BRAIN THERAPY USING ULTRASOUND MEDIATED DRUG DELIVERY

大脑疾病，包括癌症和各种形式的 神经退行性疾病，例如阿尔茨海默病和帕金森病 目前，疾病是导致人们患病和死亡的主要原因之一 美国。 这些疾病造成的经济负担是巨大的。 因此，本研究中提出的研究将解决一个关键问题 需要开发安全、非侵入性和非化学方法来增强 通过以下方式将非亲脂性神经药物输送到大脑 使用超声波，已被证明可以增强膜 以非侵入方式传输现象。 但临床使用 超声增强治疗剂在血液中的运输 脑屏障（BBB）尚未得到证实，也没有最佳方案 定义了治疗超声参数。 因此，在本提案中，主要目标是 A) 描绘 用于安全管理超声来调节的参数空间 非亲脂性化合物跨 BBB 的运输，并且 (B) 验证 超声介导转运的安全性和有效性 穿过动物血脑屏障的非亲脂性药物。 为了完成这些 目标，该项目将：（1）开发定制超声装置来执行 细胞培养模型的体外测量范围扩大 BBB 和活体大鼠体内测量，(2) 确定 各种超声波参数（强度、频率、 波形）对极性分子 BBB 渗透性的影响； (3) 确定 不同超声参数对生化指标的影响 BBB细胞的形态特征通过(a)遵循 各种 BBB 特异性酶和膜结合酶的功能表达 蛋白质，(b) 研究形态和细胞学特征 使用标准组织学技术对 BBB 细胞进行分析，以及 (c) 确定 BBB 细胞活力，作为各种超声参数的函数； (4) 确定有效经颅输送的条件 使用模型构建体内大脑模型的超声波； (5)验证 超声介导BBB药物的安全性和有效性 通过检查超声处理的大鼠大脑中的转运 使用标准组织学技术确定病变的存在，并通过 确定经颅超声应用是否可以促进 非亲脂性药物（顺铂）的 BBB 转运可改变平均值 患有一周脑肿瘤的大鼠的存活时间。 按顺序 减少成本和不必要的动物使用，具体目标 2 拟议研究的第 4 部分将使用良好的方法来解决 建立基于生长牛的体外BBB细胞培养模型 细胞培养插管上的脑微血管内皮细胞 (BBMEC)。 预计这项研究还将提供基本见解 超声药物转运的基本机制 BBB 以及设计安全和安全所需的知识 有效的经颅超声传输系统。

Disorders of the brain, including cancer and various forms of neurodegenerative disorders such as Alzheimer's disease and Parkinson's disease are currently among the leading causes of illness and death in the United States. The financial burden of these diseases is enormous. The research proposed in this study will, therefore, address a critical need to develop safe, non-invasive and non-chemical methods to enhance the delivery of nonlipophilic neuropharmaceuticals to the brain through the use of ultrasound, which has been shown to enhance membrane transport phenomena in a non-invasive manner. However, the clinical use of ultrasound-enhanced transport of therapeutic agents across the blood- brain barrier (BBB) has yet to be demonstrated, nor have optimal therapeutic ultrasound parameters been defined. Therefore, in this proposal, the major goals are to A) delineate the parameter space for safe administration of ultrasound to mediate the transport of nonlipophilic compounds across the BBB, and (B) verify the safety and effectiveness of ultrasound in mediating the transport of nonlipophilic drugs across the BBB in animals. To accomplish these goals, the project will: (1) develop custom ultrasound units to perform both an enhanced range of in vitro measurements on a cell-culture model of the BBB and in vivo measurements on live rats, (2) determine the effects of various ultrasonic parameters (intensity, frequency, waveform) on the BBB permeability of a polar molecule; (3) determine the effects of various ultrasonic parameters on biochemical and morphological characteristics of the BBB cells by (a) following the functional expression of various BBB-specific enzymes and membrane-bound protein, (b) studying the morphological and cytological characteristics of BBB cells using standard histological techniques, and (c) determining the BBB cell viability, as a function of various ultrasound parameters; (4) determine the conditions for effective trans-cranial delivery of ultrasound using phantoms constructed to model in vivo brain; (5) verify the safety and effectiveness of ultrasound in mediating BBB drug transport in rats by examining the ultrasound-treated rat brain for the presence of lesions using standard histological techniques, and by determining whether trans-cranial application of ultrasound can promote BBB transport of a nonlipophilic drug (cisplatin) to alter the mean survival times of rats with one-week established brain tumors. In order to reduce costs and the unnecessary use of animals, specific aims 2 through 4 of the proposed study will be addressed using a well- established in vitro BBB cell culture model based on growing bovine brain microvessel endothelial cells (BBMEC) on a cell culture insert. It is anticipated that this study will also provide fundamental insights into the underlying mechanisms of ultrasound drug transport across the BBB as well as the knowledge needed for the design of a safe and effective trans-cranial ultrasound delivery system.