Cahokia is the largest documented urban settlement in the pre-Columbian United States. Archaeological evidence suggests that the city, located near what is now East St. Louis, Illinois, began to rapidly expand starting around AD 1050. At its height, Cahokia extended across 1000 ha and included large plazas, timber palisade walls, and hundreds of monumental earthen mounds. Following several centuries of occupation, the city experienced a period of gradual abandonment from about AD 1200 to 1400. Here, we present geochemical data from a 1500-year-old sediment core from nearby Horseshoe Lake that records watershed impacts associated with the growth and decline of Cahokia. Sedimentary analysis shows a distinctive 24-cm-thick, gray, fine-grained layer formed between AD 1150 and 1220 and characterized by low carbonate δ13C, elevated sorbed metal concentrations, and higher organic matter δ15N. The deposition of this layer is contemporaneous with archaeological evidence of increased agricultural activity, earthen mound construction, and higher populations surrounding the lake. We hypothesize that these human impacts increased soil erosion, producing new sediment sources from deeper soil horizons, and shifted dissolved transport to the lake, producing lower carbonate δ13C values, higher concentrations of lead, copper, potassium, and aluminum, and increased δ15N, likely due to contributions of enriched nitrogen from sewage.