Rapid discovery of thousands of intact biosynthetic gene pathways for bioactive natural product compounds from un-sequenced filamentous fungi using a novel FAC-NGS tool

Project Summary Abstract The goal of this project is to develop a highly rigorous technology platform utilizing lichen and unsequenced fungi to revive the discovery pipeline for fungal derived therapeutics capable of treating chronic infections and health conditionsThe emergence of drug resistant microbesthe diminishing supply of novel classes of antibioticsand the dramatic reduction in discovery and development of natural productsNPsalso termed secondary metabolitesSMsand other small molecule compounds from bacteria and fungi for anti infectiveanti proliferation and antiinflammation agents sinces have amplified a serious public health concern championed by the CDC and WHOWe posit that the revival of large scale drug discovery pipelines using underexploited microbes including lichen fungal symbionts and un sequenced fungi will provide a new cadre of novel drug leads and solutions towards the antibiotic resistance crisisFungal SMs are also important sources of anticancer compounds among other widespread clinical usesHoweveronlyor less of filamentous fungi have been sequenced and high throughput sequencing has shown that only aboutof fungal SM biosynthetic gene clustersBGCsare expressed under laboratory conditionsThereforerevolutionary technologies and tools are urgently needed to discover and more effectively dissect the biosynthesis of fungal SMs in order to more efficiently access novel fungal metabolites as potential pharmaceutical agentsRecentlywe developed a novel fungal artificial chromosome mass spectrometryFAC MSmethod that allows the direct captureheterologous expression and chemical analysis of an entire set of large intact SM BGCs from sequenced fungi as shown in Clevenger et alNatChemBiolWe have also shown that shuttle bacterial artificial chromosomeBACtechnology combined with BAC poolingindexingand next gen sequencingNGScan achievekb linked sequencing and assemblyThereforein this projectwe will develop an innovative fungal technology platform by integrating FAC MS with BAC NGS sequencing and bioactivity profiling to prove the concept that novel bioactive SMs can be captured from hard to growe glichen fungal symbiontsPhase Iand un sequenced fungiPhase IIThis technology should improve fungal SM discoveryfold and result in the discovery of at leastnovel antimicrobial drug leads from lichen and un sequenced fungi Project Narrative The goal of this project is to develop a rigorous technology platform utilizing lichen and unsequenced fungi to revive the discovery pipeline for fungal derived therapeutics capable of treating chronic infections and other health conditionsFungal natural productssome naturally possessing antimicrobial activityare a critical part of the solution to the current antibioticresistance crisisWe have developed a comprehensive technology platform capable of scalable drug discovery from filamentous fungi allowing us to tap into this vast undiscovered natural product pool