心脏生物学 通过利用人类心脏细胞工程的最新进展，具有翻译潜力的新型小鼠 模型，先进的质谱学技术，和人类高频队列。