Discovering Biomedically Significant Spider Venom Toxins with Genomics

利用基因组学发现具有生物医学意义的蜘蛛毒毒素

• 批准号: 8100906
• 负责人: Jessica Ellen Garb
• 金额: $ 29.47万
• 依托单位: UNIVERSITY OF MASSACHUSETTS LOWELL
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-04-01 至 2015-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8100906
• 关键词: Address; Animals; Antivenins; Biochemical; Bioinformatics; Cells; Collection; Complex; Coupled; Data; Databases; Development; Evolution; Exhibits; Foundations; Future; Gene Expression; Gene Expression Profile; Gene Family; Genes; Genetic Techniques; Genomics; Gland; Goals; Human; Knowledge; Latrodectus mactans; Lead; Mass Spectrum Analysis; Methods; Molecular; Molecular Probes; Nature; Neurotoxins; Organism; Pain; Pattern; Peptide Mapping; Pharmacologic Substance; Phenotype; Phylogenetic Analysis; Physiological; Physiology; Play; Prize; Production; Property; Proteins; Proteomics; Relative (related person); Resource Development; Resources; Role; Sampling; Scanning; Source; Spider Venoms; Spiders; Techniques; Therapeutic; Tissues; Toxic effect; Toxin; Transcript; Variant; Venoms; Widow; Work; base; black widow spider venom; clinically significant; comparative; improved; innovation; insight; neglect; neurophysiology; neurotoxic; neurotransmission; neurotransmitter release; next generation; novel; public health relevance; research study; tool

DESCRIPTION (provided by applicant): Animal venoms are complex biochemical cocktails capable of diverse and extremely potent physiological effects. Venoms are also highly prized as novel sources of pharmaceuticals and are essential for the development of lifesaving antivenoms. Despite their scientific and clinical significance, the molecular composition and functional activities of many biomedically important venoms remains poorly known. The long-term objective of this proposal is to understand the molecular complexity, function and evolution of black widow spider venom. Black widow spiders possess a highly neurotoxic venom that causes severe pain in humans. While black widow venom has played a critical role in the study of vertebrate neurotransmission, the molecular composition of this venom and its functional and biomedical applications have not been investigated using modern molecular techniques. The proposed work will address this significant knowledge gap through three specific aims. First, a comprehensive black widow venom gland transcriptome will be assembled using next-generation sequencing methods to characterize the genes expressed in venom gland tissue. Second, the set of transcripts involved in venom function will be identified from the transcriptome. This will involve the proteomic technique of peptide fingerprinting to determine the proteins in venom samples using mass spectrometry and bioinformatic screens of a transcriptome database. The next-generation sequencing application of Tag-Profiling will also be used to identify genes that exhibit venom-specific patterns of expression. The third specific aim will examine the functional evolution of black widow venom through cross-species comparisons of venom gene variation that quantify patterns of selection, as well as comparative analyses of gene expression. Phylogenetic analyses of functionally important toxin proteins will also reveal the molecular mechanisms that have contributed to the extreme toxicity of black widow spider venom. When completed, the proposed work will provide an unparalleled view of the molecular diversity, evolution and biomedical utility of black widow venom. The results of this project will also generate significant public genomic resources and provide a foundation for future studies aimed at understanding the molecular mechanisms of toxin functional evolution. PUBLIC HEALTH RELEVANCE: The major goal of this proposal is to determine the molecular composition, functional activities and evolution of black widow spider venom. The project has significant relevance for public health because black widow venom is extremely hazardous to humans, but it is also an essential tool in the study of vertebrate neurophysiology. The proposed work will provide an unprecedented understanding of the molecular basis for the extreme toxicity of black widow spiders and will aid in the development of novel pharmaceuticals and improved antivenoms.

描述(申请人提供)：动物毒液是一种复杂的生化鸡尾酒，能够产生多种和极强的生理作用。毒液作为新的药物来源也备受珍视，对开发救命的抗蛇毒毒素至关重要。尽管它们具有科学和临床意义，但许多生物医学上重要的毒液的分子组成和功能活性仍然鲜为人知。这项建议的长期目标是了解黑寡妇蜘蛛毒液的分子复杂性、功能和进化。黑寡妇蜘蛛拥有一种高度神经毒性的毒液，会给人类带来剧烈的疼痛。虽然黑寡妇毒液在脊椎动物神经传递的研究中发挥了关键作用，但这种毒液的分子组成及其功能和生物医学应用尚未用现代分子技术进行研究。拟议的工作将通过三个具体目标解决这一重大的知识差距。首先，将使用下一代测序方法组装一个全面的黑寡妇毒腺转录组，以表征毒腺组织中表达的基因。其次，参与毒液功能的转录本集合将从转录组中识别出来。这将涉及多肽指纹的蛋白质组学技术，以使用质谱学和转录组数据库的生物信息筛选来确定毒液样本中的蛋白质。标签分析的下一代测序应用程序也将被用于识别表现出毒液特有表达模式的基因。第三个具体目标将通过对毒液基因变异的跨物种比较来量化选择模式，以及对基因表达的比较分析，来研究黑寡妇毒液的功能进化。对具有重要功能的毒素蛋白的系统发育分析也将揭示导致黑寡妇蜘蛛毒液极端毒性的分子机制。完成后，这项拟议的工作将提供一个无与伦比的视角，了解黑寡妇毒液的分子多样性、进化和生物医学用途。该项目的结果还将产生大量的公共基因组资源，并为未来旨在了解毒素功能进化的分子机制的研究提供基础。 公共卫生相关性：这项提案的主要目标是确定黑寡妇蜘蛛毒液的分子组成、功能活性和进化。该项目对公共卫生具有重大意义，因为黑寡妇毒液对人类极其危险，但它也是研究脊椎动物神经生理学的重要工具。这项拟议的工作将对黑寡妇蜘蛛的极端毒性的分子基础提供前所未有的理解，并将有助于开发新的药物和改进的抗蛇毒药物。