Comparative venomes of recluse spiders and kind: treatments and toxin discovery

隐士蜘蛛及其种类的毒液比较：治疗方法和毒素发现

基本信息

批准号：
8879895
负责人：
Greta Jeanne Binford
金额：
$ 26.6万
依托单位：
LEWIS AND CLARK COLLEGE
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-07-01 至 2019-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8879895
关键词：
Amino Acids Animals Antivenins Basic Science Bite Chemicals Complementary DNA Complex Complex Mixtures Computational Biology Cytotoxin Data Databases Development Diagnostic Drug Targeting Effectiveness Evolution Family Freezing Funding Gene Expression Profile Gene Family Genealogical Tree Genome Gland Goals Health Homologous Gene Human Individual Insecticides Lesion Life Medical Methods Modeling Molecular Evolution Morphologic artifacts Neurotoxins Pain Pattern Peptides Pharmaceutical Preparations Phylogenetic Analysis Physiological Positioning Attribute Process Protein Family Proteins Proteome Proteomics RNA Recruitment Activity Relative (related person)Relative Risks Research Set protein Specificity Spider Venoms Spiders Staging Taxon Tissues Toxin Transcript Trees United States National Institutes of Health Variant Venoms Work antimicrobial cDNA Library chemical function comparative drug discovery insight interest neurophysiology next generation sequencing novel response species difference sphingomyelin phosphodiesterase D therapy development

项目摘要

DESCRIPTION (provided by applicant): Animal venoms are complex mixtures of toxic components, including neurotoxins and cytotoxins with exquisite target specificity. Toxins with unique activities are of interest for drugs and neurophysiological research. Spiders, because of their venom complexity (200-1000 toxins in a single spider) and diversity (>44,500 described species!), likely contain the largest pool of toxin diversity of any venomous taxon, yet most major spider lineages are unexplored. Moreover, the field is in early stages of understanding evolutionary dynamics generating venom toxin diversity. With new availability of complete venom transcriptomes and the first spider genomes it is now possible to understand venom diversity and evolution at a new level of sophistication. Insight into evolutionary mechanisms influencing venoms can serve as a guide for toxin discovery and development of antivenom therapies. The goal of this project is to understand the phylogenetic scale at which there are significant differences in venom composition among spider taxa, and to infer mechanisms of evolution that contribute to these differences. This project continues comparative analyses of venomes in sicariid spiders and Haplogyne relatives. Sicariids include the notorious brown recluse whose bites cause dermonecrotic lesions and systemic effects in humans. There are ~130 described species and differences among them represent over ~120 million years of evolution since their most recent common ancestor. Evidence indicates the toxin responsible for dermonecrosis was recruited for venom function before the most recent common ancestor of the lineage. Sicariids are in the Haplogyne suborder within which, before this work, very little was known about venoms. Funded by NIH R15-GM-097696-01, cDNA transcriptomes for five sicariid taxa, three haplogynes and one outgroup were created and screened to produce nearly 5,000 venom gland transcripts. Combined with venom proteomics the minimal phylogenetic breadth of over 15 venom-expressed toxins was identified. However, these data are not comprehensive and missing toxins could reflect real absence, or artifacts of methods. There are three specific aims for this renewal: (1) to more thoroughly assess the phylogenetic distribution of venom toxin lineages across sicariids and haplogynes by adding next generation sequencing data; (2) to identify and analyze the gene families from which venom toxins were recruited by identifying non- venom expressed homologs in body transcriptomes and in the genome of Loxosceles reclusa; (3) to analyze evolutionary dynamics (duplications and selection) of individual lineages of toxins and compare them to identify parallel patterns in the evolution of toxins in the same venom functional complex. The toxins discovered in this work are a platform for discovery of natural components that contain unique neurophysiological activities. Preliminary data indicate discovery of divergent and unique toxins.

描述（由申请人提供）：动物毒液是有毒成分的复杂混合物，包括具有精确目标特异性的神经毒素和细胞毒素。具有独特活性的毒素引起药物和神经生理学研究的兴趣。由于蜘蛛毒液的复杂性（一只蜘蛛体内有 200-1000 种毒素）和多样性（超过 44,500 种描述的物种！），蜘蛛可能含有最大的毒素库任何有毒分类单元都具有多样性，但大多数主要蜘蛛谱系尚未被探索。此外，该领域正处于了解产生毒液毒素多样性的进化动力学的早期阶段。随着完整毒液转录组和第一个蜘蛛基因组的出现，现在可以在新的复杂水平上了解毒液的多样性和进化。深入了解影响毒液的进化机制可以作为毒素发现和抗蛇毒疗法开发的指南。该项目的目标是了解蜘蛛类群之间毒液成分存在显着差异的系统发育规模，并推断导致这些差异的进化机制。该项目继续对刀蛛和单腹蜘蛛的近缘蜘蛛的毒液进行比较分析。蜱虫包括臭名昭著的棕色隐士，其叮咬会导致人类皮肤坏死性病变和全身性影响。已描述的物种大约有 130 个，它们之间的差异代表了自它们最近的共同祖先以来大约 1.2 亿年的进化。有证据表明，导致皮肤坏死的毒素是在该谱系的最近共同祖先出现之前就被招募来发挥毒液功能的。锯齿虫属于单型亚目，在这项工作之前，人们对这个亚目的毒液知之甚少。在 NIH R15-GM-097696-01 的资助下，创建并筛选了 5 个 sicariid 类群、3 个单倍体和 1 个外类群的 cDNA 转录组，产生了近 5,000 个毒腺转录本。结合毒液蛋白质组学，确定了超过 15 种毒液表达毒素的最小系统发育宽度。然而，这些数据并不全面，缺失的毒素可能反映出真正的缺失或方法的伪影。此次更新有三个具体目标：（1）通过添加下一代测序数据，更彻底地评估跨蜘蛛纲和单倍体的毒液毒素谱系的系统发育分布； (2) 通过鉴定身体转录组和隐裂线鱼基因组中非毒液表达的同源物，鉴定和分析从中招募毒液毒素的基因家族； (3) 分析各个毒素谱系的进化动力学（重复和选择），并对它们进行比较，以确定同一毒液功能复合体中毒素进化的平行模式。这项工作中发现的毒素是发现含有独特神经生理活性的天然成分的平台。初步数据表明发现了不同且独特的毒素。