Development of De-immunized Botulinum Neurotoxin Type A for Dystonia

用于治疗肌张力障碍的去免疫 A 型肉毒杆菌神经毒素的开发

• 批准号: 7208347
• 负责人: Anne Searls DeGroot
• 金额: $ 15.31万
• 依托单位: EPIVAX, INC.
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-03-01 至 2009-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7208347
• 关键词: Acetylcholine; Acids; Affinity; Algorithms; Alleles; Amino Acids; Antibodies; Antigen Presentation; Beak; Binding; Bioinformatics; Biological; Biological Assay; Blood; Bontoxilysin; Botulinum Toxin Type A; Clinical; Collaborations; Computer Analysis; Computing Methodologies; Detection; Development; Dystonia; Elements; Epitope Mapping; Epitopes; Generations; Genes; Goals; Histocompatibility Antigens Class II; Homology Modeling; Human; Immune response; Immunization; Immunodominant Epitopes; In Vitro; Individual; Injection of therapeutic agent; Intramuscular Injections; Knowledge; Laboratories; Length; Letters; MHC Class II Genes; Mediating; Medical; Modification; Monoclonal Antibodies; Motor Neurons; Movement; Movement Disorders; Mus; Muscle; Mutation; Numbers; Patients; Peptides; Pharmaceutical Preparations; Phase; Process; Production; Public Health; Recombinant Cytokines; Research; Resistance; Site-Directed Mutagenesis; Skeletal Muscle; T-Lymphocyte; T-Lymphocyte Epitopes; TCR Activation; Techniques; Testing; Therapeutic; Toxin; Transgenic Mice; Translations; Treatment Efficacy; Universities; Virus; Work; design; experience; human subject; immunogenic; immunogenicity; improved; in vitro Assay; in vivo; innovation; insight; molecular modeling; neoplastic cell; neutralizing antibody; novel strategies; programs; protein aminoacid sequence; research study; response; stem; therapeutic protein; three dimensional structure; tool; vaccine development

DESCRIPTION (provided by applicant): This revised Exploratory/Developmental (R21) proposal describes innovative research approaches to eliminate the immune response to botulinum neurotoxin type A (BoNT/A) therapy in dystonia patients. The immunogenicity of BoNT/A gives rise to neutralizing antibodies that reduce or eliminate its therapeutic benefits. The proposed research stems from a recognition that development of protein therapeutics in general must be driven by insights into drug-induced immune responses. A generalizable strategy for de- immunization of functional therapeutics will eliminate or at least reduce this major obstacle to the rapid translation of "genes to drugs". This novel approach combines cutting-edge bioinformatics tools with classic immunological assays to create an improved BoNT/A that escapes immunological detection much in the same way as tumor cells and viruses. The target will be the T cell dependent mechanism for antibody generation, which is mediated by peptide-loaded class II MHC-T cell receptor interactions. The primary goals of the project are to use epitope mapping algorithms to computationally identify BoNT/A- derived T cell epitopes. Subsequent verification will be performed experimentally using MHC binding assays and T cell stimulation studies of blood drawn from BoNT/A exposed patients. Immunodominant epitopes will be targeted for modification to reduce their immunogenicity with the aim of preserving the overall fold of the toxin. Computational methods will be used to verify reduced immunogenicity of the modified toxin, and homology modeling will be used to determine the structural effects of these changes. The de-immunized peptide sequences will be assayed for MHC binding and in vitro and in vivo (mouse) T cell stimulation. Public Health Relatedness: Knowledge of the elements of botulinum neurotoxin type A that give rise to its immunogenicity will contribute to the development of improved dystonia therapy.

描述（由申请人提供）：本修订的探索性/发展性（R21）提案描述了消除肌张力障碍患者对A型肉毒杆菌神经毒素（BoNT/A）治疗的免疫反应的创新研究方法。BoNT/A的免疫原性导致产生中和抗体，从而降低或消除其治疗效果。提出的研究源于一种认识，即蛋白质疗法的发展一般必须由对药物诱导的免疫反应的见解驱动。一种可推广的功能疗法去免疫策略将消除或至少减少“基因到药物”快速转化的这一主要障碍。这种新方法将尖端的生物信息学工具与经典的免疫学分析相结合，创造了一种改进的BoNT/A，它可以像肿瘤细胞和病毒一样逃避免疫检测。目标将是T细胞依赖的抗体产生机制，这是由肽负载的II类MHC-T细胞受体相互作用介导的。该项目的主要目标是使用表位映射算法来计算鉴定BoNT/A-衍生的T细胞表位。随后的验证将通过MHC结合分析和T细胞刺激研究从BoNT/A暴露患者的血液中进行实验。免疫显性表位将被靶向修饰，以降低其免疫原性，目的是保留毒素的整体折叠。计算方法将用于验证修饰毒素免疫原性的降低，同源性建模将用于确定这些变化的结构效应。去免疫肽序列将用于MHC结合和体外和体内（小鼠）T细胞刺激。公共卫生相关性：了解A型肉毒杆菌神经毒素中引起其免疫原性的成分将有助于改善肌张力障碍治疗的发展。