Targeted Delivery of Therapeutics into Motor Neurons for Post-exposure Treatment of Botulism

将治疗药物靶向输送至运动神经元，用于肉毒杆菌中毒的暴露后治疗

基本信息

批准号：
10653914
负责人：
Min Dong
金额：
$ 72.08万
依托单位：
BOSTON CHILDREN'S HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-08-01 至 2026-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10653914
关键词：
Animal Model Animals Antibodies Bacteria Bacterial Toxins Binding Biological Products Bioterrorism Botulinum Toxin Type A Botulinum Toxins Botulism Cardiovascular system Categories Cavia Cessation of life Chimeric Proteins Clostridium Cytosol Dangerousness Data Death Rate Development Disease Disease Outbreaks Drug Delivery Systems Drug Kinetics Engineering Family Family member Half-Life Hour Human Intoxication Lead Medical Membrane Methods Modeling Motor Neurons Mus Muscle Neurons Neurotoxins Paralysed Patients Pharmacodynamics Population Process Production Protein Engineering Proteins Rattus Reproduction spores Residual state Resources Respiratory Failure Respiratory Mechanics Rodent Signal Transduction Specificity Testing Therapeutic Toxic effect Toxin Toxin Conjugates Treatment Efficacy botulinum toxin treatment design efficacy validation exhaust fragment X guinea pig model improved in vivo innovation member motor neuron degeneration nanobodies neutralizing antibody novel pharmacokinetics and pharmacodynamics pre-clinical protein degradation prototype receptor binding success systemic toxicity targeted delivery therapeutic protein tool

项目摘要

Project Summary The family of bacterial toxins, botulinum neurotoxins (BoNTs), produced by spore- forming Gram-positive Clostridium bacteria, are the most potent toxins known. They cause the deadly paralytic disease botulism in humans and animals. These toxins are one of the six most dangerous potential bioterrorism agents (Category A and Tier 1). They target motor neurons with extreme specificity, enter the cytosol of neurons, and block neuronal activity, causing muscle paralysis. A major contributor to the threat of BoNTs is their extremely long half-life within the cytosol of motor neurons – the toxin resides in the cytosol for 4-6 months in humans and causes persistent paralysis for months. To date, no therapeutics can inhibit toxins within the cytosol of neurons. Here we propose to create an effective post-exposure treatment for this top-priority bioterrorism agent and deadly bacterial toxins. This treatment is based on modifying a recently discovered new bacterial toxin BoNT/X and utilizing this engineered protein to deliver a fused neutralizing antibody against BoNTs into the cytosol of motor neurons. Our extensive preliminary studies have provided working prototypes, which completely rescue mice from systemic toxicity of BoNTs in multiple post-exposure models. Here we propose three aims to further validate our hypothesis by (1) refining the chimeric toxin platform; (2) optimizing the nanobody cargo against BoNTs; and (3) establishing pharmacokinetic parameters and validating therapeutic efficacy in both rodent and guinea pig models. Success of these aims will create an effective post-exposure treatment for a top-priority bioterrorism agent, and also develop a novel protein-based delivery platform for targeting motor neurons, thus offering new tools for targeting cytosolic processes and “undruggable” proteins in neurons.

项目概要细菌毒素家族，肉毒杆菌神经毒素 (BoNT)，由孢子产生形成革兰氏阳性梭菌，是已知的最有效的毒素。这些毒素是人类和动物致命的麻痹性疾病肉毒杆菌毒素之一。危险的潜在生物恐怖主义制剂（A 类和 1 级）它们以运动神经元为目标。具有极高的特异性，进入神经元的细胞质，并阻断神经元的活动，导致 BoNT 造成肌肉麻痹的一个主要原因是其极长的半衰期。在运动神经元的细胞质中 – 毒素在人类细胞质中驻留 4-6 个月并导致持续几个月的瘫痪，迄今为止，没有任何治疗方法可以抑制体内的毒素。神经元的细胞质。在此，我们建议针对这一首要任务创建有效的暴露后处理方法这种治疗方法是基于最近修改的一种生物恐怖剂和致命细菌毒素。发现了新的细菌毒素 BoNT/X 并利用这种工程蛋白传递融合蛋白针对 BoNT 的中和抗体进入运动神经元的细胞质。初步研究已经提供了工作原型，可以将小鼠从 BoNT 在多种暴露后模型中的全身毒性在这里我们提出了三个目标。通过（1）完善嵌合毒素平台进一步验证我们的假设；（2）优化针对 BoNT 的纳米抗体货物；以及 (3) 建立药代动力学参数和在啮齿动物和豚鼠模型中验证治疗效果。这些目标的成功将为首要任务创造有效的暴露后治疗生物恐怖主义剂，并开发一种新型的基于蛋白质的靶向电机递送平台神经元，从而为靶向细胞质过程和“不可成药”的蛋白质提供了新的工具神经元。