The overarching goal of this Program Project proposal is to discover natural products that can serve as anticancer drug leads from diverse natural sources. Project 2, located at the College of Pharmacy, University of Illinois at Chicago (UIC), is comprised of botanical, chemical and biological elements. The primary goals for Project 2 are the discovery of new lead anticancer compounds from cyanobacteria and providing tropical plant material for Project 1, housed at The Ohio State University (OSU). The underlying hypothesis for Project 2 is that the diverse natural product structures from cultured non-marine cyanobacteria and tropical plants will be a rich source for anticancer leads. To achieve the stated goals the specific aims for Project 2 are as follows: 1) to obtain, culture, extract, and prepare fractions of cyanobacterial strains, and to sequence the genome of selected strains. We will obtain 100 cyanobacterial strains per year. The strain collection will focus on samples collected in the Upper Midwest and Great Lakes regions of the U.S. Each strain will be cultured, extracted, fractionated, and provided for biological evaluation in the biological test systems available at Core 1 and Project 1 as well as our external providers of bioassays. As a complementary and integrated approach, we will sequence and genome mine the genomes of 20 strains per year and strains will be selected for sequencing based on a) promising biological activity; and b) prioritization based on chemotaxonomy or PCR screen. 2) To isolate and determine the structures of active cyanobacterial metabolites, and to improve yields of lead compounds. Extracts showing promising activity will be fractionated using activity-monitored fractionation to obtain pure natural product(s). The structurally characterized, active isolates will be extensively evaluated in the biological test systems available to the program. We will re-isolate larger amounts of any candidate compound. In order to improve yields, when necessary, we will evaluate different cultivation conditions, and explore heterologous expression of the biosynthetic gene cluster of lead compounds, which will serve to provide an alternative source for production. 3) To acquire all plant materials that will be required by the proposed Program Project. We will obtain 150 fully documented and identified plant samples per year from selected countries of the world. The dried and milled plant material will be provided to Project 1 for further processing. We will also re-collect plant material for larger-scale isolation of active compounds requiring more detailed biological evaluation as needed.