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PURPOSEDetection of persistent circulating tumor DNA (ctDNA) after curative-intent surgery can identify patients with minimal residual disease (MRD) who will ultimately recur. Most ctDNA MRD assays require tumor sequencing to identify tumor-derived mutations to facilitate ctDNA detection, requiring tumor and blood. We evaluated a plasma-only ctDNA assay integrating genomic and epigenomic cancer signatures to enable tumor-uninformed MRD detection.EXPERIMENTAL DESIGN252 prospective serial plasma specimens from 103 colorectal cancer (CRC) patients undergoing curative intent surgery were analyzed and correlated with recurrence.RESULTSOf 103 patients, 84 (stage I (9.5%), II (23.8%), III (47.6%), IV (19%)) had evaluable plasma drawn post-completion of definitive therapy, defined as surgery only (n=39) or completion of adjuvant therapy (n=45). In "landmark" plasma drawn one-month (median 31.5 days) post-definitive therapy and >1 year follow up, 15 patients had detectable ctDNA, and all 15 recurred (positive predictive value (PPV) 100%, hazard ratio 11.28 (p<0.0001)). Of 49 patients without detectable ctDNA at the landmark timepoint, 12 (24.5%) recurred. Landmark recurrence sensitivity and specificity were 55.6% and 100%. Incorporating serial longitudinal and surveillance (drawn within 4-months of recurrence) samples, sensitivity improved to 69% and 91%. Integrating epigenomic signatures increased sensitivity by 25-36% versus genomic alterations alone. Notably, standard serum carcinoembryonic antigen (CEA) levels did not predict recurrence (hazard ratio 1.84 (p=0.18); PPV=53.9%).CONCLUSIONSPlasma-only MRD detection demonstrated favorable sensitivity and specificity for recurrence, comparable to tumor-informed approaches. Integrating analysis of epigenomic and genomic alterations enhanced sensitivity. These findings support the potential clinical utility of plasma-only ctDNA MRD detection.

Minimal Residual Disease Detection using a Plasma-only Circulating Tumor DNA Assay in Patients with Colorectal Cancer