Automation of a Blood Test for Methylation Markers to Reliably Predict Response to Therapy and Prognosis of Outcome in Patients with Metastatic Breast Cancer
Abstract
Reliable and rapid tests for the prediction of response to an initiated therapy, and prognostication of the outcome of metastatic breast cancer are sorely needed. Our goal is to develop a 3-4-hour, automated blood test based on quantitative, multiplexed assessment of methylated, tumor specific, cell-free DNA in the blood. We have completed the goals set out in the SOW: that is, to devise the PCR conditions in which 6 genes (5 methylated genes plus a control ACTB) could be amplified using 6 different fluorophores in the same reaction, using templates of 200-600 copies of methylated DNA in 1 ml of plasma, while producing no false signals in the absence of methylated template DNA. More improvements to the assay and completion of training, and test sets showed that the test is performing at 95 percent sensitivity and 92 percent specificity with a ROC/AUC of 0.909. Validation analysis was accomplished in samples from a prospective TBCRC005 trial. At week 4 MBC patients with high cumulative methylation (CM) had a significantly shorter median PFS (2.88 months v 6.60 months, p = 0.001) and OS (14.52 months v 22.44 months, p=0.005) compared to those with low CM (accepted for publication). Blood tests that function with accuracy will minimize morbidity from ineffective therapy, reduce costs from additional imaging studies, and improve clinical outcomes.
Document Details
- Document Type
- Technical Report
- Publication Date
- Apr 01, 2023
- Accession Number
- AD1212992
Entities
People
- Saraswati Sukumar
Organizations
- Johns Hopkins University