Abstract 5163: A high sensitivity, tumor-informed liquid biopsy platform, designed to detect minimal residual disease at part per million resolution
Authors: Boyle Sean Michael , Bartha Gabor , Lyle John , Harris Jason , Northcott Josette , Norton Dan , Pyke Rachel Marty , Navarro Fabio C. P. , Stram Alexander , Haudenschild Christian , Santiago Rose , Li Robin , Nelson Chris , Huo Yelia , Chinnappa Manju , Zhang Qi , Hsu Lloyd , West John , Chen Richard O. 
Published: June 2022

Tumor-informed liquid biopsy approaches have proven promising for detecting minimal residual disease (MRD) and recurrence of cancer following surgical resection or other therapy. However, current liquid biopsy MRD assays typically detect ctDNA in a range above 30 to 300 parts per million (PPM), leaving a significant fraction of MRD cases undetected, particularly soon after surgery and in early stage cancers where ctDNA can be at very low levels. To address this, we have developed NeXT Personal™, a tumor-informed liquid biopsy assay that achieves sensitivity down to 1 PPM, therefore enabling earlier detection of MRD and recurrence.

NeXT Personal leverages tumor/normal whole genome sequencing to design personalized MRD liquid biopsy panels for each patient. The panel is composed of &gt;1,200 somatic tumor variants enabling higher sensitivity MRD detection in plasma through tracking of larger numbers of high quality and lower noise variants. This allows the platform to achieve high sensitivity across cancer types and stages, including early stage cancers and low mutational burden tumors, utilizing ~4 mL of plasma. Two independent methods were used to establish utility and performance: a proprietary cell-line media system, and well-characterized matched tumor-normal-plasma patient samples. Samples were serially diluted to &lt;1 PPM, with replicates used to confirm performance. Digital droplet polymerase chain reaction (ddPCR) was used to orthogonally validate platform performance to the limit of detection (LOD) of ddPCR.

Characterization of MRD LOD in three cell-line media systems, HCC1143, HCC38, and HCC1937, yielded accurate and reproducible detection of signal across a broad range of concentrations, to a lower limit of 1-2 PPM. We then used our platform to characterize MRD LOD in a set of serially diluted patient samples, demonstrating sensitivity down to as low as 1 PPM, with high specificity in normal control samples. Finally, we demonstrated the performance of NeXT Personal with matched tumor-normal-plasma patient samples (8 different cancer types, stages II-IV). In this series, NeXT Personal detected cancers down to 0.8 PPM with high specificity demonstrated across a set of healthy normal donor samples. We estimate that ~50% of the cases in this set of patients would not have been detected by other commercially available liquid biopsy MRD platforms.

NeXT Personal achieved highly sensitive and specific MRD detection, reproducibly demonstrating a LOD down to 1 PPM in different cancer types and cell line dilutions, representing approximately 10 to 100 times higher sensitivity than other liquid biopsy MRD approaches. The high sensitivity of NeXT Personal potentially enables MRD detection across a broad variety of cancers and stages, including typically challenging early stage, low mutational burden, and low-shedding cancers.

Citation Format: Sean Michael Boyle, Gabor Bartha, John Lyle, Jason Harris, Josette Northcott, Dan Norton, Rachel Marty Pyke, Fabio C. P. Navarro, Alexander Stram, Christian Haudenschild, Rose Santiago, Robin Li, Chris Nelson, Yelia Huo, Manju Chinnappa, Qi Zhang, Lloyd Hsu, John West, Richard O. Chen. A high sensitivity, tumor-informed liquid biopsy platform, designed to detect minimal residual disease at part per million resolution [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5163.