Abstract 913: Early detection of metastatic relapse and monitoring of therapeutic efficacy by ultra-deep sequencing of plasma cell-free DNA in patients with urothelial bladder carcinoma

Abstract

Background Circulating tumor DNA (ctDNA) analysis is an emerging field for diagnosis, monitoring of treatment efficacy and detection of disease recurrence in various cancers. In bladder cancer, previous work has demonstrated prognostic value of ctDNA for early stage cancers and early detection of recurrence following radical cystectomy (CX), however more extensive investigation of the potential impact of ctDNA analysis on clinical decision making remains to be carried out. Methods A total of 68 patients with localized advanced bladder cancer to be treated with neoadjuvant chemotherapy (NAC) and CX were prospectively included. We performed RNA-Seq to identify RNA subtypes and whole exome sequencing to identify somatic mutations for ctDNA analysis. Based on allele frequency and sequence context, 16 somatic mutations were selected for each patient for multiplex PCR and subsequent ultra-deep sequencing (median coverage of 114,000x). The sample-level analytical sensitivity was determined to be >95% at 0.01% ctDNA concentration level when at least 2/16 mutations were detected. The protocol was applied to detect ctDNA in 618 blood samples procured at diagnosis, during NAC, before CX and during surveillance following CX. Results The presence of ctDNA was highly prognostic already at diagnosis before initiation of NAC (HR=31.7, p=0.001). Analysis of ctDNA dynamics during NAC revealed that clearance of ctDNA was associated with pathological downstaging (≤TaN0 at CX) and reduced disease recurrence. Conversely, continually detectable ctDNA during NAC was associated with lack of pathological downstaging and later disease recurrence (p=0.023). Our results suggest that ctDNA dynamics during NAC may offer superior predictive value compared to clinical parameters and tumor centric molecular markers such as DNA damage response mutations and RNA subtypes. Following CX, ctDNA analysis identified 92% of metastatic relapse (100% specificity) during disease monitoring (100% positive- and 98% negative predictive value). The positive lead-time over radiographic imaging was 100 days on average (range: -17-245 days). Conclusions ctDNA analysis of blood samples procured throughout the disease course of patients with bladder cancer demonstrates novel evidence of early risk stratification and monitoring of treatment efficacy, which adds to the already strong evidence of ctDNA-based recurrence detection. Collectively, this paves the way for novel ctDNA-based clinical trials. Citation Format: Emil Christensen, Karin Birkenkamp-Demtröder, Himanshu Sethi, Svetlana Shchegrova, Raheleh Salari, Iver Nordentoft, Hsin-Ta Wu, Michael Knudsen, Philippe Lamy, Sia V. Lindskrog, Ann Taber, Mustafa Balcioglu, Søren Vang, Zoe Assaf, Shruti Sharma, Antony S. Tin, Ramya Srinivasan, Dina Hafez, Thomas Reinert, Samantha Navarro, Alexander Olson, Rosie Ram, Scott Dashner, Matthew Rabinowitz, Paul Billings, Styrmir Sigurjonsson, Claus L. Andersen, Ryan Swenerton, Alexey Aleshin, Bernhard G. Zimmermann, Mads Agerbæk, Cheng-Ho J. Lin, Jørgen B. Jensen, Lars Dyrskjøt. Early detection of metastatic relapse and monitoring of therapeutic efficacy by ultra-deep sequencing of plasma cell-free DNA in patients with urothelial bladder carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 913.

Plasma Circulating Tumor DNA Sequencing Predicts Minimal Residual Disease in Resectable Esophageal Squamous Cell Carcinoma

“…Pretreatment ctDNA levels also appear to be prognostic and help predict the dynamics of durvalumab treatment in 16 advanced tumor types as reported in a previous study (45). In patients with urothelial bladder carcinoma, the ctDNA-NGS approach could predict metastatic relapse and monitor therapeutic efficacy (46). Furthermore, plasma ctDNA is a potential biomarker for multiple cancers with high sensitivity and specificity (47)(48)(49)(50).…”

Biomarker Development Using Liquid Biopsy in Hepatocellular Carcinoma

“…There is evidence of its utility for predicting recurrence in both lung 72 and colorectal cancer 42 and is commercially available for colon, breast, lung, and bladder cancers. 73 74 75 76 77 There are now studies that demonstrate its possible use in HCC. 78 One of the earliest studies exploring the use of CTCs in HCC included 81 patients with HCC undergoing locoregional therapy.…”

Current State of Personalized Genitourinary Cancer Radiotherapy in the Era of Precision Medicine

“…Liquid biopsy tools such as CTCs and ctDNA are similarly being investigated as prognostic biomarkers in bladder cancer as they are in prostate cancer (115,116). CTCs and ctDNA will be collected and assessed in both the abovementioned INTACT and INSPIRE trials to determine their role as predictive biomarkers for overall outcomes after bladder preservation therapy.…”

“…The presence of circulating biomarkers is also being explored in the surveillance setting post-treatment. One study in patients undergoing neoadjuvant chemotherapy followed by radical cystectomy found that the presence of ctDNA was prognostic for worse outcomes overall (115). This has not yet been evaluated in a TMT cohort but the data from INTACT and INSPIRE will help elucidate the role of liquid biomarkers in TMTspecific cohorts.…”