PIMT1 in Red Blood Cell aging in vivo and in vitro

PIMT1在体内和体外红细胞老化中的作用

基本信息

批准号：
10605316
负责人：
Angelo D'Alessandro
金额：
$ 60.58万
依托单位：
UNIVERSITY OF COLORADO DENVER
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-08-01 至 2024-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10605316
关键词：
Acceleration Affect Affinity Age Aging Aldehyde-Lyases American Animal Model Animals Anions Asparagine Aspartate Back Binding Binding Sites Biochemical Biochemical Pathway Biology Blood Blood Banks Blood Transfusion Blood donor Cell Aging Cell Separation Cell Survival Cell membrane Cell physiology Cells Cellular biology Charge Chemicals Childhood Chlorides Data Dehydration Disease Dissection Docking Energy Metabolism Enzymes Erythrocyte Transfusion Erythrocytes Esters Exposure to Failure Flow Cytometry Functional disorder Glucose Glucosephosphate Dehydrogenase Deficiency Glyceraldehyde-3-Phosphate Dehydrogenases Glycolysis Goals Health Hemoglobin Homeostasis Hospitals Human Human body In Vitro Individual Inpatients Knockout Mice Longevity Lung Mediating Medical Membrane Proteins Metabolic Metabolic Pathway Metabolic stress Metabolism Methodology Methylation Methyltransferase Modeling Molecular Mus N-terminal NADP New York Organism Oxidants Oxidation-Reduction Oxidative Stress Oxygen Pathologic Pathology Pathway interactions Pentosephosphate Pathway Peripheral Persons Physiological Play Population Procedures Protein Biosynthesis Protein Methylation Proteins Proteomics Reaction Recycling Regulation Role Sickle Cell Site Succinimides System Testing Time Tissues Toxicology Transfusion Translations Vertebral column age related cell age cell type cofactor deamidation design gene product in vivo metabolomics mouse model mutant novel novel strategies novel therapeutics oxidant stress oxidation oxidative damage repaired senescence sensor stressor tool

项目摘要

ABSTRACT A variety of specific chemical damage occurs as a result of normal cellular senescence, as well as accelerated damage in the context of certain pathologies. One such chemical pathway is the degradation of aspartates into isoaspartyl residues through oxidant damage. As a repair mechanisms, PIMT1 is an enzymatic pathway that methylates isoaspartyl residues, creating an isoaspartyl methyl ester that is capable of then spontaneously reverting into aspartate, thus reversing isoaspartyl damage. Insufficient PIMT1 activity has been associated with increased oxidant stress and shorter cellular and organism lifespan in mice; however, a detailed metabolic and biochemical analysis of the role of PIMT1 has not been elucidated. In this application, we propose to study the role of PIMT1 in cellular aging. While multiple tissues will be analyzed to test general effects of PIMT1, this proposal mainly focusses on a specific central hypothesis regarding effects in red blood cells (RBCs). RBCs are essential to health, and dysfunction of RBCs plays a central role in multiple diseases. In addition, transfusion of RBCs is the single most common inpatient invasive therapy, being given to approximately 1 out of every 70 Americans, annually. RBCs that are transfused are stored (as a logistical necessity) for up to 42 days, during which time they undergo specific cellular and biochemical damage. RBCs are known to lose an essential regulatory function through a key gene product (AE1) in normal cellular aging and in RBC storage. However, the molecular mechanism by which AE1 dysfunction occurs has been unknown. In this application we provide novel data demonstrating that isoaspartyl damage occurs in AE1 of both human and murine RBCs in a domain of AE1 that requires aspartates to function. We likewise present data suggesting that failure of PIMT1 pathways accelerates this damage – however whole animal modeling with deletion of PIMT1 is required to test a mechanistic role. In this context, we offer the following specific aims, designed to critically test hypotheses around the role of PIMT1 mediated repair of oxidant damage. Specific Aim 1: Mechanistic elucidation of the role of protein methylation by PIMT1 in the function and senescence of RBCs. Specific Aim 2: the interaction of increased oxidant stress on PIMT1 and its effects on RBCs aging in vivo and ex vivo (blood storage). PIMT1 null mice will be combined with additional strains designed to isolate metabolic pathways of functional relevance (e.g. G6PD deficient). Advanced experimental methodologies will be applied to these animals in order to isolate cells of particular age and physiological conditions. Finally, the controlled biologies generated from these approaches will be analyzed by cutting edge metabolomic and proteomic methodologies. In aggregate, these studies will advance our understanding of the role of specific pathways of biochemical cellular aging, of the mechanistic role of a conserved repair pathway (PIMT1), and in the context of advanced biochemical analysis and modeling to generate novel mechanistic understanding and critical testing of focused hypotheses.

抽象的由于细胞衰老正常而导致的各种特定的化学损伤发生，并延期在某些病理上的损害。通过氧化剂损伤以及pimt1是一种酶促途径，甲基化的异源居民居民，产生了同甲酯甲基酯，该甲酯能够自发地自发恢复天冬氨酸，从而逆转同甲基损伤。氧化剂应激增加，小鼠的细胞和生物寿命较短）对PIMT1的作用的生化分析尚未在此应用中阐明。 PIMT1在细胞衰老中的作用。提案主要关注有关红细胞（RBC）影响的特定中心假设。对健康至关重要，RBC的功能障碍在多种疾病中起着核心作用。 RBCS是最常见的内科入侵疗法，约有1个以上的70 美国人，每年被输血的RBC（作为后勤上的必要性）长达42天众所周知，在哪个时间经历特定的细胞和生化损害。在正常细胞衰老和RBC存储中，调节功能在正常细胞衰老中的关键基因产物（AE1）。 AE1功能障碍的分子机制在此应用中未知。数据表明，在AE1的域中，在人类和鼠RBC的AE1 AE1中发生异源损伤。这需要Aspartath的功能。加速了这种损害 - 但是，需要使用PIMT1缺失的整个动物建模才能测试机械作用。在这种情况下，我们提供以下特定目标围绕PIMT1介导的氧化剂损伤修复的作用。 PIMT1蛋白甲基化在RBC的功能和衰老中的作用。对PIMT1的氧化应激及其对RBC在体内衰老的影响（血液存储） NULL小鼠将与旨在隔离功能相关的代谢途径的育种菌株结合（例如，G6PD缺乏）。特定年龄和生理条件的细胞。方法将通过尖端代谢组和蛋白质组学方法进行分析研究将建议或理解生化细胞衰老特定途径的核心，保守修复途径（PIMT1）的机械作用，并在先进的生化分析的背景下并建模以产生新的机械理解和对集中假设的批判性测试。