Circulating Tumor Markers Research Articles

B-149 Diverse glycosylation of circulating alpha fetoprotein in chronic liver diseases profiled by intact mass high resolution mass spectrometry

Abstract Background Alpha fetoprotein (AFP) is a circulating tumor marker utilized clinically to detect liver cirrhosis, liver cancer and testicular seminomas. AFP molecule contains one single N-glycosylation site, with highly variable glycan structures, resulting in diverse AFP glycoforms in circulating blood. The core fucosylation of AFP (AFP-L3) is a clinical biomarker to monitor and predict risks of liver cancer progression and recurrence. Clinical testing for AFP-L3 utilizes lectin-binding and gel-shift electrophoresis. We developed a novel method to measure AFP glycoforms, based on immune-enrichment and liquid chromatography with high resolution mass spectrometry (LC-HRMS). Methods Immune enrichment of AFP was achieved by anti-AFP antibody, immobilized to magnetic beads. After binding and enrichment, the magnetic beads were washed, AFP was eluted, and the samples were analyzed by LC-HRMS (Exploris 480 MS and Vanquish UHPLC, Thermo Fisher Scientific), using full scan, positive polarity acquisition mode. Chromatographic separation was performed on PLRP column (Agilent); flow rate 200 µL/min. The HRMS data were processed and deconvoluted with intact mass analysis workflow in Biopharma Finder 5.1 (Thermo Fisher Scientific) to derive the intact AFP masses. The method’s sensitivity and imprecision were assessed by replicate analysis of serial diluted patient serum samples. AFP and its associated glycoforms were matched and identified with an in-house developed bioinformatics workflow. The deconvoluted intact masses and the sum intensity of detected charged species for each AFP glycoform were used to quantify relative abundance of AFP glycoforms. Results The method specificity for AFP was confirmed by bottom-up analysis after immune-enrichment with observed peptide sequence coverage over 35%. The recovery of the immune enrichment was shown to be independent of the AFP glycoform compositions. The identified AFP glycoforms and their relative abundance were consistent among replicates between AFP concentration 100 ng/mL and 5000 ng/mL. Analysis of authentic serum specimens from patients with different chronic liver diseases and liver cancer (n=102), revealed multiple complex biantennary branched AFP glycan modifications, with different numbers of terminal sialic acid and core fucose. Four AFP glycoforms were identified in over 60% patient samples with elevated AFP concentrations. Although occurring consistently, relative abundance of these AFP glycoforms varied significantly among patients. Assessment of the AFP fucosylation and sialylation features suggested that the two are orthogonal processes. Clinical correlations suggested that fewer terminal sialic acids were associated with impaired liver functions, while the fucosylation was linked to inflammatory responses and presence of liver malignancy. Strong linear correlation (Spearman’s rs=0.952, N=35) was observed between AFP-L3% quantitated by the LC-HRMS method, and AFP-L3% determined by the lectin-binding based isotachophoresis assay (Wako Diagnostics). Discrepancies between the two methods were observed in specimens containing < 400 ng/mL of AFP, in which the LC-HRMS method detected only a single predominant glycoform. Conclusions We developed and evaluated the LC-HRMS method for profiling circulating AFP glycoforms in patients with chronic liver diseases and liver cancer. The detected AFP glycoforms and their relative abundance were used to establish glycan features, which can be further linked to cancer development, impaired liver function, and inflammation.

Pre-analytical stability of the CEA, CYFRA 21.1, NSE, CA125 and HE4 tumor markers.

For lung cancer, circulating tumor markers (TM) are available to guide clinical treatment decisions. To ensure adequate accuracy, pre-analytical instabilities need to be known and addressed in the pre-analytical laboratory protocols. This study investigates the pre-analytical stability of CA125, CEA, CYFRA 21.1, HE4 and NSE for the following pre-analytical variables and procedures; i) whole blood stability, ii) serum freeze-thaw cycles, iii) electric vibration mixing and iv) serum storage at different temperatures. Left-over patient samples were used and for every investigated variable six patient samples were used and analysed in duplicate. Acceptance criteria were based on analytical performance specifications based on biological variation and significant differences with baseline. Whole blood was stable for at least 6 hours for all TM except for NSE. Two freeze-thaw cycles were acceptable for all TM except CYFRA 21.1. Electric vibration mixing was allowed for all TM except for CYFRA 21.1. Serum stability at 4°C was 7 days for CEA, CA125, CYFRA 21.1 and HE4 and 4 hours for NSE. Critical pre-analytical processing step conditions were identified that, if not taken into account, will result in reporting of erroneous TM results.

