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<div>AbstractPurpose:<p>ctDNA is a novel technique extensively studied in solid tumors, although not currently well defined in endometrial cancer.</p>Experimental Design:<p>A de-identified retrospective analysis of 1,988 patients with advanced/recurrent endometrial cancer was performed. In addition, an analysis of a real-world evidence cohort was completed (<i>n</i> = 1,266). Patients underwent ctDNA testing using Guardant360 during routine clinical care. The objective was to describe and assess molecular landscape using ctDNA.</p>Results:<p>Among 1,988 ctDNA samples, at least one somatic alteration was detected in 91.6% (<i>n</i> = 1,821). Most frequently altered genes were <i>TP53</i> (64%), <i>PIK3CA</i> (29%), <i>PTEN</i> (25%), <i>ARID1A</i> (20%), and <i>KRAS</i> (14%). Overall, 18.5% had amplifications, with the majority identified in <i>CCNE1</i> (40.9%), <i>PIK3CA</i> (22%), and <i>EGFR</i> (19.3%). From the real-world evidence cohort, those with <i>TP53</i> mutations had a worse overall survival (OS) versus those without <i>TP53</i> mutations (<i>P</i> = 0.02) and those with <i>TP53</i> comutations had an inferior OS in comparison with <i>TP53</i>-mutated only (<i>P</i> = 0.016). Amongst these, patients with a <i>PIK3CA</i> comutation (<i>P</i> = 0.012) and <i>CCNE1</i> amplification (<i>P</i> = 0.01) had an inferior OS compared with those with only <i>TP53</i> mutations. Fifty-seven patients with newly diagnosed endometrial cancer had at least two serial ctDNA samples showing evolution in detected variants compared with baseline samples, with <i>TP53</i> being the most frequent change.</p>Conclusions:<p>This study is one of the largest cohorts of ctDNA currently reported in endometrial cancer. The presence of <i>TP53</i> mutation and other comutations detected by ctDNA have a negative effect on outcomes. This report suggests that ctDNA analysis is feasible and could become a useful biomarker for endometrial cancer.</p></div>