Aptamer Imaging: A Theranostic Approach to Treat Substance Abuse

适体成像：治疗药物滥用的治疗诊断方法

基本信息

批准号：
8473196
负责人：
Philip K Liu
金额：
$ 32.25万
依托单位：
MASSACHUSETTS GENERAL HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-07-01 至 2014-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8473196
关键词：
AIDS neuropathy Abstinence Acute Affect Affinity Age Amphetamines Anesthesia procedures Anesthetics Animals Antibodies Antisense DNA Astrocytes Award Base Pairing Behavior Binding Biological Assay Biological Markers Biopsy Bipolar Disorder Blood Blood Glucose Brain Brain region Buffers Bypass Caliber Cardiovascular system Catheters Cell Surface Proteins Cells Cerebral Ischemia Cerebrospinal Fluid Cerebrum Chronic Clinical Complex Consensus Sequence Contrast Media Corpus striatum structure Custom Cytokine Receptors DNA Detection Development Diabetes Mellitus Diagnostic Disease Dopamine Antagonists Dose Drug Addiction Electrophoretic Mobility Shift Assay Environment Enzyme-Linked Immunosorbent Assay Enzymes Epigenetic Process Epilepsy Evaluation Exposure to Eye FOS gene Family Fluorescein Fluorescence Frequencies Functional Magnetic Resonance Imaging Future Gender General Anesthesia General Hospitals General Population Generations Genes Genetic Genetic Transcription Genomics Goals HIV-1 Head Head Movements Health Heart Arrest Hour Human Human Activities Image Imaging Techniques Immediate-Early Genes Immunoprecipitation Immunosorbents Implant In Vitro Incubated Individual Indwelling Catheter Infection Infusion procedures Injection of therapeutic agent Intelligence Interleukin-1 Interleukins Intervention Iron Isothiocyanates JUN gene Juice Knock-out Knockout Mice Knowledge Label Laboratories LacZ Genes Learning Length Letters Life Linear Regressions Link Locomotion Lung diseases Macaca mulatta Magnetic Resonance Magnetic Resonance Imaging Maps Massachusetts Measures Mediating Medical Memory Mental Depression Mental Health Mental disorders Messenger RNA Methods Modeling Modification Molecular Molecular Biology Molecular Target Molecular and Cellular Biology Monitor Monkeys Morphologic artifacts Motion Motor Mouse Strains Mus Myocardial Infarction NF-kappa B Needle Sharing Neuroglia Neurologic Neurons Nuclear Extract Nucleic Acids Operative Surgical Procedures Peripheral Pharmaceutical Preparations Plasma Play Population Positron-Emission Tomography Posture Preparation Primates Procedures Process Productivity Promega Promoter Regions Protein Binding Proteins Psyche structure Puncture procedure Quarantine RNA Race Reaction Reagent Recombinants Recovery Reflex action Research Research Personnel Resistance Rest Rewards Rhodamine Risk Rodent Role Running Sampling Scanning Schizophrenia Series Shapes Signal Pathway Signal Transduction Signaling Pathway Gene Site Small Interfering RNA Societies Source Specificity Spinal Cord Spinal Puncture Spottings Substance abuse problem Surface Antigens Symptoms System TNF gene Techniques Technology Testing Therapeutic Time TimeLine Tissues Toxic effect Training Transcription Coactivator Transcription Factor AP-1 Transcription factor genes Transfection Transgenic Mice Transgenic Organisms Translating Translational Research Treatment Efficacy Tumor Necrosis Factor-alpha United States United States National Institutes of Health Urine Variant Vertebral column Visual seizure Weight addiction anticancer treatment aptamer awake base biological adaptation to stress brain cell brain repair clinical application crosslink cytokine design disability dosage driving force drug of abuse ds-DNA experience in vivo indexing innovation inorganic phosphate iron oxide jun D Proteins member molecular imaging mutant nanometer nanoparticle nervous system disorder neuromuscular activity neuropsychiatry neurotoxicity neutravidin nonhuman primate novel nuclease pathogen protein metabolite public health relevance relating to nervous system research study sample fixation severe mental illness theranostics tool transcription factor uptake

