A Liquid Biopsy Assay for Noninvasive Identification of Lymph Node Metastases in T1 Colorectal Cancer

Abstract

BACKGROUND & AIMS:We recently reported use of tissuebased transcriptomic biomarkers (microRNA [miRNA] or messenger RNA [mRNA]) for identification of lymph node metastasis (LNM) in patients with invasive submucosal colorectal cancers (T1 CRC). In this study, we translated our tissuebased biomarkers into a blood-based liquid biopsy assay for noninvasive detection of LNM in patients with high-risk T1 CRC. METHODS: We analyzed 330 specimens from patients with high-risk T1 CRC, which included 188 serum samples from 2 clinical cohorts-a training cohort (N ¼ 46) and a validation cohort (N ¼ 142)-and matched formalin-fixed paraffinembedded samples (N ¼ 142). We performed quantitative reverse-transcription polymerase chain reaction, followed by logistic regression analysis, to develop an integrated transcriptomic panel and establish a risk-stratification model combined with clinical risk factors. RESULTS: We used comprehensive expression profiling of a training cohort of LNM-positive and LMN-negative serum specimens to identify an optimized transcriptomic panel of 4 miRNAs (miR-181b, miR-193b, miR-195, and miR-411) and 5 mRNAs (AMT, forkhead box A1 [FOXA1], polymeric immunoglobulin receptor [PIGR], matrix metalloproteinase 1 [MMP1], and matrix metalloproteinase 9 [MMP9]), which robustly identified patients with LNM (area under the curve [AUC], 0.86; 95% confidence interval [CI], 0.72-0.94). We validated panel performance in an independent validation cohort (AUC, 0.82; 95% CI, 0.74-0.88).Our risk-stratification model was more accurate than the panel and an independent predictor for identification of LNM (AUC, 0.90; univariate: odds ratio [OR], 37.17; 95% CI,; P < .001; multivariate: OR, 17.28; 95% CI, 1.82-164.07; P ¼ .013). The model limited potential overtreatment to only 18% of all patients, which is dramatically superior to pathologic features that are currently used (92%). CONCLUSIONS: A novel risk-stratification model for noninvasive identification of T1 CRC has the potential to avoid unnecessary operations for patients classified as high-risk by conventional risk-classification criteria.

DENEB: Development of new criteria for curability after local excision of pathological T1 colorectal cancer using liquid biopsy

“…However, LNM has only been identified in about 10% of cases, and the assessment based on the established pathological risk factors is limited due to high interobserver variability between pathologists 8 . More recently, the effectiveness of artificial intelligence using clinicopathological factors and the transcriptomic panel based on microRNAs and messenger RNAs in the blood for identification of LNM have been explored; however, none of these methodologies are well established and have sufficient evidence 9,10 . To our knowledge, this is the first study to explore the predictability of LNM using the established ctDNA assay from a different perspective than the pathological standpoint, in which prediction of LNM has been discussed for many years.…”

“…However, LNM has only been identified in about 10% of cases, and the assessment based on the established pathological risk factors is limited due to high interobserver variability between pathologists 8. More recently, the effectiveness of artificial intelligence using clinicopathological factors and the transcriptomic panel based on microRNAs and messenger RNAs in the blood for identification of LNM have been explored; however, none of these methodologies are well established and have sufficient evidence 9,10. To our knowledge, this is the first study to explore the predictability of LNM using the established ctDNA assay from a different perspective than the pathological standpoint, in which prediction of LNM has been discussed for many years.It will be possible to follow up pT1 patients with negative ctDNA after complete local resection and to evaluate the prognosis, provided a sufficient diagnostic concordance rate can be obtained.Reducing unnecessary surgery could contribute to the avoidance of postsurgical complications and preservation of patients' quality of life by preventing deterioration of their defecation function and avoiding stoma in cases of rectal cancer.…”

Blood-Based Liquid Biopsies: A Noninvasive and Cost-Effective Tool for Improved Risk Assessment and Identification of Lymph Node Metastasis in Patients With Submucosal T1 Colorectal Cancer

“…Many biomarker panels have gone through such rigorous interrogations (eg, ColoGuard), which led to approvals by the US Food and Drug Administration for specific clinical applications. 8 In this issue of the Gastroenterology, Wada et al, 9 describe the process through which they developed a transcriptomic signature that consisted of both mRNA and microRNAs (miRNAs) for the identification of patients with T1 CRCs and presence of lymph nodes metastasis (LNM). This has potential eventual clinical application for helping to reduce overtreatment in those who are deemed to be lymph node negative.…”

“…Overall, Wada et al, 9 made a case for a transcriptomic liquid biopsy assay to tease apart LNM-positive from LNM-negative T1 CRCs. Such biomarker panels were also proposed to assess the emergence of therapy resistance, residual disease, and recurrence.…”

Noninvasive Identification of Lymph Node Metastasis in T1 Colorectal Cancer by Transcriptomic Panel Based on Liquid Biopsy Assay: Additional Analytical Strategies Are Needed

“…We read with great interest the article entitled "A liquid biopsy assay for noninvasive identification of lymph node metastases in T1 colorectal cancer" by Wada et al 1 In that study, the authors detected their previously reported transcriptomic biomarkers (miRNAs and mRNAs) with the use of liquid biopsy methods and evaluated the predictive value of this transcriptomic panel for the noninvasive identification of lymph node metastasis (LNM) in patients with high-risk stage T1 colorectal cancer (CRC). The authors successfully established a new risk model of transcriptome profiles based on blood liquid biopsy assay that can reliably predict LNM in patients with stage T1 CRC.…”