The diagnostic potential of miR-196a-1 in colorectal cancer

BackgroundColorectal cancer (CRC) is a common malignancy worldwide. MicroRNAs (miRNAs) are important epigenetic alterations that notably impact various physiological and pathological processes by acting as negative regulators of gene expression. Furthermore, they have a vital function in different types of cancers, including CRC. In this research, we evaluated, for the very first time, the expression levels of miR-196a-1 in the tissue and plasma of patients with CRC and also homeobox D8 (HOXD8) as the target gene.Materials and methodsThis study included a collection of 220 plasma and tissue samples from 55 patients diagnosed with CRC, as well as 55 healthy individuals matched by age and sex. Total RNA was extracted from plasma and tissue samples, and then polyadenylation and cDNA synthesis were performed. The expression levels of miR-196a-1 and HOXD8 as target gene was evaluated by quantitative RT-PCR (qRT-PCR) assay. We compared the diagnostic value of plasma miR-196a-1 with that of the circulating tumor markers CA19-9 and CEA using a Receiver Operating Characteristics (ROC) analysis. The association of miR-196a-1 with clinicopathological characteristics was assessed in tissue and plasma samples from patients with CRC.ResultsOur data demonstrated that the expression levels of miR-196a-1 in the tissue and plasma samples of CRC patients were 11.426- and 11.655-fold higher, respectively than those in adjacent normal tissue and plasma samples from normal subjects (p < 0.001). Through ROC curve analysis, it was identified that the sensitivity and specificity of miR-196a-1 for tissue samples, with an AUC of 0.925, were 89% and 98%, respectively. In addition, the sensitivity and specificity for plasma samples with an AUC of 0.801 were 70% and 98%, respectively. These findings reveal that miR-196a-1 is a useful biomarker for discriminating cases from controls. Furthermore, the expression of HOXD8 was not significantly altered in tumor tissue samples compared to adjacent normal tissues (P > 0.05).ConclusionsThese results show that miR-196a-1 has an oncogenic impact and plays a significant role in CRC development. The results also indicate that miR-196a-1 could serve as a novel noninvasive biomarker for the detection of CRC.

Long Noncoding RNAs MALAT1 and HOTTIP Act as Serum Biomarkers for Hepatocellular Carcinoma.

Circulating tumor markers with satisfactory sensitivity and specificity play crucial roles in cancer diagnosis and therapy. This prospective study aimed to evaluate the potential of circulating lncRNAs as biomarkers for hepatocellular carcinoma (HCC). A total of 74 patients with HCC and 94 healthy controls were enrolled. The expression levels of candidate genes in serum were detected by qRT-PCR. Receiver operating characteristic (ROC) curve analysis and logistic regression were employed to investigate the diagnostic capacity of lncRNAs. The analysis of 3-year overall survival (OS) was conducted using the Kaplan-Meier method and log-rank test. Of the 9 candidate genes, 6 lncRNAs could be stably detected in serum. The expression levels of circulating MALAT1 and HOTTIP in HCC patients were significantly higher than those in controls (P < 0.001). ROC analysis showed that MALAT1 and HOTTIP were more effective than alpha-fetoprotein (AFP) (P < 0.010) in the diagnosis of HCC, with AUCs of 0.896 and 0.899, respectively. Additionally, a panel consisting of MALAT1, HOTTIP, and AFP was constructed to obtain an AUC of 0.968 with a sensitivity of 87.8% and specificity of 94.7% in HCC diagnosis. Moreover, the upregulation of MALAT1 was not only related to multiple tumor lesions, HCV infection, AST level, and AFP level, but also suggested shorter OS. A high expression level of HOTTIP was associated with metastasis. Serum MALAT1 and HOTTIP play indicative roles as non-invasive biomarkers for HCC.

DNA nanotechnology in tumor liquid biopsy: Enrichment and determination of circulating biomarkers

AbstractMonitoring tumor biomarkers in a non‐invasive manner for tumor diagnosis has attracted increasing attention. Liquid biopsy mainly includes three types of biomarkers: circulating tumor cells, extracellular vesicles, and circulating nucleic acids. These biomarkers released by tumor cells can travel to other parts of the circulation system. The analysis of circulating tumor markers in the circulation enables early tumor diagnosis, progression evolution, and treatment monitoring, which are important for individualized clinical decisions. In this study, we summarize different DNA nanotechnology strategies that can be used to capture, amplify, and measure circulating biomarkers and discuss the current challenges and outlook.

