Imaging Mitochondrial Redox States In Vivo by Hyperpolarized MR

通过超极化 MR 对体内线粒体氧化还原状态进行成像

基本信息

批准号：
8209051
负责人：
LIN Z LI
金额：
$ 40万
依托单位：
UNIVERSITY OF PENNSYLVANIA
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-01-01 至 2015-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8209051
关键词：
2,4-Dinitrophenol Acetoacetates Aging Animal Model Apoptosis Biochemical Biochemical Reaction Biological Assay Biological Markers Biological Process Biomedical Research Biopsy Specimen Breast Cancer Cell Cancer Patient Cancer cell line Carbon Cardiovascular Diseases Cell model Cells Cessation of life Clinical Clinical Treatment Coupled Data Analyses Development Diabetes Mellitus Disease Duchenne muscular dystrophy Electron Transport Equilibrium Esters Event Flavoproteins Future Glucose Hepatic Histology Human Human Biology Hydroxybutyrates Image Indolent Label Link MCF7 cell Magnetic Resonance Magnetic Resonance Imaging Magnetic Resonance Spectroscopy Malignant Neoplasms Mammary Neoplasms Measurement Measures Mediating Metabolic Metabolism Metastatic to Methods Mitochondria Mus NADH Nicotinamide adenine dinucleotide Oxidation-Reduction Oxidoreductase Parkinson Disease Patients Procedures Pyruvate Carboxylase Rare Diseases Rotenone Sequence Analysis Signal Transduction Solutions Succinates System Techniques Time Tissues Translations Validation Xenograft procedure abstracting cancer diagnosis cancer therapy cell growth clinical Diagnosis clinical practice cryogenics disease diagnosis extracellular fluorescence imaging imaging modality in vivo magnetic resonance spectroscopic imaging malignant breast neoplasm melanoma novel public health relevance technique development treatment response tumor

项目摘要

DESCRIPTION (provided by applicant): Abnormalities in mitochondrial metabolism including redox state have been identified under many pathological conditions including cancer, diabetes, aging, and cardiovascular disease. Mitochondrial NAD (nicotinamide adenine dinucleotide)-coupled redox potential NAD+/NADH regulates a number of oxidation-reduction reactions in metabolism, and also mediates key signaling events important for cell growth, survival and apoptosis. There is a great need for a non-invasive method of quantifying and/or measuring mitochondrial NAD+/NADH redox potential in tissues in vivo for the purpose of developing effective approaches for disease diagnosis and treatment. In this project we will develop and validate a clinically translatable and non-invasive hyperpolarized-carbon-13 magnetic resonance spectroscopic imaging (HP-13C-MRSI) method for imaging the mitochondrial redox potential NAD+/NADH in vivo using breast cancer animal models. HP-13C-MR is over 10,000 times more sensitive than conventional MR and has been used to image tissue metabolism. The rationale is to quantify the NAD-coupled redox potential using the equilibrium constant for an enzymatic reaction in mitochondria only, i.e., -hydroxybutyrate + NAD+ ` acetoacetate + NADH + H+, provided that the ratios of acetoacetate/- hydroxybutyrate and the mitochondrial pH can be quantified by MR methods. The proposed method is specific for the NAD+/NADH couple in mitochondria. We will also propose to use 13C-labeled ester probes (e.g. ethyl- hydroxybutyrate) to differentiate between extracellular and intracellular MR signals to obtain the redox potential with higher accuracy. Since our previous studies using cryogenic NADH/flavoprotein fluorescence imaging ex vivo have linked mitochondrial redox state to the metastatic potential of human melanoma and breast cancer in mouse xenografts, this project will also establish a correlation between mitochondrial NAD+/NADH and the metastatic potential of breast cancer, which supports mitochondrial redox potential as a biomarker for cancer diagnosis and treatment response, and a target for therapy. We will investigate whether the mitochondrial redox potential NAD+/NADH measured by the HP-13C-MR method can differentiate an indolent (MCF-7) and a highly metastatic (MDA-MB-231) breast cancer cell line. We will perform redox potential measurements on perfused cell models and mouse xenografts under different mitochondrial metabolic conditions and correlate the results with biochemical assays. We will also validate the techniques on human patients with low or high grade breast tumors to demonstrate its clinical translatability. PUBLIC HEALTH RELEVANCE: Imaging mitochondrial redox states in vivo by hyperpolarized MR Project Summary/Abstract: This proposal is to develop a clinical translatable non-invasive hyperpolarized-carbon-13 magnetic resonance imaging method for quantifying the mitochondrial redox potential NAD+/NADH in vivo in breast tumors. The successful development of the technique will have broad applications in many diseases identified with metabolic abnormalities including cancers, diabetes and cardiovascular diseases. This project may also establish the mitochondrial redox state as a biomarker for cancer diagnosis and treatment response, and a target for therapy.

