Project Summary/Abstract: African American (AA) individuals with prostate cancer (PCa) face significantly worse clinical outcome than their European American (EA) counterparts. There is a strong correlation between the stage of disease (localized versus metastatic) at diagnosis and long-term survival. While patients diagnosed with localized disease have a 99% 5-year survival, patients presenting with metastatic disease have significantly worse prospects, an about 32% 5-year survival. A minimally invasive, plasma based diagnostic method could significantly improve chances of detecting PCa at an early stage and thus reduce PCa related mortality. There are two, next generation sequencing (NGS)-based methods that likely have the required sensitivity and specificity of early detection of PCa from liquid, plasma biopsies. The first is based on the altered fragmentation profile of cancer derived cell free DNA (cfDNA). In this, whole genome sequencing (WGS) is applied to plasma derived cfDNA and the ratio of short to normal fragment size indicates the presence of cancer. The second method is based on a specific pattern of methylated DNA loci across the genome and combines cell-free methylated DNA immunoprecipitation with high throughput sequencing. We found that the altered cell free DNA fragmentation profile is a highly sensitive, robust indicator of the presence of metastatic prostate cancer. In this proposal we will investigate whether this method can identify cases when AA men present at diagnosis or at later time-point after post-primary treatment with metastatic PCa. We will also investigate whether a clinically useful sensitivity and specificity is retained as we analyze samples at earlier stages of disease. We will benchmark and optimize an experimental and computational protocol to detect prostate cancer at various stages based on the fragmentation profile of plasma derived cell free DNA. This will be applied to AA patients that presented with metastatic disease at diagnosis or later developed metastatic disease, to AA PCa patients with localized or locally advanced disease, and to AA PCa cases where sequential plasma was collected at various times, ranging from right before surgery to ten years before disease progression. We will determine the sensitivity and specificity of a fragmentomics profile-based method for early detection of PCa of various disease stages. Similarly, we will benchmark and optimize an experimental and computational protocol to detect prostate cancer at various stages based on cell free methylated DNA profiles. The next generation sequencing based methylation profiling of plasma derived cfDNA samples will be applied to AA PCa of various disease stages and we will determine the sensitivity and specificity of DNA methylation profile-based method for early detection of PCa of various disease stages. This will establish a non-invasive method for the early detection of PCa of AA men.