Albumin hitchhiking siRNAs for gene targeting in aged brain

白蛋白搭便车 siRNA 用于老年大脑基因靶向

基本信息

批准号：
10611521
负责人：
Craig Lewis Duvall
金额：
$ 19万
依托单位：
VANDERBILT UNIVERSITY
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：
2022-05-01 至 2025-01-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10611521
关键词：
Abraxane Address Affinity Age Age Months Aging Albumins Alzheimer like pathology Alzheimer&apos s Disease Alzheimer&apos s disease model Alzheimer&apos s disease risk Arthritis Binding Biodistribution Biological Biological Assay Biological Availability Blood Blood - brain barrier anatomy Blood Vessels Blood brain barrier dysfunction Body Weight decreased Brain Bypass C57BL/6 Mouse Cells Cholesterol Circulation Clinical Custom Data Degenerative polyarthritis Deposition Dose Drug Delivery Systems Drug Formulations Drug Kinetics Exclusion Exhibits Fatty Acids Fibrinogen Flow Cytometry Gene Silencing Gene Targeting Genes Genetic Genotype Goals Half-Life Hepatotoxicity Histology Hour Housekeeping Gene Human Hydrophobicity Incidence Individual Inflammatory Intravenous Knee Knee joint Levemir Life Expectancy Link Lipids Lipoprotein Binding Malignant Neoplasms Mediating Messenger RNA Modeling Mus Neurodegenerative Disorders Neurons Organ Outcome Pathologic Penetrance Pharmaceutical Preparations Plasma Positioning Attribute Procedures Property Proteins Publishing Reagent Research Route Scheme Serum Serum Proteins Site Small Interfering RNA Specificity Time Tissues Toxic effect Western Blotting Wild Type Mouse Work aged aging brain apolipoprotein E-3 apolipoprotein E-4 blood-brain barrier disruption brain cell brain tissue cell type cytokine density efficacy study experience genetic risk factor improved in vivo intravenous administration knock-down mouse model nephrotoxicity neurovascular novel novel strategies personalized medicine precision medicine prevent side effect stem treatment strategy tumor uptake

项目摘要

A current hurdle for the treatment of Alzheimer’s disease (AD) is administration of efficacious doses of biological drugs in the brain. The strongest risk factor for AD is aging, which coincides with progressive dysfunction of the blood-brain barrier (BBB) leading to entry and retention of serum proteins that are normally excluded from healthy brain. One of these proteins is albumin, which is not detected in young brain tissue but gradually accumulates in the brain with age. Here, we propose to a novel strategy for treating AD that will leverage albumin as a “natural” carrier to enhance siRNA-mediated gene targeting in the aged brain. We hypothesize that “hitchhiking” siRNA onto albumin will improve siRNA accumulation and gene silencing activity in the aged brain after intravenous administration, thereby providing a customizable strategy to target genes associated with AD. This approach is facilitated by a novel diacyl fatty acid carrier developed in our lab (“EG18”) that can be directly conjugated to siRNA (“siRNA-EG18”). We have relevant preliminary data showing that: 1) siRNA-EG18 has enhanced affinity and specificity for albumin and increased circulation half-life relative to our previously published siRNA-L2 carrier; 2) siRNA-EG18 exerts sustained gene knockdown in a mouse model of osteoarthritis that leads to albumin accumulation in the inflamed knee joint; 3) siRNA-EG18 accumulates in the brains of old but not young mice 24 hours after intravenous delivery of a modest 1 mg/kg dose. Aim 1 of this proposal will build on these results by examining the pharmacokinetics, biodistribution, and toxicity of siRNA-EG18 as a function of age and dosing scheme. Aim 2 will examine the bioactivity of siRNA-EG18 in the aged brain, with a focus on targeting APOE, the strongest known genetic risk factor for AD. Collectively, this proposal will establish working parameters for achieving gene silencing in the aged brain via albumin hitchhiking of siRNA, thereby providing new opportunities for personalized medicine in AD.

目前治疗阿尔茨海默病（AD）的一个障碍是在大脑中施用有效剂量的生物药物。AD 的最强危险因素是衰老，这与导致血脑屏障（BBB）进行性功能障碍相一致。白蛋白是正常情况下被排除在健康大脑之外的血清蛋白的进入和保留，它在年轻的脑组织中检测不到，但随着年龄的增长而逐渐在大脑中积累。这将利用白蛋白作为“天然”我们致力于将siRNA“搭便车”到白蛋白上，以改善静脉注射后老年人大脑中的siRNA积累和基因沉默活性，从而提供针对AD相关基因的可定制策略。我们实验室开发的新型二酰基脂肪酸载体（“EG18”）促进了这种方法，该载体可以直接与 siRNA（“siRNA-EG18”）缀合，我们有相关的初步数据。结果表明：1) 相对于我们之前发表的 siRNA-L2 载体，siRNA-EG18 对白蛋白的亲和力和特异性增强，循环半衰期延长； 2) siRNA-EG18 在骨关节炎小鼠模型中产生持续的基因敲低，导致发炎的膝关节中白蛋白积聚；3) 静脉注射适量 1 后 24 小时，siRNA-EG18 在年老小鼠的大脑中积聚，但年轻小鼠的大脑中则没有。 mg/kg 剂量。该提案的目标 1 将基于这些结果，通过检查 siRNA-EG18 的药代动力学、生物分布和毒性作为函数。目标 2 将检查 siRNA-EG18 在老年大脑中的生物活性，重点是针对已知最强的 AD 遗传风险因素 APOE。总的来说，该提案将建立实现基因沉默的工作参数。通过白蛋白搭 siRNA 的便车来治疗衰老的大脑，从而为 AD 的个性化医疗提供了新的机会。