Novel Theranostic Approaches to Identify and Treat Tinnitus Using Functionalized Nanoparticles

使用功能化纳米粒子识别和治疗耳鸣的新治疗诊断方法

• 批准号: 9313977
• 负责人: Magnus Bergkvist
• 金额: $ 13.75万
• 依托单位: SUNY POLYTECHNIC INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-12-01 至 2017-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9313977
• 关键词: Abate; Achievement; Acoustic Stimulation; Address; Adult; Adverse effects; Anatomy; Animal Model; Animals; Antibodies; Area; Attenuated; Auditory; Auditory area; Auditory system; Benchmarking; Binding; Binding Proteins; Biological; Blast Cell; Blood - brain barrier anatomy; Brain; Brain Concussion; Brain region; Capsid; Cells; Cochlear nucleus; Complement; Complex; Contrast Media; Custom; Detection; Development; Diagnosis; Diagnostic; Disabled Persons; Dose; Drug Delivery Systems; Engineering; Evaluation; Exhibits; Exposure to; Fluorescence; Foundations; Frozen Sections; Future; Genes; Goals; Health; Hearing; Histologic; Hyperactive behavior; Image; Imagery; In Vitro; Individual; Inferior Colliculus; Injection of therapeutic agent; LDL-Receptor Related Protein 1; Laboratories; Magnetic Resonance Imaging; Measurement; Microscopy; Molecular; Nanotechnology; Neuraxis; Neurons; Neurosciences; Noise; Peptides; Perception; Peripheral; Pharmaceutical Preparations; Physiological; Proteins; Research; Research Personnel; Resources; Science; Site; Solid; Specificity; Sprague-Dawley Rats; Surface; Symptoms; System; Testing; Therapeutic; Time; Tinnitus; United States; Universities; Weight; Western Blotting; Work; auditory pathway; base; biological research; brain tissue; college; design; differential expression; driving force; falls; fluorophore; in vivo; innovation; interest; nanomaterials; nanoparticle; nanoscale; nanotheranostics; novel; novel strategies; overexpression; receptor binding; relating to nervous system; research facility; targeted delivery; theranostics

DESCRIPTION (provided by applicant): In the United States, about 50 million individuals suffer from tinnitus (perception of sound without overt acoustic stimulation), with circa 3 million people that are disabled and require treatment. Unfortunately, current treatment options for tinnitus are often ineffective and produce inconsistent and sometimes disappointing results. New and innovative approaches for diagnosis and treatment of tinnitus are urgently needed. Here, we introduce a novel concept for tinnitus research and abatement based on the hypothesis that over-expression of genes within brain regions with hyperactive neuronal activity associated with tinnitus can be identified and targeted using multifunctional capsid-based nanoparticles. The effort proposed here leverage a multifunctional capsid-based nanoparticle platform and represents the initial step toward our long-term goal to establish a nanoparticle-based platform to localize and treat tinnitus by attenuating hyperactive neural activity. We will ) evaluate whether nanoparticles can localize to specific targets in vitro and in vivo and ii) optimize the design for ideal localization within the brain and transport across the blood-brain-barrier (BBB) with negligible side effects. Here we seek to evaluate the feasibility of using nanoparticles for in vivo use and establish a solid foundation where future work will enable more in-depth studies using animal models of tinnitus. Specific Aim 1. To establish targeting of specific proteins within the inferior colliculus (IC) using custom-fabricated nanoparticles. Nanoparticles (NPs) will be designed to target proteins demonstrated to be differentially expressed in areas with tinnitus related hyperactivity. Adult Sprague-Dawley rats will be used to evaluate targeting specificity of the NPs. NPs decorated with antibodies that bind receptors in active neurons and loaded with fluorophore will be used for visualization during histological assessment. Two approaches to validate the specificity of NP binding are used: 1. Western blotting will be used to establish that the NPs are capable of binding the protein-of-interest isolated from the IC. This assessment will include, exposure to i) NPs modified with the specific antibody, ii) NPs without the antibody and iii) antibody alone. 2. Microscopy will be used to verify NP binding to the protein of interest in native IC cells. NPs loaded with a fluorescent tag will be localized to quantify differences in NP distribution within and across i) specific subdivisions of the IC ii) other auditory brain regions, as well as, iii) non-auditory brain region. Specific Aim 2. To optimize dose, localization, and transport of MRI detectable custom fabricated nanoparticles across the BBB. Adult Sprague-Dawley rats will be given a systemic injection of custom- designed NPs. They will be modified with a short peptide that facilitates BBB transport (through LPR-1). At multiple time points following injection, nanoparticles with a paramagnetic core and a conjugated fluorescent tag, will be localized: 1. with T1-weighted MRI and 2. cross-validated by fluorescence-based microscopy using frozen sections.

描述（由申请人提供）：在美国，大约5000万个人患有耳鸣（无公开声音刺激的声音感知），大约300万 残障人士需要治疗的人。 不幸的是，耳鸣的当前治疗选择通常无效，并产生不一致的结果，有时令人失望的结果。 迫切需要采用新的和创新的用于诊断和治疗耳鸣的方法。 在这里，我们基于以下假设介绍了一种新颖的耳鸣研究和减排概念，即可以使用基于多功能的capsid纳米粒子来鉴定和靶向具有与耳鸣相关的多动神经元活性的大脑区域内基因的过表达。 这里提出的努力利用了基于CAPSID的多功能纳米颗粒平台，代表了我们朝着长期目标迈出的第一步，以建立一个基于纳米颗粒的平台，通过减弱多动神经活动来定位和治疗耳鸣。 我们将评估纳米颗粒是否可以在体外和体内定位到特定靶标，并且II）优化设计的设计，以实现大脑内部的理想定位以及跨血脑屏障（BBB）的运输，具有可忽略的副作用。 在这里，我们试图评估使用纳米颗粒在体内使用的可行性，并建立一个坚实的基础，在该基础上，未来的工作将使用耳鸣动物模型进行更多的深入研究。 特定目的1。使用自定义的纳米颗粒在下丘（IC）内建立特定蛋白质的靶向。 纳米颗粒（NP）将设计为靶向蛋白质，该蛋白在具有耳鸣相关的多动症的区域差异表达。 成年Sprague-Dawley大鼠将用于评估NP的靶向特异性。 在组织学评估期间，用活性神经元中结合受体并加载荧光团的抗体和富含荧光团的抗体的NP将用于可视化。 使用了两种验证NP结合特异性的方法：1。蛋白质印迹将用于确定NP能够结合与IC分离的利益的蛋白质。 该评估将包括暴露于i）用特异性抗体修饰的NP，ii）无抗体和III）抗体的NP。 2。显微镜将用于验证NP与天然IC细胞中感兴趣的蛋白质的结合。 带有荧光标签的NP 将本地化以量化i）IC II ii）特定细分的NP分布的差异，以及其他听觉大脑区域，以及III）非原告大脑区域。 具体目的2。优化MRI可检测到的定制纳米颗粒在BBB上的剂量，定位和运输。 成年Sprague-Dawley大鼠将被全身注入定制的NP。 它们将通过短肽进行修饰，以促进BBB运输（通过LPR-1）。 在注射后的多个时间点，具有顺磁芯和共轭荧光标签的纳米颗粒将被定位：1。使用冷冻切片通过基于荧光的显微镜进行T1加权MRI和2。交叉验证。