An Analysis of Patients Treated with Stereotactic Body Radiotherapy for Metastatic Urinary Tract Tumors to Identify Predictors of Response.

To identify selection criteria linked to outcomes in patients treated with stereotactic body radiotherapy (SBRT) for metastatic tumors of the urinary tract (UT). Single institution retrospective analysis of SBRT treated patients for oligometastatic/progressive UT tumors from 2006-2022. Charts were queried for M1 status at diagnosis or during disease course, treatment details (surgery, SBRT, systemic therapy), metabolic status (diabetes [DM], BMI) and outcomes. A linear quadratic formula was used to calculate the biologically effective dose (BED) using an α/β of 10 for tumor. Descriptive statistics portrayed the cohort, and analyses were done at patient and site level. Time-to-event analyses, including overall survival (OS) and progression-free survival (PFS) from SBRT, were assessed by the Kaplan-Meier method. Cox regression was used for univariable (UVA) and multivariable analyses (MVA) to identify predictors of outcomes. A total of 35 patients were treated at 44 metastatic sites, including: bone (25%), node (36.4%), lung (20.5%), soft tissue (13.6%) and liver (4.5%). Most were male (74.3%) with a median age of 70 (range: 51-89), without DM (60%) having a median BMI of 29.8, and ECOG <2 (97.1%) at time of SBRT. Six (17.1%) patients were M1 at diagnosis. Of the 29 non-M1 patients, 86.2% received definitive local therapy (LT), 58.6% had at least T3/N+ disease, 75.8% received systemic therapy with a median of 2 agents (range: 1-6) prior to SBRT. Sixteen (45.7%) received immunotherapy (IO) with most receiving this before (75%) and after (56.2%) SBRT. Six patients had positive PD-L1 status (n = 10). The median RT dose, fractionation and BED was 40 Gy (range: 14-46), 5 fractions, and 72 (range: 28-132), respectively. At a median follow-up of 34.8, the median OS was 18.4 m (range: 9.3-27.4) with a 2-year OS of 35.9%. At patient level, 62.8% recurred after SBRT. The median PFS after SBRT was 5.3 m (range: 1.8-8.7) with a 2-yr PFS of 29.3%. Patient-level PFS was improved with LT (6.7 vs 1.4 m; p = 0.001) and DM (NR vs 2.9 m; p = 0.015), whereas improved OS was related with LT (18.9 vs 6.6 m; p = 0.03), DM (p = 0.04), ECOG (p = 0.004), and no relapse after SBRT (NR vs 9.8 m; p <0.001). Exposure to < 3 systemic agents prior to SBRT portended better PFS (6.7 vs 2.6 m; p = 0.04) without any impact by IO. At site level, 20.4% of sites had local relapse with 4 being the first event. Site was related with PFS (p = 0.009) with order of increased relapse risk being liver > bone > soft tissue > node > lung. No dosimetric feature was related with recurrence risk. On MVA, both DM (p = 0.02) and LT (p = 0.002) were predictive for PFS. Only recurrence after SBRT predicted for OS on MVA (HR: 6.7, 95% CI: 1.4-31; p = 0.014). In the IO subset, median PFS was 5.3 m and OS was 9.4 m, with no difference seen with IO-SBRT sequence or PDL1 status. Optimized selection criteria for metastasis-directed therapy in patients with UT tumors is unclear, notably with IO. Future studies may benefit by assessing circulating tumor markers prior to SBRT.

Abstract 4338: Response to neoadjuvant targeted therapy in operable head and neck cancer confers survival benefit

