Introduction: The PREDICT DNA trial is the first prospective, multi-center study aimed at validating cell-free plasma derived circulating tumor DNA (ctDNA) as a biomarker for treatment response and recurrence in early stage, triple-negative or HER2-positive (any hormone receptor status) breast cancer. Its primary aim is to determine the negative predictive value (NPV) of the absence of ctDNA after neoadjuvant therapy (NAT) for the achievement of pathologic complete response (pCR). This study has met its accrual goals and results of the overall trial are anticipated within the next year. The initial PREDICT DNA study design stipulated that tumor specific mutations (TSMs) to be tracked in blood would be identified by next gen sequencing (NGS) of tumor biopsy tissue. A disadvantage of this design is the dependence on adequate biopsy tissue. Recently, the advent of Safe-SeqS technology has enabled robust detection of rare variants using NGS with a sensitivity of approximately 0.05% mutant allele fraction. We employed these new NGS methods to pilot a novel tissue-independent approach to ctDNA detection and monitoring. Objective: The primary objective of this pilot study was to determine whether ultrasensitive NGS using a targeted cancer mutation panel can identify TSMs in ctDNA of early-stage breast cancer patients without the use of biopsy tissue. Methods: The PREDICT DNA trial enrolled 228 women from 22 sites with stage II/III breast cancer for whom standard neoadjuvant therapy was planned. Of these, 58 patients had matched pre-and post-NAT samples available for analysis at the time of this pilot. All pre-NAT samples were analyzed for the presence of TSMs using Sysmex Inostics’ SafeSEQ. Patients with detectable ctDNA before NAT were also evaluated for residual ctDNA after completion of NAT but prior to surgery. Five samples were also tested by digital PCR (BEAMing) for cross-platform comparison. Results: TSMs in ctDNA were identified in 29 of 58 patients (50%) prior to NAT. Of pre-NAT ctDNA(+) patients, TSMs were detected in TP53 (90%) and PIK3CA (10%); three patients (10%) were found to have 2 TSMs. Concordance between SafeSEQ and BEAMing was 100% in five samples tested [3 ctDNA(+), 2 ctDNA(-)]. Of 29 ctDNA(+) patients, 24 (83%) demonstrated reduction or elimination of detectable ctDNA following neoadjuvant therapy, with 16 (55%) converting to ctDNA(-). Conclusion: Identification of TSMs in the plasma of early-stage breast cancer patients without the need for biopsy tissue is feasible using a SafeSEQ cancer mutation panel. Further measures to improve the sensitivity of pre-treatment TSM analysis, such as increased plasma volume input and comprehensive TP53 mutational analysis are currently under investigation. Correlations between clinicopathologic factors with ctDNA detection and burden, as well as the NPV of post-NAT ctDNA for pCR and residual cancer burden, will be reported at the time of abstract presentation.

NH and HP contributed equally to this work.

Citation Format: Natasha Hunter, Heather Parsons, Alexander Sherry, Daniel Shinn, Dong Ho Shin, Alex Cole, Giovanni Cragnotti, Taylor Groginski, Margaret Leathers, Andrea L Richardson, Pedram Argani, Antonio Wolff, Leslie Cope, Dan Edelstein, Frank Holtrup, Hilary Sloane, Bapsi Chakravarthy, Vered Stearns, Ben H Park. TBCRC 040: Pathologic response evaluation and detection in circulating tumor DNA (PREDICT DNA): Initial results piloting a tissue-biopsy independent method of identifying and monitoring tumor-specific mutations in early stage breast cancer [abstract]. In: Proceedings of the 2019 San Antonio Breast Cancer Symposium; 2019 …