Project 1 at The Ohio State University (OSU) is comprised of taxonomic authentication, isolation chemistry, and biological testing components. Specific Aim 1 will involve receiving extracts of bioactive authenticated tropical plant materials from Project 2 (University of Illinois at Chicago, UIC) for phytochemical work-up in Aim 2, and extracting with solvents taxonomically authenticated U.S. coastal lichens and their cultured fungal associates (mycobionts), with these fungi identified from their 18S rDNA sequence. Specific Aim 2 will deal with (a) LC-MS dereplication studies on active leads following preliminary screening (in order to prioritize samples for activity-guided fractionation); (b) purification procedures using chromatographic methods (guided by both in-house bioassays and those elsewhere in the program project); (c) compound structure elucidation, and (d) scale-up isolation, respectively. The structures of bioactive compounds will be determined using modern spectroscopic methods, backed up by chemical transformations and X-ray crystallography [work to be performed Projects 1, 2, and the medicinal chemistry portion of Core 2 (OSU)], where necessary. In Specific Aim 3, plant, lichen, and fungal extracts will be subjected to primary screening against a small panel of cancer cell lines at Core 1 (UIC). The Project 1 bioassays will involve three target-based assays [viz., nuclear factor-κB (NF-κB), and nuclear receptor (Rev-Erb)], and mitochondrial transmembrane potential (MTP) testing, followed by preliminary mechanistic evaluations. Moreover, selected pure compounds will be evaluated for potential targeting to DNA topoisomerase IIα (TOP2α), as well as testing for DNA damage, cytotoxicity, pro-oxidant activity and glutathione-level perturbations. These assays will be available also for the testing of compounds isolated in Projects 2 and 3 and Core 2. A strong collaborations has been established to test promising compounds from Projects 1-3 at the Nationwide Children’s Hospital, Columbus, OH [NF2 (neurofibromatosis type 2)-associated vestibular schwannomas and meningiomas]. In addition, selected compounds will be submitted for formulation, stability, solubility, metabolism and pilot pharmacokinetic studies in Core 2 (OSU), and also tested in hollow fiber and xenograft in vivo assays in Core 1 (UIC), and in cancer-related in vitro bioassays at NCATS, NIH. Assay refinement in Project 1 will be conducted in collaboration with the biostatistics component of Core A (OSU).