Characterization of Botulinum Neurotoxin A Subtypes

肉毒杆菌神经毒素 A 亚型的表征

• 批准号: 8646866
• 负责人: Eric A. Johnson
• 金额: $ 39.48万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-04-01 至 2017-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8646866
• 关键词: Adverse effects; Area; Base Sequence; Binding; Biologic Characteristic; Biological; Biology; Bontoxilysin; Botulinum Toxin Type A; Botulinum Toxins; Botulism; CCRL2 gene; Categories; Cell model; Cells; Cellular biology; Characteristics; Clinical Treatment; Collaborations; Complex; Data; Development; Diffusion; Disease; Engineering; Future; Genetic Materials; Goals; Human; Immunoprecipitation; In Vitro; Injection of therapeutic agent; Intoxication; Kinetics; Knowledge; Laboratories; Light; Medical; Metalloproteases; Modification; Molecular; Molecular Biology; Mus; Neurons; Outcome; Pharmacologic Substance; Phase; Phenotype; Physiology; Production; Property; Proteins; Recombinants; Research; Research Project Grants; Research Proposals; Sequence Analysis; Serotyping; Site; Solid; Source; Stem cells; Structural Models; Structure; Symptoms; System; Testing; Therapeutic; Therapeutic Uses; Toxic effect; Toxin; Vaccines; Variant; Work; base; botulinum; comparative; improved; in vitro activity; in vivo; insight; novel; protein complex; receptor binding; research study; three-dimensional modeling; toxicity characteristics; trafficking

DESCRIPTION (provided by applicant): Botulinum neurotoxins (BoNTs) are the most poisonous toxins known and are classified as Category A select agents. BoNTs are the causative agent of human botulism, and are used as efficacious pharmaceuticals for treatment of a myriad of neuronal diseases. BoNTs are conventionally subdivided into 7 serotypes (A - G). In recent years, several subtype BoNTs have been identified for most serotypes based on differences in nucleotide sequences. Five subtypes of BoNT/A (A1 - A5) are recognized. Apart from sequence studies and initial data indicating significant functional differences, little researh has been conducted on the molecular and cell biology of the subtype BoNTs. The goal of this research project is to characterize and discern the molecular and cell biology properties of the BoNT/A1-A5 in vitro and in vivo, and relate subtype specific biological characteristics to structural properties. The proposed studies represent a strong partnership among three laboratories with long term and productive collaborations, and the impetus for this application is based upon recent progress in these laboratories on the subtype BoNTs, including that the subtypes differ in molecular characteristics, kinetics and mechanisms of intoxication, and structural features. This proposal will test the overall hypothesis that the five BoNT/A subtypes possess unique in vivo toxicity profiles. Aim 1 will examine the hypothesis that each BoNT/A subtype possesses distinct in vitro toxicity characteristics, and that recombinant BoNT/A subtypes can be produced as source materials for genetic modifications to define functional properties. Aim 2 will test the hypothesis that BoNTs/A1-A5 have distinct cell entry kinetics and in vivo toxicity profiles. Aim 3 will examine the hypothesis that BoNT/A subtypes have unique cell trafficking mechanisms in neuronal cell models, and that the differences in cellular traffickig and biological properties between BoNT/A subtypes are due to specific structural differences of the toxins. The data resulting from this project have the potential to improve clinical treatments with BoNT based pharmaceuticals, as new and modified pharmaceuticals with targeted characteristics and fewer side effects will become necessary. In addition, the characterization of toxin properties will facilitate future developments of targeted anti-BoNT therapeutics that will b effective against all BoNT/A subtypes.

描述（由申请人提供）：肉毒杆菌神经毒素（BONTS）是已知的最有毒的毒素，被归类为类别A选择剂。 BONT是人类肉毒杆菌的病因，被用作有效的药物治疗无数神经元疾病。通常将BONT细分为7种血清型（A -G）。近年来，基于核苷酸序列的差异，已经确定了大多数血清型的几个亚型BONT。识别五个BONT/A（A1 -A5）的亚型。除了序列研究和最初的数据表明功能差异显着，对亚型BONT的分子和细胞生物学进行了很少的研究。该研究项目的目的是表征和辨别BONT/A1-A5在体外和体内的分子和细胞生物学特性，并将亚型特异性生物学特征与结构特性相关联。拟议的研究代表了三个具有长期合作和生产性合作的实验室之间的牢固伙伴关系，并且该应用的动力基于这些实验室在亚型BONTS上的最新进展，包括子类型的分子特征，动力学和动力学和结构特征的分子特征，动力学和机制以及结构特征的不同。该提案将检验总体假设，即五个BONT/A亚型具有独特的体内毒性谱。 AIM 1将检查以下假设：每个BONT/A亚型具有不同的体外毒性特征，并且可以将重组BONT/A亚型作为定义功能特性的遗传修饰的源材料产生。 AIM 2将检验以下假设：BONTS/A1-A5具有独特的细胞进入动力学和体内毒性谱。 AIM 3将检查以下假设：在神经元细胞模型中，BONT/A亚型具有独特的细胞运输机制，并且细胞运输和BONT/A亚型之间的细胞运输和生物学特性的差异是由于毒素的特定结构差异所致。该项目产生的数据有可能通过基于BONT的药物来改善临床治疗，因为具有针对性特征的新和改良的药物将变得更少，副作用将变得更少。此外，毒素特性的表征将促进靶向抗体疗法的未来发展，这些抗BONT疗法将对所有BONT/A亚型有效。