Abstract Purpose: Neoadjuvant targeted therapy provides a brief, preoperative ‘window of opportunity’ that can be exploited to individualize cancer care based on treatment response. We investigated whether response to neoadjuvant therapy during the preoperative window confers survival benefit in patients with operable head and neck squamous cell carcinoma (HNSCC). Patients and Methods: A pooled analysis of treatment naïve patients with operable HNSCC enrolled in one of three clinical trials from 2009-2020 (NCT00779389, NCT01218048, NCT02473731). Neoadjuvant regimens consisted of EGFR inhibitors (n=83) or anti-ErbB3 antibody therapy (n=9) within 28 days of surgery. Clinical to pathologic stage migration was compared to disease-free survival (DFS) and overall survival (OS) while adjusting for confounding factors using multivariable Cox regression. Circulating tumor markers validated in other solid tumor models were analyzed. Results: 92 of 118 patients were analyzed; all patients underwent surgery following neoadjuvant therapy. Clinical to pathologic downstaging was more frequent in patients undergoing neoadjuvant targeted therapy compared to a historic cohort without neoadjuvant therapy (P=0.048). Patients with pathologic downstage migration had the highest OS (89.5%, 95% CI 75.7-100) compared to those with no stage change (58%, 95% CI 46.2-69.8) or upstage (40%, 95% CI 9.6-70.4, P=0.003). On multivariable analysis, downstage migration remained a positive prognostic factor for OS (hazard ratio 0.22, 95% CI 0.05-0.90) while adjusting for measured confounders. Downstage migration correlated with decreased circulating tumor markers, SOX17 and TAC1 (P=0.0078). Conclusion: Brief neoadjuvant therapy achieved pathologic downstaging in a subset of patients and was associated with significantly better DFS and OS as well as decreased circulating methylated SOX17 and TAC1. Citation Format: Marco Antonio Mascarella, Tolani Olonisakin, Purva Rumde, Varun Vendra, Melonie Nance, Seungwon Kim, Mark Kubik, Shaum Sridharan, Robert Ferris, Moon Fenton, Daniel Clayburgh, James Ohr, Malabika Sen, Sonali Joyce, James Herman, Jennifer Grandis, Dan Zandberg, Umamaheswar Duvvuri. Response to neoadjuvant targeted therapy in operable head and neck cancer confers survival benefit. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4338.

Myo1e overexpression in lung adenocarcinoma is associated with increased risk of mortality

This study aims to perform a comprehensive genomic analysis to assess the influence of overexpression of MYO1E in non-small cell lung carcinoma (NSCLC) and whether there are differences in survival and mortality risk in NSCLC patients depending on both DNA methylation and RNA expression of MYO1E. The DNA methylation probe cg13887966 was inversely correlated with MYO1E RNA expression in both LUAD and LUSC subpopulations showing that lower MYO1E RNA expression was associated with higher MYO1E DNA methylation. Late stages of lung cancer showed significantly lower MYO1E DNA methylation and significantly higher MYO1E RNA expression for LUAD but not for LUSC. Low DNA methylation as well as high RNA expression of MYO1E are associated with a shorter median survival time and an increased risk of mortality for LUAD, but not for LUSC. This study suggests that changes in MYO1E methylation and expression in LUAD patients may have an essential role in lung cancer’s pathogenesis. It shows the utility of MYO1E DNA methylation and RNA expression in predicting survival for LUAD patients. Also, given the low normal expression of MYO1E in blood cells MYO1E DNA methylation has the potential to be used as circulating tumor marker in liquid biopsies.

Response to Neoadjuvant Targeted Therapy in Operable Head and Neck Cancer Confers Survival Benefit.

Neoadjuvant targeted therapy provides a brief, preoperative window of opportunity that can be exploited to individualize cancer care based on treatment response. We investigated whether response to neoadjuvant therapy during the preoperative window confers survival benefit in patients with operable head and neck squamous cell carcinoma (HNSCC). A pooled analysis of treatment-naïve patients with operable HNSCC enrolled in one of three clinical trials from 2009 to 2020 (NCT00779389, NCT01218048, NCT02473731). Neoadjuvant regimens consisted of EGFR inhibitors (n = 83) or anti-ErbB3 antibody therapy (n = 9) within 28 days of surgery. Clinical to pathologic stage migration was compared with disease-free survival (DFS) and overall survival (OS) while adjusting for confounding factors using multivariable Cox regression. Circulating tumor markers validated in other solid tumor models were analyzed. 92 of 118 patients were analyzed; all patients underwent surgery following neoadjuvant therapy. Clinical to pathologic downstaging was more frequent in patients undergoing neoadjuvant targeted therapy compared with control cohort (P = 0.048). Patients with pathologic downstage migration had the highest OS [89.5%; 95% confidence interval (CI), 75.7-100] compared with those with no stage change (58%; 95% CI, 46.2-69.8) or upstage (40%; 95% CI, 9.6-70.4; P = 0.003). Downstage migration remained a positive prognostic factor for OS (HR, 0.22; 95% CI, 0.05-0.90) while adjusting for measured confounders. Downstage migration correlated with decreased circulating tumor markers, SOX17 and TAC1 (P = 0.0078). Brief neoadjuvant therapy achieved pathologic downstaging in a subset of patients and was associated with significantly better DFS and OS as well as decreased circulating methylated SOX17 and TAC1.

