Discovery of natural products that modulate signaling in patient-derived cells

Multiplexed Activity Metabolomics, which correlates single cell assays with spectrally defined chromatographic arrays to identify bioactive metabolomic features prior to isolation. Two experimental case studies using this workflow with cave-microbe extracts identified new molecular phenotypes of the pyridine-pyrrolidine alkaloid siderochelin and the pyrrolopyrrole-functionalized anthracycline isoquinocycline B. Finally, we investigated the effects of anthracycline functionalization on human primary cells using single cell mass cytometry to understand if changes in anthracycline aglycon structure and glycosylation impacted cell selectivity. Despite sharing an anthracyclinone pharmacophore core, variants differentially impacted responses of leukemia cell populations within and among acute myeloid leukemia (AML) patient samples. This suggests pharmacophore assumptions may not guide assessment of therapeutic potential for anthracyclines and that modest structural differences can elicit marked changes in cellular function in different patients. Taken together, the depth of information afforded by single cell molecular phenotype-based discovery and mass cytometry deep cell profiling provides new patient-level insight into biological mechanisms of new and previously discovered molecules that may find expanded use in the clinic.