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.

项目摘要这项研究的目的是制造一类RNA适体，以充当AMPA受体抑制剂，并且测试这些适体在小鼠模型中的功效和安全性。这些RNA适体可能成为肌萎缩性侧性硬化症（ALS）治疗的新型药物候选者。 ALS是一个神经退行性疾病的特征是大脑中运动神经元的进行性丧失，脑干和脊髓。目前，ALS药物没有有效地改变疾病的病程。发展更好地控制过度AMPA受体活性的更好抑制剂是发现的有前途的策略新的，更有效的ALS药物。这是因为Ca2+通过异常表达的Ca2+ - 涌入运动神经元中的可渗透AMPA受体导致选择性细胞死亡。 riluzole，两个FDA-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，我们已经确定了一组靶向AMPA受体的14个有效的RNA适体。这些适体更多与常规的小分子抑制剂相比，有效，高度选择性和水溶性。这些特性应使我们能够以最低剂量的剂量使用适合学剂来实现治疗通过更紧密，更有选择地阻止AMPA受体活动在体内的功效，甚至没有剂量依赖性副作用。要检验的假设是将适体用作的组合更好的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鼠标是独特的动物模型在运动神经元中表达异常的Ca2+ - 可渗透AMPA受体亚基，导致最终细胞死亡。因此，AR2小鼠模型是测试我们的AMPA受体适体的理想选择 AMPA受体活性。在AIM 1中，我们建议对14个化学的每一个中的每一个进行大量修改的适体。这些适体将是纯净，稳定且在生物学上的功能性测定。记录。在AIM 2中，我们将在首先测试单独和组合的这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管理，以绕过血液脑屏障。将比较适体治疗和未治疗的小鼠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评估的存活率适体的功效和安全性。这项研究的结果是确定一组适体有效，安全且耐受性良好。如果成功，这项研究应为临床奠定基础对其中一些适体的测试是一种潜在的新ALS疗法。