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，大约有 5000 万人患有耳鸣（在没有明显声学刺激的情况下感知声音），其中大约有 300 万人患有耳鸣。 残疾人并需要治疗的人。 不幸的是，目前的耳鸣治疗方案通常无效，并且产生不一致的、有时令人失望的结果。 迫切需要新的创新方法来诊断和治疗耳鸣。 在这里，我们引入了耳鸣研究和减轻的新概念，该概念基于这样的假设：可以使用基于衣壳的多功能纳米颗粒来识别和靶向与耳鸣相关的神经元活动过度活跃的大脑区域内的基因过度表达。 这里提出的工作利用基于衣壳的多功能纳米颗粒平台，代表了我们朝着建立基于纳米颗粒的平台通过减弱过度活跃的神经活动来定位和治疗耳鸣的长期目标迈出的第一步。 我们将）评估纳米颗粒是否可以在体外和体内定位到特定目标，ii）优化设计以实现大脑内的理想定位并穿过血脑屏障（BBB）运输，副作用可以忽略不计。 在这里，我们寻求评估使用纳米颗粒在体内使用的可行性，并为未来的工作使用耳鸣动物模型进行更深入的研究奠定坚实的基础。 具体目标 1. 使用定制纳米颗粒确定下丘 (IC) 内特定蛋白质的靶向。 纳米颗粒（NP）将被设​​计用于靶向在耳鸣相关多动症区域中差异表达的蛋白质。 成年 Sprague-Dawley 大鼠将用于评估 NP 的靶向特异性。 用与活跃神经元中的受体结合的抗体修饰并装载荧光团的纳米粒子将用于在组织学评估期间进行可视化。 使用两种方法来验证 NP 结合的特异性： 1. 将使用蛋白质印迹法来确定 NP 能够结合从 IC 中分离的目标蛋白。 该评估将包括暴露于 i) 用特定抗体修饰的 NP，ii) 没有抗体的 NP，以及 iii) 单独的抗体。 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。 负载荧光标签的纳米粒子 将被本地化以量化 i) IC 的特定细分 ii) 其他听觉脑区域以及 iii) 非听觉脑区域内和之间 NP 分布的差异。 具体目标 2. 优化 MRI 可检测定制纳米粒子在 BBB 上的剂量、定位和传输。 成年 Sprague-Dawley 大鼠将接受定制设计的 NP 的全身注射。 它们将用促进 BBB 运输（通过 LPR-1）的短肽进行修饰。 在注射后的多个时间点，具有顺磁核心和共轭荧光标签的纳米颗粒将被定位：1. 使用 T1 加权 MRI，2. 使用冷冻切片通过基于荧光的显微镜进行交叉验证。