项目摘要

DESCRIPTION (provided by applicant): This project will develop and explore Magnetic Resonance (MR) visible probes to image intracellular gene transcription activator proteins. Drug addiction is a major health problem that severely hampers the productivity of many members of our society. While studies indicate that drug addiction results from combined influences of genes and environment, recent studies suggest that epigenetic modifications of gene transcription factors may play an important role in the development of addictions in humans. Advances in cellular and molecular biology in the past century have led to identification of novel gene markers, the signaling pathways they influence and gene activities they regulate. Two such activities that they have shown modification in in studies of drug addiction are intracellular activator protein-1 (AP-1) and nuclear factor kappa- beta (NF-k-b). The inability to produce a functional AP-1 protein to bind at the AP-1 site in a transgenic mouse blocks sensitization of drugs of abuse. With biopsy the only source of tissue for conventional assays, such studies are not permitted in humans. We aim to develop and investigate a targeted MR imaging technique and to apply it for studies of drug addiction in a series of live brains. With a long-term goal of enabling specific manipulation of epigenetic process to non-human primates (NHP), we formulate a hypothesis: short double- stranded (ds) DNA aptamers with the sequence domain for AP-1 protein will function as decoys for endogenous AP-1 protein binding. The aptamers will be linked to an MR-visible contrast agent (superparamagnetic iron oxide nanoparticles-NeutrAvidin, or SPION-NA, 49 nm in diameter), which permits imaging AP-1 proteins in live animal subjects. We have outlined the following steps to validate our hypothesis: Step 1. Design a nuclease-resistant dsAP-1 aptamer with high affinity for AP-1 protein binding Step 2. Quantify rhodamine (Rhd)-labeled dsAP-1 aptamer binding in vitro Step 3. Demonstrate SPION-dsAP1 delivery for MRI and compare in vivo MR signal in C57black6 mice and a mutant strain known to produce no or less AP-1 protein (A-FOS/NSE or FosB knockout mutants) Step 4. Demonstrate Rhd-dsAP1 uptake specificity in cells that express green fluorescence protein (GFP) directed by AP-1 protein [B6;DBA-Tg(Fos-tTA,Fos-EGFP*)1Mmay Tg(tetO-lacZ,tTA*)1Mmay/J] Step 5. Demonstrate dsAP-1 at high dose blocks AP-1 protein-induced activity in mice after amphetamine Step 6. Demonstrate that SPION-dsAP1 delivery allows a window for MRI in live non-human primates and detects elevation of AP-1 protein after amphetamine exposure This application is designed to enable highly innovative and conceptually creative research of molecular targeting of gene products using an unconventional, novel targeted MRI technique for in vivo systems. Because transcription factor binding domains are conserved from rodents to humans, our innovative technique, once validated, has potential for theranostic application, from rodents to primates.

描述（由申请人提供）：该项目将开发和探索磁共振（MR）可见探针，以成像细胞内基因转录激活蛋白。吸毒成瘾是一个重大的健康问题，严重阻碍了我们社会许多成员的生产力。虽然研究表明，药物成瘾是基因和环境的综合影响导致的，但最近的研究表明，基因转录因子的表观遗传修饰可能在人类成瘾的发展中起重要作用。过去一个世纪的细胞和分子生物学进展导致了新型基因标记，它们影响的信号传导途径和调节的基因活性。在药物成瘾研究中，它们在药物成瘾研究中显示的两种活动是细胞内激活蛋白-1（AP-1）和核因子kappa-beta（NF-K-B）。在转基因小鼠中无法产生功能性AP-1蛋白在AP-1位点结合的能力阻止了滥用药物的敏感性。活检是传统测定的唯一组织来源，人类不允许进行此类研究。我们旨在开发和研究有针对性的MR成像技术，并将其应用于一系列活大脑中的药物成瘾研究。通过长期目标，可以对非人类灵长类动物（NHP）进行特定的表观遗传过程的操纵，我们提出了一个假设：具有AP-1蛋白的序列结构域的短双链（DS）DNA适体剂将作为内源性AP-1蛋白结合的诱导型诱导作用。适体将与MR可见对比剂（超帕磁铁氧化铁纳米颗粒 - 中曲霉素或SPION-NA，直径为49 nm）相关，该剂量允许在活动物受试者中成像AP-1蛋白。 We have outlined the following steps to validate our hypothesis: Step 1. Design a nuclease-resistant dsAP-1 aptamer with high affinity for AP-1 protein binding Step 2. Quantify rhodamine (Rhd)-labeled dsAP-1 aptamer binding in vitro Step 3. Demonstrate SPION-dsAP1 delivery for MRI and compare in vivo MR signal in C57black6 mice and a mutant strain known to produce no or less AP-1 protein (A-FOS/NSE or FosB knockout mutants) Step 4. Demonstrate Rhd-dsAP1 uptake specificity in cells that express green fluorescence protein (GFP) directed by AP-1 protein [B6;DBA-Tg(Fos-tTA,Fos-EGFP*)1Mmay Tg(tetO-lacZ,tTA*)1Mmay/J] Step 5. Demonstrate dsAP-1在高剂量阻滞苯丙胺后，AP-1蛋白诱导的小鼠的活性。证明SPION-DSAP1的递送允许在活的非人类灵长类动物中使用MRI窗口，并检测到苯丙胺暴露后的AP-1蛋白的升高，该应用旨在实现高度创新性地对基因构成的构成基因构成的技术，以实现高度创新的基因靶向基因研究。由于转录因子结合结构域是从啮齿动物到人类的保守的，因此我们的创新技术（一旦经过验证）具有从啮齿动物到灵长类动物的疗法应用的潜力。