Abstract: The present disclosure generally relates to methods, systems and compositions for determining the presence or absence of prostate cancer in a subject, treating prostate cancer in a subject, and monitoring the efficacy of a treatment regimen for prostate cancer in a subject. The methods of the present disclosure comprise determining the expression levels and spatial locations of two or more analytes in a sample from a subject and correlating the expression levels and spatial locations to a prostate sample from a subject, thereby determining the presence or absence of prostate cancer, determining a treatment regimen for prostate cancer, or monitoring the efficacy of an existing treatment regimen in a subject undergoing treatment for prostate cancer.