Project Summary/Abstract – Biomarker Reference Laboratory The proposed BCC will develop sensitive genomic and proteomic tests for the early detection of ovarian cancer in uterine lavage samples. The genomic component will assess genome-wide methylation, and the proteomic component will assay proteins using the proximity extension assays (PEAs) which provide relative quantitation between samples. The test will be optimized on samples from a training cohort. The methylation signal will be aggregated and optimized across the genome using machine learning approaches by the biomarker developmental lab (BDL). Optimization of the proteomic component will identify a custom panel of up to 21 proteins for absolute quantitation. The Biomarker Reference Laboratory (BRL) will work with Fulgent Genetics to ensure that optimized methylation signal and the custom panel of protein assays comply with CLIA/CAP standards and achieve analytical performance metrics specified by the BRL. The PI of BRL has extensive experience in ensuring assays in her clinical biochemistry lab serving the University Health Network, the largest in Canada, achieve clinical grade before analysis of patient samples are implemented in the lab’s workstream. The BRL will apply similar standards, agreed upon by all investigators in the BCC, to the methylation aggregated signal, the custom panel, and the classifier formed by combining the methylation signal with the protein abundances in the custom panel. This test using the classifier combining the genomic and proteomic signals, qualified with these standards, will give the highest probability of success for a test for the early detection of ovarian cancer when applied to uterine lavage samples from an independent validation cohort, and to cervical swabs as a readily accessible biospecimen with high patient acceptability.