Life history-guided drug discovery from venomous marine snails

摘要 柯诺总科的有毒海洋蜗牛通过注射一种复合体来捕获猎物 核糖体合成肽的混合物，经过广泛的翻译后修饰。 这些柯多肽针对猎物神经、内分泌和神经中的受体和离子通道。 感官系统具有显著的效力和特异性。由于他们的多样性和目标 选择性，已经成为离子通道研究和AS的宝贵工具 治疗学。使用锥形蜗牛毒液作为药物发现来源的理由是 在锥形蜗牛的猎物中表达的许多分子靶标的同源基因也在人类中发现 它们与多种生理紊乱有关，包括炎症、癫痫、 神经性疼痛和糖尿病。我的团队最近的几项发现现在证明了 大约700种锥形蜗牛中的每一种都会产生一套独特的螺杆肽，这些蛋白经过微调 寻找猎物体内的一组特定受体。因此，这笔赠款的中心假设是这种药物 通过对许多毒液成分进行测序和表征，可以最大限度地扩大发现 来自不同血统的锥形蜗牛的物种，包括那些引起不同生理反应的物种 它们猎物的终结点。这是一种高度创新的方法，因为它充分利用了 在这些动物中进化出的独特捕食策略：导致快速瘫痪的物种 在猎物中可能表达针对神经肌肉连接和痛觉回路的毒素 而那些导致活动不足和镇静的人更有可能进化出 瞄准感官和内分泌系统。我们的初步研究已经确定了几个 独一无二的药物用于治疗糖尿病，这种疾病已被公认为全球 流行和疼痛是目前阿片类药物流行的主要原因。这项提议将使我们能够 有效地扩展这些有希望的初始工作。本项目的具体目标是(目标1) 进行大规模的、以进化为导向的收集和下一代测序工作 来自所有~50个主要血统的锥形蜗牛的毒液，(目标2)开发一种创新的计算 管道，分类毒液模块，以分析这些大型测序数据集，和(目标3) 使用分级的、数据驱动的选择过程来确定最有希望的药物的药理学特征 来自这些大型数据集的新型毒素。我们还将寻求鉴定和鉴定刀鱼多肽 生物合成途径。这样做将改进生产的合成和重组手段 用于功能研究的蜈蚣多肽。预期结果是显著的。我们将提供一个 用于药物发现的计算流水线，将导致许多新的类别的识别 将为科学发现和药物提供动力的椰子肽及其生物合成酶 未来几十年的发展活动。

SUMMARY Venomous marine snails in the superfamily Conoidea capture their prey by injecting a complex mixture of ribosomally-synthesized peptides that undergo extensive post-translational modification. These conopeptides target receptors and ion channels in the prey's nervous, endocrine and sensory system with remarkable potency and specificity. Owing to their diversity and target selectivity, conopeptides have become invaluable tools for ion channel research and as therapeutics. The rationale of using cone snail venoms as a source for drug discovery is that homologs of many molecular targets expressed in the prey of cone snails are also found in humans where they are implicated in diverse physiological disorders, including inflammation, epilepsy, neuropathic pain and diabetes. Several recent discoveries made in my group now demonstrate that each of the ~700 cone snail species produces a distinct set of conopeptides that are finely tuned for a specific set of receptors in its prey. Thus, the central hypothesis of this grant is that drug discovery can be maximized by sequencing and characterizing the venom composition of many species from diverse lineages of cone snails, including those that induce diverse physiological endpoints in their prey. This is a highly innovative approach because it takes full advantage of the unique strategies that evolved in these animals for prey capture: species that induce rapid paralysis in their prey are likely to express toxins that target the neuromuscular junction and pain circuits whereas those that induce hypoactivity and sedation are more likely to have evolved toxins that target the sensory and endocrine system. Our preliminary research has already identified several unique drug leads for the treatment of diabetes, a disease that has been recognized as a global epidemic, and pain, a leading cause for the current opioid epidemic. This proposal will enable us to efficiently scale these promising initial efforts. The specific aims of this project are (Aim 1) to undertake a large-scale, evolution-guided collection and next-generation sequencing effort of venoms from all ~50 major lineages of cone snails, (Aim 2) to develop an innovative computational pipeline, the Taxonomer Venoms Module, to analyze these large sequencing datasets, and (Aim 3) to use a tiered, data-driven selection process to pharmacologically characterize the most promising novel toxins from these large datasets. We will also seek to identify and characterize conopeptide biosynthetic pathways. Doing so will improve synthetic and recombinant means for production of conopeptides for functional studies. The expected outcomes are significant. We will provide a computational pipeline for drug discovery that will lead to the identification of many novel classes of conopeptides and their biosynthetic enzymes that will fuel scientific discovery and drug development activities for decades to come.