Integrative Venomics: Studies on venom composition, function and physiology of the venom gland system of non-front-fanged colubroids - using the western hognose snake (Heterodon nasicus) as an example

综合毒液组学：无前牙蛇毒腺系统的毒液成分、功能和生理学研究——以西部猪鼻蛇（Heterodon nasicus）为例

• 批准号: 540833593
• 负责人: Dr. Maik Damm
• 金额: --
• 依托单位: Institut für Insektenbiotechnologie
• 依托单位国家: 德国
• 项目类别: WBP Position
• 资助国家: 德国
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/540833593?language=en
• 关键词: Integrative; Venomics; Studies; venom; composition

Snakebites are of worldwide medical significance, with over 5 million cases per year. The most clinically relevant snakebite envenomation originate from vipers (Viperidae) and elapids (Elapidae), which are grouped together as front-fanged snakes (FFS) due to their dentition. The deciphering of snake venoms, which consist of proteins and peptides, plays a major role in the development of antidotes, drugs, and the understanding of venoms in general. However, current knowledge largely originates only on those species that are medically relevant. This anthropocentric approach is based on a small taxonomic intersection, as recent work has shown that many more snakes are venomous than previously thought. These non-front-fanged colubroids (NFFC) far exceed the 750 FFS species, with >2500 species. Few NFFC venoms have ever been studied and have shown significant differences from the well-studied FFS venoms. And yet their potential has hardly been explored or exploited. The aim of this project is to gain understanding of the venom composition, function, and physiology of the venom system of NFFC. I will examine these points using the western hognose snake (Heterodon nasicus) as an example, with its posterior fangs. Although they are kept as pets, which offers advantages for venom milking as well as conservation and the Nagoya protocols, case reports have shown striking signs of envenomation for decades. Symptoms range from localized pain and massive blistering to thrombocytopenia. However, the venom has never been studied in detail or with modern analytical methods. The workflow is divided into three parts and considers the main aspects of proteomic composition, toxin production in tissues and physiological effects of the venom. In the first part, the individual toxins and components are identified using state-of-the-art Snake Venomics protocols. Using high-resolution mass spectrometry, the toxin profile is determined holistically (bottom-up, top-down, peptidomics). In the second part, the distribution of toxins within a western hognose snake venom gland is determined for the first time of a NFFC by mass spectrometry imaging (MALDI Imaging, Spatial Venomics) and put into a direct histological context. Final will examine venom function, using bioassays to characterize the exochemistry of the NFFC snake venom. They include assays for blood coagulation, cytotoxicity, enzyme activity, and potential antimicrobial activities. This will contribute to the basic understanding of western hognose snake biology and serve as a model to better understand venom plasticity in NFFC snakes worldwide.

蛇咬伤在世界范围内具有重要的医学意义，每年发生超过 500 万例。临床上最相关的蛇咬伤源自毒蛇（Viperidae）和蛇蛇（Elapidae），由于它们的齿列，它们被归为前牙蛇（FFS）。蛇毒由蛋白质和肽组成，其破译在解毒剂、药物的开发以及对毒液的总体理解中发挥着重要作用。然而，当前的知识很大程度上仅源于那些与医学相关的物种。这种以人类为中心的方法是基于一个小的分类交叉点，因为最近的研究表明，有毒的蛇比之前想象的要多得多。这些无前牙疣类动物 (NFFC) 远远超过 FFS 的 750 种，超过 2500 种。很少有 NFFC 毒液被研究过，并且与经过充分研究的 FFS 毒液显示出显着差异。然而，它们的潜力却几乎没有被探索或利用。该项目的目的是了解 NFFC 毒液系统的毒液成分、功能和生理学。我将以西部猪鼻蛇（Heterodon nasicus）及其后尖牙为例来研究这些要点。尽管它们被作为宠物饲养，这为毒液挤奶以及保护和名古屋协议提供了优势，但数十年来病例报告显示出明显的中毒迹象。症状范围从局部疼痛、大面积水疱到血小板减少。然而，从未对毒液进行过详细研究或使用现代分析方法。工作流程分为三个部分，考虑了蛋白质组组成、组织中毒素产生和毒液生理效应的主要方面。在第一部分中，使用最先进的蛇毒组学协议来识别单个毒素和成分。使用高分辨率质谱法，全面确定毒素谱（自下而上、自上而下、肽组学）。在第二部分中，首次通过质谱成像（MALDI 成像，空间毒液学）确定了 NFFC 中西部猪鼻蛇毒腺内毒素的分布，并将其置于直接组织学背景中。最后将检查毒液功能，使用生物测定来表征 NFFC 蛇毒的放出化学。它们包括凝血、细胞毒性、酶活性和潜在抗菌活性的测定。这将有助于对西方猪鼻蛇生物学的基本了解，并作为更好地了解全球 NFFC 蛇毒液可塑性的模型。