Animal testing of RNA aptamers as ALS drug candidate

RNA适体作为ALS候选药物的动物试验

基本信息

批准号：
10646463
负责人：
LI NIU
金额：
$ 40.7万
依托单位：
STATE UNIVERSITY OF NEW YORK AT ALBANY
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-06-15 至 2026-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10646463
关键词：
ALS patients AMPA Receptors Acids Amyotrophic Lateral Sclerosis Animal Model Animal Testing Behavior Biological Biological Assay Blood - brain barrier anatomy Blood capillaries Body Weight Brain Brain Stem Bypass Cell Death Cell Nucleus Central Nervous System Cessation of life Chemicals Circulation Cytoplasm DNA-Binding Proteins Diffuse Disease Dose Down-Regulation Drug or chemical Tissue Distribution Enzymes Epilepsy Evolution FDA approved Failure Functional disorder Glutamates Goals Half-Life Immunohistochemistry In Vitro Individual Infusion procedures Injections Intestinal Absorption Ion Channel Knockout Mice Label Ligands Liquid substance Mediating Metabolic Clearance Rate Modeling Monitor Motor Neurons Mus Nature Nerve Degeneration Neurodegenerative Disorders Neurosciences Research Onset of illness Outcomes Research Pathology Patients Permeability Pharmaceutical Preparations Property Protein Isoforms RNA RNA Editing Research Resistance Rest Ribonucleases Riluzole Safety Site Spinal Cord Stroke Survival Rate Synaptic Transmission Testing Therapeutic Toxic effect Transgenic Mice Treatment Efficacy Water Whole-Cell Recordings Work amyotrophic lateral sclerosis therapy antagonist aptamer blood-brain barrier penetration design drug candidate drug development drug discovery dsRNA adenosine deaminase early detection biomarkers efficacy evaluation efficacy testing in vivo inhibitor lipophilicity motor disorder mouse model nervous system disorder novel prevent protein TDP-43 receptor receptor function research clinical testing response side effect small molecule small molecule inhibitor sporadic amyotrophic lateral sclerosis therapy development water solubility

项目摘要

Project Summary The goal of this research is to make a class of RNA aptamers that act as AMPA receptor inhibitors, and test the efficacy and safety of these aptamers in a mouse model. These RNA aptamers could become a new, potential drug candidate for treatment of amyotrophic lateral sclerosis (ALS). ALS is a neurodegenerative disorder characterized by the progressive loss of motor neurons in the brain, brainstem and spinal cord. Currently none of ALS drugs effectively alters the disease course. Developing better inhibitors to control the excessive AMPA receptor activity is a promising strategy for the discovery of new and more effective ALS drugs. This is because Ca2+ influx through abnormally expressed, Ca2+- permeable AMPA receptors in motor neurons leads to selective cell death. Riluzole, one of the two FDA- approved ALS drugs, actually has anti-glutamatergic properties. Using an in vitro evolution approach, we have identified a group of 14 potent RNA aptamers targeting AMPA receptors. These aptamers are more potent, highly selective and water soluble, as compared with conventional small-molecule inhibitors. These properties should enable us to use aptamers at the lowest dose possible to achieve therapeutic efficacy by more tightly and selectively blocking AMPA receptor activities in vivo with minimal or even no dose-dependent side effects. The hypothesis to be tested is that a combination of using aptamers as better AMPA receptor antagonists and testing them in the AR2 mouse model will allow us to assess the therapeutic efficacy and safety of our aptamers. The AR2 mouse is a unique animal model where an abnormally Ca2+-permeable AMPA receptor subunit is expressed in motor neurons, causing eventual cell death. Thus, the AR2 mouse model is ideal to test our AMPA receptor aptamers in blocking abnormal AMPA receptor activity. In Aim 1, we propose to make a large quantity for each of the 14 chemically modified aptamers. These aptamers will be pure, stable and biologically functional assayed by whole-cell recording. In Aim 2, we will test each of these 14 aptamers, both individually and in combination, in first in AR2 mice. Aptamers will be administrated through intracerebroventricular injection to bypass the blood- brain barrier. Aptamer-treated and -untreated mice will be compared for changes in TDP-43 immunohistochemistry, behaviors, loss of motor neurons and body weight, and survival rate to assess the efficacy and the safety of the aptamers. The outcome of this research is to identify a set of aptamers that are efficacious, safe, and well tolerated. If successful, this research should lay ground work for clinical testing of some of these aptamers as a potential new ALS therapy.

项目概要这项研究的目标是制造一类作为 AMPA 受体抑制剂的 RNA 适体，以及在小鼠模型中测试这些适体的功效和安全性。这些RNA适体可能成为用于治疗肌萎缩侧索硬化症（ALS）的新的潜在候选药物。 ALS 是一种神经退行性疾病，其特征是大脑中运动神经元的逐渐丧失，脑干和脊髓。目前没有任何 ALS 药物能够有效改变病程。发展更好的抑制剂来控制过度的 AMPA 受体活性是这一发现的一个有前途的策略更有效的新型 ALS 药物。这是因为Ca2+通过异常表达流入，Ca2+- 运动神经元中的可渗透性 AMPA 受体导致选择性细胞死亡。利鲁唑（Riluzole），FDA 批准的两种药物之一批准的 ALS 药物实际上具有抗谷氨酸能特性。使用体外进化方法，我们已鉴定出一组 14 个针对 AMPA 受体的有效 RNA 适体。这些适配体更多与传统的小分子抑制剂相比，它具有强效、高选择性和水溶性。这些特性应该使我们能够以尽可能低的剂量使用适体来实现治疗通过更紧密地、选择性地阻断体内 AMPA 受体活性而发挥最小甚至没有作用剂量依赖性副作用。要测试的假设是使用适体作为更好的 AMPA 受体拮抗剂并在 AR2 小鼠模型中测试它们将使我们能够评估我们的适体的治疗功效和安全性。 AR2 小鼠是一种独特的动物模型， Ca2+ 通透性 AMPA 受体亚基异常在运动神经元中表达，最终导致细胞死亡。因此，AR2 小鼠模型非常适合测试我们的 AMPA 受体适体阻断异常的能力。 AMPA 受体活性。在目标 1 中，我们建议对 14 种化学物质中的每一种进行大量生产修饰的适体。这些适体将是纯净的、稳定的并且通过全细胞检测具有生物功能记录。在目标 2 中，我们将首先单独和组合测试这 14 个适体中的每一个。在 AR2 小鼠中。适体将通过脑室内注射来绕过血液- 脑屏障。将比较经过适体处理和未经处理的小鼠 TDP-43 的变化免疫组织化学、行为、运动神经元和体重的损失以及存活率来评估适体的有效性和安全性。这项研究的结果是确定了一组适体有效、安全且耐受性良好。如果成功，这项研究将为临床奠定基础测试其中一些适体作为潜在的新 ALS 疗法。