Prognostic value of preoperative circulating tumor cells counts in patients with UICC stage I-IV colorectal cancer

The isolation of circulating tumour cells (CTCs) in colorectal cancer (CRC) mostly relies on the expression of epithelial markers such as EpCAM, and phenotypic characterisation is usually performed under fluorescence microscopy with only one or two additional markers. This limits the ability to detect different CTC subpopulations based on multiple markers. The aim of this work was to develop a novel protocol combining two platforms (IsoFluxTM and ImageStream®X) to improve CTC evaluation. Cancer cell lines and peripheral blood from healthy donors were used to evaluate the efficiency of each platform independently and in combination. Peripheral blood was extracted from 16 early CRC patients (before loco-regional surgery) to demonstrate the suitability of the protocol for CTC assessment. Additionally, peripheral blood was extracted from nine patients one month after surgery to validate the utility of our protocol for identifying CTC subpopulation changes over time. Results: Our protocol had a mean recovery efficiency of 69.5% and a limit of detection of at least four cells per millilitre. We developed an analysis method to reduce noise from magnetic beads used for CTC isolation. CTCs were isolated from CRC patients with a median of 37 CTCs (IQ 13.0–85.5) at baseline. CTCs from CRC patients were significantly (p < 0.0001) larger than cytokeratin (CK)-negative cells, and patients were stratified into two groups based on BRAFV600E and PD-L1 expression on CK-positive cells. The changes observed over time included not only the number of CTCs but also their distribution into four different subpopulations defined according to BRAFV600E and PD-L1 positivity. We developed a novel protocol for semi-automatic CTC isolation and phenotypic characterisation by combining two platforms. Assessment of CTCs from early CRC patients using our protocol allowed the identification of two clusters of patients with changing phenotypes over time.

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Deep Phenotypic Characterisation of CTCs by Combination of Microfluidic Isolation (IsoFlux) and Imaging Flow Cytometry (ImageStream)

Ruiz-Rodríguez

Molina-Vallejo

Aznar-Peralta

et al. 2021

“…In the last decades, biomarkers for micrometastasis like circulating and disseminating tumor cells (CTC and DTC) were identified. Tumor dissemination in peripheral blood and bone marrow proved to be associated with shorter OS and PFS in different solid tumors and PDAC [ 4 , 5 , 18 ]. DTCs stay dormant in the bone marrow and recirculate into various sites causing metastasis or even tumor recurrence [ 19 ].…”

Section: Discussionmentioning

confidence: 99%

Evaluation of the Hamburg-Glasgow Classification in Pancreatic Cancer: Preoperative Staging by Combining Disseminated Tumor Load and Systemic Inflammation

Abdalla

Almanfalouti

Effenberger

et al. 2021

Self Cite

This study aims to compare the Hamburg Glasgow Classification (HGC) to Union for International Cancer Control (UICC) classification in patients with pancreatic ductal adenocarcinoma (PDAC). As adequate tumor classification is only possible after tumor resection and histological evaluation, only 20% of patients with PDAC receive accurate tumor staging. Thus, an accurate preoperative staging system is still missing but urgently needed. Systemic inflammation and tumor dissemination are important factors regarding the oncological outcome. HGC integrates both into a preoperative staging system, by combining C-reactive protein (CRP), albumin, and disseminated tumor cells (DTC) in the bone marrow. In this prospective study, 109 patients underwent surgical exploration for suspected PDAC. All patients underwent a preoperative bone marrow aspiration for DTC detection. HGC showed significant preoperative risk stratification for overall survival (OS) (p-value < 0.001) and progression-free survival (PFS) (p-value < 0.001). These results were comparable to the UICC survival stratification for OS and PFS (p-value = 0.001 and 0.006). Additionally, in non-metastatic PDAC, HGC III-IV was associated with shorter OS and PFS (p-value < 0.001, respectively) when compared to HGC I-II. Therefore, the HGC is a promising preoperative prognostic staging classification for accurate and simple outcome stratification in patients with PDAC.

“…Plasma DNA can be analyzed by approaches targeting specific tumor-associated genes (e.g., mutations in the EGFR gene in non-small cell lung carcinoma (NSCLC)) or non-targeted screening approaches such as array CGH, whole-genome sequencing, or exome sequencing) [ 25 , 26 ]. In general, targeted approaches have higher analytical sensitivity than non-targeted approaches, but there are strong efforts to improve the detection limits of non-targeted approaches [ 27 , 28 ].…”

Section: Technologies For Ctc and Ctdna Analysesmentioning

confidence: 99%

“…According to Bork et al, the detection of CTCs in nodal negative CRC (UICC stage I–II) is as low as 9% [ 34 , 35 ]. In another study, the detection of CTCs using Cellsearch ® in CRC (UICC stage I–IV) was 45% [ 26 ]. Therefore, the utility of CTC-based screening using Cellsearch ® is still rather challenging and still not applicable ( Table 1 ).…”

Section: Clinical Applications Of Circulating Tumor Cells (Ctcs)mentioning

confidence: 99%

Clinical Applications of Circulating Tumor Cells and Circulating Tumor DNA as a Liquid Biopsy Marker in Colorectal Cancer

Heidrich

Abdalla

Reeh

et al. 2021

Self Cite

Colorectal cancer (CRC) is the third most commonly diagnosed cancer worldwide. It is a heterogeneous tumor with a wide genomic instability, leading to tumor recurrence, distant metastasis, and therapy resistance. Therefore, adjunct non-invasive tools are urgently needed to help the current classical staging systems for more accurate prognostication and guiding personalized therapy. In recent decades, there has been an increasing interest in the diagnostic, prognostic, and predictive value of circulating cancer-derived material in CRC. Liquid biopsies provide direct non-invasive access to tumor material, which is shed into the circulation; this enables the analysis of circulating tumor cells (CTC) and genomic components such as circulating free DNA (cfDNA), which could provide the key for personalized therapy. Liquid biopsy (LB) allows for the identification of patients with a high risk for disease progression after curative surgery, as well as longitudinal monitoring for disease progression and therapy response. Here, we will review the most recent studies on CRC, demonstrating the clinical potential and utility of CTCs and ctDNA. We will discuss some of the advantages and limitations of LBs and the future perspectives in the field of CRC management.