描述（由申请人提供）：在包括癌症，糖尿病，衰老和心血管疾病在内的许多病理状况下，已经确定了包括氧化还原状态在内的线粒体代谢异常。线粒体NAD（烟酰胺腺嘌呤二核苷酸）耦合的氧化还原电位NAD+/NADH调节代谢中的许多氧化还原反应，还介导了对细胞生长，生存，生存，生存和细胞增多的关键信号事件。非常需要一种非侵入性方法来量化和/或测量体内组织中线粒体NAD+/NADH氧化还原电位，以开发有效的疾病诊断和治疗方法。在该项目中，我们将开发和验证一种可翻译和非侵入性超极化 - 碳13磁共振光谱成像（HP-13C-MRSI）方法，用于对线粒体氧化还原氧化还原电位NAD+/NADH成像，使用乳腺癌动物模型。 HP-13C-MR的灵敏度比常规MR高10,000倍，并且已用于对组织代谢进行图像。理由是使用平衡常数量化NAD偶联的氧化还原电位，仅在线粒体中进行酶促反应，即 - 羟基丁酸 + hydroxybutyrate + NAD +乙酸乙酸乙酸盐 + NADH + NADH + H +，规定乙酸乙酸酯/ - 羟基甲酸/甲酸氢盐的比率是MR MR MR MR MR MR pH PHARDIRAIL。所提出的方法是针对线粒体中NAD+/NADH夫妇的特异性。我们还建议使用13C标记的酯探针（例如乙基羟基丁酸）来区分细胞外和细胞内MR信号，以更高的精度获得氧化还原电位。由于我们先前使用低温NADH/黄素蛋白荧光成像的研究将线粒体氧化还原状态与小鼠异种移植物中人类黑色素瘤和乳腺癌的转移性潜力联系起来，因此该项目还将建立线粒体NAD+/NADH与乳腺癌的乳腺癌的潜在型乳腺癌的乳腺癌的潜能，该项目的相关性反应和治疗的靶标。我们将研究通过HP-13C-MR方法测得的线粒体氧化还原电位NAD+/NADH是否可以区分懒惰（MCF-7）和高度转移（MDA-MB-231）乳腺癌细胞系。我们将在不同的线粒体代谢条件下对灌注细胞模型和小鼠异种移植物进行氧化还原潜在的测量，并将结果与生化测定相关。我们还将验证低或高级乳腺肿瘤的人类患者的技术，以证明其临床转换性。 PUBLIC HEALTH RELEVANCE: Imaging mitochondrial redox states in vivo by hyperpolarized MR Project Summary/Abstract: This proposal is to develop a clinical translatable non-invasive hyperpolarized-carbon-13 magnetic resonance imaging method for quantifying the mitochondrial redox potential NAD+/NADH in vivo in breast tumors.该技术的成功开发将在许多疾病中具有广泛的应用，这些疾病具有代谢异常，包括癌症，糖尿病和心血管疾病。该项目还可以建立线粒体氧化还原状态，作为癌症诊断和治疗反应的生物标志物，并且是治疗的靶标。