Assay Development for Drugs to Reduce Neuroinflammation

描述（由申请人提供）： 神经退行性疾病如阿尔茨海默病（AD）和多发性硬化症（MS）由于破坏性神经炎症而恶化。 在这些神经退行性疾病中观察到的一个共同主题是正常静止的小胶质细胞被激活成为有效的抗原呈递细胞。 一旦被激活，小胶质细胞就会产生各种细胞因子、趋化因子和神经毒素，从而导致神经炎症。 活化小胶质细胞的机制之一是表达TNF受体超家族成员CD 40。CD 40与其配体CD 154之间的相互作用对于产生免疫应答至关重要。 小胶质细胞的活化还导致许多趋化因子的分泌，包括单核细胞趋化蛋白（MCP-1），其导致新的小胶质细胞J单核细胞募集到MS中的病变部位和AD中的老年斑。最终，增加的CD 40表达加上MCP-1的分泌导致神经炎症的恶化。 靶向特异性抑制或下调CD 40和MCP-1产生的药物的开发将在减少CNS中的神经炎症方面具有很大的效用。 本提案中实验的目标是开发特异性针对CD 40和MCP-1的药物靶标。 这些靶点将用于开发检测方法，以鉴定可用于特异性抑制CD 40和MCP-1表达的药物。 为了实现这一目标，我们将专注于设计和开发靶向特异性抑制MCP-1和CD 40 mRNA的药物，以抑制或下调表达。 在这方面，靶向RNA的小分子试剂，特别是氨基糖苷类抗生素，现在正被积极地用作RNA靶向治疗剂以抑制特定蛋白质的表达。 建议的具体目标如下： 具体目标1：开发靶向MCP-1和CD 40 mRNA的配体。 具体目的2：确定配体抑制MCP-1和CD 40蛋白的表达。一旦我们确定并表征了MCP-1和CD 40 mRNA中的RNA靶点，我们将利用这些信息建立一种高通量筛选试验，用于特异性结合这些RNA的药物;对于这些研究，我们将与NINDS开发的集中式高通量筛选设施合作。可以筛选最佳候选物在体外神经胶质细胞培养系统中下调MCP-1和CD 40表达的能力。

DESCRIPTION (provided by applicant): Neurodegenerative diseases such as Alzheimer's Disease (AD) and Multiple Sclerosis (MS) are exacerbated as a result of damaging neuroinflammation. A common theme observed with these neurodegenerative diseases is the activation of normally quiescent microglia to become potent antigen presenting ceils. Once activated, microglia elaborate a variety of cytokines, chemokines and neurotoxins that contribute to neuroinflammation. One of the mechanisms to activate microglia is the expression of CD40, a member of the TNF receptor superfamily. Interaction between CD40 and its ligand, CD154, is critical for a productive immune response. Activation of microglia also results in secretion of many chemokines, including monocyte chemo attractant protein (MCP-1) that causes the recruitment of new microglia J monocytes to the site of lesions in MS and senile plaques in AD. Ultimately, increased CD40 expression coupled with secretion of MCP-1 results in exacerbation of neuroinflammation. The development of drugs targeted to specifically inhibit or down modulate the production of CD40 and MCP-1 would have great utility in reducing neuroinflammation in the CNS. The goal of the experiments in this proposal is to develop drug targets specific for CD40 and MCP-1. These targets will be used to develop assays to identify drugs that can be utilized to specifically inhibit the expression of CD40 and MCP-1. To accomplish this goal, we will focus on the design and development of drugs targeted to specifically hind to mRNAs of MCP-1 and CD40 as a means to inhibit or down modulate expression. In this regard, small molecule agents that target RNA, notably the aminoglycoside antibiotics, are now actively being pursued as RNA targeted therapeutics to inhibit expression of specific proteins. The following Specific Aims are proposed: Specific Aim 1: To develop ligands that target MCP-1 and CD40 mRNAs. Specific Aim 2: To establish that the ligands inhibit expression of MCP-I and CD40 proteins. Once we have established and characterized the RNA targets in the mRNAs of MCP-1 and CD40, we will use this information to establish a high throughput screening assay for drugs that will specifically bind to these RNAs; for these studies, we will collaborate with the centralized high throughput screening facility that will be developed at NINDS. The best candidates can be screened for their capacity to down modulate MCP-1 and CD40 expression in in vitro glial cell culture systems.