Circulating Tumor Cell Kinetics and Morphology from the Liquid Biopsy Predict Disease Progression in Patients with Metastatic Colorectal Cancer Following Resection

Abstract

The liquid biopsy has the potential to improve current clinical practice in oncology by providing real-time personalized information about a patient’s disease status and response to treatment. In this study, we evaluated 161 peripheral blood (PB) samples that were collected around surgical resection from 47 metastatic colorectal cancer (mCRC) patients using the High-Definition Single Cell Assay (HDSCA) workflow. In conjunction with the standard circulating tumor cell (CTC) enumeration, cellular morphology and kinetics between time-points of collection were considered in the survival analysis. CTCs, CTC-Apoptotic, and CTC clusters were found to indicate poor survival with an increase in cell count from pre-resection to post-resection. This study demonstrates that CTC subcategorization based on morphological differences leads to nuanced results between the subtypes, emphasizing the heterogeneity within the CTC classification. Furthermore, we show that factoring in the time-point of each blood collection is critical, both for its static enumeration and for the change in cell populations between draws. By integrating morphology and time-based analysis alongside standard CTC enumeration, liquid biopsy platforms can provide greater insight into the pathophysiology of mCRC by highlighting the complexity of the disease across a patient’s treatment.

Characterization of BCMA Expression in Circulating Rare Single Cells of Patients with Plasma Cell Neoplasms

“…Image generation, technical analysis, and rare cell detection on the slide-based HDSCA immunofluorescence technology have been described previously [ 16 , 18 , 20 , 34 ]. Briefly, stained slides are imaged at 100× magnification, and an unsupervised clustering algorithm identifies rare cells based on cellular morphology and marker expression across all 2–3 million cells per slide, as described by 761 features per cell.…”

“…In this study, we report the technical development and initial genomic and clinical validation of a new slide-based, enrichment-free immunofluorescence assay (i.e., BCMA, CD138, CD45, DAPI) for the detection of circulating rare cells and morphogenomic profiling of BCMA+ cells in PC malignancies, hereafter referred to as the “BCMA assay”. The technical methodology is built on the established “no-cell-left-behind” approach of the high-definition single-cell assay (HDSCA) workflow—previously validated clinically for various pathologies, including breast cancer [ 14 ], myocardial infarction [ 15 ], melanoma [ 15 ], prostate cancer [ 16 ], bladder cancer [ 17 ], colorectal cancer [ 18 ], and multiple myeloma [ 19 , 20 ]—and has been optimized for the detection of BCMA expression for the characterization of MM precursor cells in PC neoplasia.…”

Defining A Liquid Biopsy Profile of Circulating Tumor Cells and Oncosomes in Metastatic Colorectal Cancer for Clinical Utility

“…Previous investigations have correlated CTC counts to CEA levels, with the two used in conjunction to accurately predict survival outcomes [13]. Generally, a higher number of CTCs indicates poorer patient outcomes in CRC [14,15], although nuances emerge when considering morphologically defined CTC subtypes and the change in cell populations over time [16,17]. With the variety of detectable biomarkers in circulation, the liquid biopsy could aid in tackling some of the clinical challenges of CRC.…”

“…However, CellSearch ® and similar systems are not commonly utilized by clinicians treating CRC [23,24]. This limited utility could be attributed to a lack of timepoint-specific standards [18] and infrequent CTC kinetics analysis, despite its clinical promise [16]. Most early-generation liquid-biopsy platforms employ enrichment-based approaches that detect a limited CTC population in the peripheral blood (PB) of CRC patients [25][26][27] and overlook other cellular subtypes with nuanced survival implications [16,28].…”

“…This limited utility could be attributed to a lack of timepoint-specific standards [18] and infrequent CTC kinetics analysis, despite its clinical promise [16]. Most early-generation liquid-biopsy platforms employ enrichment-based approaches that detect a limited CTC population in the peripheral blood (PB) of CRC patients [25][26][27] and overlook other cellular subtypes with nuanced survival implications [16,28]. In a cancer type known for its tumor heterogeneity like CRC, enrichment-based approaches limit the liquid biopsy's potential clinical utility, thereby warranting an unbiased single-cell approach that focuses on all rarity in the bloodstream.…”