title: Integration of high-content screening and untargeted metabolomics for comprehensive functional annotation of natural product libraries.
abstract: Traditional natural products discovery using a combination of live/dead screening followed by iterative bioassay-guided fractionation affords no information about compound structure or mode of action until late in the discovery process. This leads to high rates of rediscovery and low probabilities of finding compounds with unique biological and/or chemical properties. By integrating image-based phenotypic screening in HeLa cells with high-resolution untargeted metabolomics analysis, we have developed a new platform, termed Compound Activity Mapping, that is capable of directly predicting the identities and modes of action of bioactive constituents for any complex natural product extract library. This new tool can be used to rapidly identify novel bioactive constituents and provide predictions of compound modes of action directly from primary screening data. This approach inverts the natural products discovery process from the existing "grind and find" model to a targeted, hypothesis-driven discovery model where the chemical features and biological function of bioactive metabolites are known early in the screening workflow, and lead compounds can be rationally selected based on biological and/or chemical novelty. We demonstrate the utility of the Compound Activity Mapping platform by combining 10,977 mass spectral features and 58,032 biological measurements from a library of 234 natural products extracts and integrating these two datasets to identify 13 clusters of fractions containing 11 known compound families and four new compounds. Using Compound Activity Mapping we discovered the quinocinnolinomycins, a new family of natural products with a unique carbon skeleton that cause endoplasmic reticulum stress.
authors.affiliation: Department of Chemistry and Biochemistry, University of California, Santa Cruz, CA 95064.
authors.forename: Kenji L
authors.initials: KL
authors.lastname: Kurita
authors.affiliation: Department of Chemistry and Biochemistry, University of California, Santa Cruz, CA 95064.
authors.forename: Emerson
authors.initials: E
authors.lastname: Glassey
authors.affiliation: Department of Chemistry and Biochemistry, University of California, Santa Cruz, CA 95064 rliningt@sfu.ca.
authors.forename: Roger G
authors.initials: RG
authors.lastname: Linington
mesh_terms: D001688:Biological Products / Q000032:analysis*; D066289:Data Curation / Q000379:methods*; D019992:Databases as Topic; D055808:Drug Discovery / Q000379:methods*; D006367:HeLa Cells; D006801:Humans; D007091:Image Processing, Computer-Assisted / Q000379:methods; D055432:Metabolomics / Q000379:methods* / Q000592:standards
chemical_list: D001688:Biological Products
keywords: bioactive small molecules; image-based screening; informatics; metabolomics; natural products