Integrative Multidisciplinary Discovery Platform to Unlock Marine Natural Products Therapeutic Opportunities

项目总结 我们已经组建了一个完全以佛罗里达为中心的合作研究团队，他们拥有微生物方面的集体专业知识 天然产物化学和药理学，基因组学，细菌酶学，生物信息学，合成 生物学、化学合成、蓝藻和海绵化学生态学和系统发育。这个团队是 辅以一位外州的元基因组学和生物信息学整合专家。这个地理位置 位于美国大陆海洋生物多样性最大的州的专业知识集群提供了双重优势 受益和独特的机会来探索可能在以下方面拥有最大希望的系统 生物合成潜力：海洋蓝藻群落，由底栖丝状蓝藻组成， 与独特的微生物多样性有关，海绵及其相关的丰富和独特的微生物群在 当地的生物多样性热点。众所周知，由这些社区产生的化合物具有治疗作用 相关的化学空间，因此适合作为药物发现的起点。我们以有针对性的方式， 将从海绵相关基因获得高质量的(元)基因组和(元)转录组序列信息 使用最先进的测序技术的微生物群和蓝藻。我们将建设一个综合的、多方面的 组件平台，利用现有的生物信息学工具和新开发的基于人工智能的 工具为他们的基因组带来新的曙光，目标是识别新的生物合成基因簇，特别是 这些是目前的工具无法达到的，甚至是化学骨架。我们将对天然产品进行编码快递 通过使用我们战略上拥有的五种互补的合成生物学系统 在过去的几年里发展起来的。这些系统起源于五个共同关联的细菌门 海洋蓝藻和海绵样本涵盖了不同的遗传背景，预计 有效地将识别的各种生物的遗传信息转化为具有适当系统的化学物质 优化。我们将使用基于LC-MS的方法来评估和分析代谢物和表达谱 代谢组学和核磁共振，并对相关的新酶学进行表征。从化学物质中提取的天然产物 提取物和馏分文库以及通过我们的表达系统或化学合成生成的文库将是 在我们的多维筛选平台中进行测试，包括各种体外无偏见的表型分析 以及活体模型以及基于实验和计算靶点的功能分析。这种方法 将使我们能够从表达和未表达的基因中捕获一系列广泛的活动。精选生物活性 天然产物将通过化学合成进行规模化，并将制备生物探针和酶底物 分别用于深入的生物学研究和酶学研究。在以下情况下成功完成这些目标 我们现有的多学科研究团队应该提供有前景的治疗重要药物线索 并通过彻底探索海洋生物来工具化合物，同时解决当前主要的 未来十年天然产物药物发现的局限性。

PROJECT SUMMARY We have assembled an entirely Florida-centric collaborative research team with collective expertise in microbial natural products chemistry and pharmacology, genomics, bacterial enzymology, bioinformatics, synthetic biology, chemical synthesis, and cyanobacterial and sponge chemical ecology and phylogenetics. The team is complemented by an out-of-state expert in metagenomics and bioinformatics integration. This geographical cluster of expertise being in the state with the greatest marine biodiversity in the continental US provides a dual benefit and unique opportunity to explore systems that are likely to hold some of the most promise in terms of biosynthetic potential: marine cyanobacterial communities, consisting of benthic filamentous cyanobacteria that are associated with unique microbial diversity, and sponges and their associated rich and unique microbiome in a local hotspot of biodiversity. Compounds produced by these communities are known to cover therapeutically relevant chemical space and are therefore suited as starting points for drug discovery. In a targeted fashion, we will obtain high quality (meta)genome and (meta)transcriptome sequence information from sponge-associated microbiomes and cyanobacteria using state-of-the-art sequencing techniques. We will build an integrated, multi- component platform that leverages existing bioinformatics tools and newly developed artificial intelligence-based tools to shine new light at their genomes with the goal of identifying novel biosynthetic gene clusters, particularly those unattainable with current tools, and even chemical skeletons. We will express natural products encoded by the clusters by employing five types of complementary synthetic biology systems that we have strategically developed over the past several years. These systems originating from five bacterial phyla commonly associated with both marine cyanobacterial and sponge samples cover diverse genetic backgrounds and are expected to effectively translate the identified genetic information of a variety of organisms into chemicals with proper system optimization. We will evaluate and analyze metabolites and expression profiles using LC-MS-based metabolomics and NMR and characterize associated new enzymology. Natural products derived from chemical extract and fraction libraries and those generated through our expression systems or chemical synthesis will be tested in our multidimensional screening platform, consisting of unbiased phenotypic assays in various in vitro and in vivo models as well as experimental and computational target-based functional assays. This approach will enable us to capture a broad array of activities from expressed and unexpressed genes. Selected bioactive natural products will be scaled by chemical synthesis, and bioprobes and enzyme substrates will be prepared for in-depth biological studies and enzymology research, respectively. Successful completion of these aims by our established multidisciplinary investigator team should deliver promising therapeutically important drug leads and tool compounds through thoroughly exploring marine organisms while addressing the current major limitations of natural products drug discovery over the next ten years.