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.

项目摘要细菌毒素，肉毒神经毒素（BONTS）的家族，由散射产生形成革兰氏阳性细菌是最有效的毒素。他们导致人类和动物的致命麻痹性肉毒杆菌病。这些毒素是最六个危险的潜在生物恐怖剂（A类和第1类）。他们靶向运动神经元具有极高的特异性，输入神经元的细胞质，并阻止神经元活动，从而导致肌肉瘫痪。造成Bonts威胁的主要贡献者是他们的半衰期在运动神经元的细胞质中 - 人类的细胞质中的毒素住所4-6个月并导致数月的持续麻痹。迄今为止，尚无治疗可以抑制毒素神经元的细胞质。在这里，我们建议为这种高优先级创建有效的暴露后治疗生物恐怖剂和致命细菌毒素。这种治疗方法基于修改最近发现了新的细菌毒素BONT/X，并使用该工程蛋白提供融合对抗体的抗体中和运动神经元的细胞质。我们的广泛初步研究提供了工作原型，该原型完全拯救了小鼠在多个暴露后模型中BONT的全身毒性。在这里，我们提出三个目标通过（1）完善嵌合毒素平台进一步验证我们的假设；（2）优化纳米机货物抗管；（3）建立药代动力学参数和验证啮齿动物和豚鼠模型中的治疗功效。这些目标的成功将为高优先级创造有效的暴露后治疗生物恐怖剂，还开发了一种基于蛋白质的新型输送平台，用于靶向电机神经元，因此提供了用于靶向胞质过程的新工具，并且在神经元。