Exosomal hsa-piR1089 promotes proliferation and migration in neuroblastoma via targeting KEAP1.

Neuroblastoma (NB) is a sympathetic nervous system tumor and one of the most common pediatric, extra-cranial, solid tumors, especially in early childhood. Its expression is heterogeneous and shows a unique clinical and prognostic feature. Due to its insidious onset, most diagnoses are accompanied by metastasis, making patient prognoses extremely poor. Novel biomarkers are urgently needed for easy diagnosis, metastasis detection, and investigation of potential mechanisms regulating NB tumor progression. Recent research highlights that circulating tumor markers could be used to diagnose and monitor prognosis in various tumors. Among them, exosomal genetic material has attracted much attention because of its tumor-secreted specificity and unique mechanism of action. In this study, we used next-generation sequencing to study PIWI-interacting RNAs (piRNAs) in exosomes derived from NB patient plasma. We found higher human piRNA 1089 (hsa-piR-1089) levels in exosomes from NB patients than from normal controls. Our receiver operating characteristic (ROC) curve analyses showed that hsa-piR-1089 had high diagnostic sensitivity and specificity. We also found that high hsa-piR-1089 expression in NB tumor tissues was associated with a high-risk Children's Oncology Group classification and metastasis. Our in vitro experiments showed that exosomal hsa-piR-1089 promoted NB cell proliferation and migration by inhibiting Kelch-like ECH-associated protein 1 (KEAP1) expression. Moreover, low KEAP1 expression was associated with NB progression in clinical samples. In conclusion, our data indicate that blood-borne exosomal hsa-piR-1089 is a diagnostic marker for NB and assessing metastasis. Our study provides a quick, simple, and noninvasive diagnostic method for NB and contributes to developing new treatment strategies.

TUMOR MARKERS EXPRESSION LEVELS IN GASTRIC CANCER PATIENT'S PERIPHERAL BLOOD BY RT-PCR ASSESSMENT.

Hematological recurrence is the second most frequent cause of failure in the treatment of gastric cancer. The detection of circulating tumor markers in peripheral blood by quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) method may be a useful tool to predict recurrence and determine the patient's prognosis. However, no consensus has been reached regarding the association between the tumor markers level in peripheral blood and its impact on patient survival. To evaluate the expression of the circulating tumor markers CK20 and MUC1 in peripheral blood samples from patients with gastric cancer by qRT-PCR, and to verify the association of their expression levels with clinicopathological characteristics and survival. A total of 31 patients with gastric adenocarcinoma were prospectively included in this study. CK20 and MUC1 expression levels were analyzed from peripheral blood by the qRT-PCR technique. There was no statistically significant (p>0.05) association between CK20 expression levels and clinical, pathological, and surgical features. Higher MUC1 expression levels were associated with female patients (p=0.01). There was a correlation between both gene levels (R=0.81, p<0.001), and CK20 level and tumor size (R=0.39, p=0.034). CK20 and MUC1 expression levels could be assessed by qRT-PCR from total peripheral blood samples of patients with gastric cancer. CK20 levels were correlated to MUC1 levels as well as to tumor size. There was no difference in disease-free survival and overall survival regarding both genetic markers expression in this series.

Clinical usefulness of circulating tumor markers.

Tumor markers are a heterogeneous group of substances released by cancer cells into bloodstream, but also expressed by healthy tissues. Thus, very small concentrations can be present in plasma and serum from healthy subjects. Cancer patients tend to show increased levels correlating with tumor bulk, but false positive results could be present in patients with benign conditions. Thecorrect interpretation of TM results could be challenging and many factors should be considered, from pre-analytical conditions to patient concomitant diseases. In this line, the Clinical Chemistry and Laboratory Medicine journal has made important contributions though several publications promoting the adequate use of TM andtherefore improving patient safety. TM measurement offers valuable information for cancer patient management in different clinical contexts, such as helping diagnosis, estimating prognosis, facilitating early detection of relapse and monitoring therapy response. Our review analyzes the clinical usefulness of tumor markers applied in most frequent epithelial tumors, based on recent evidence and guidelines.

A novel combo of niraparib and anlotinib in platinum-resistant ovarian cancer, the final efficacy and safety report of ANNIE study, a phase II, multi-center trial (LBA 2)

<b>Objectives:</b> Patients with platinum-resistant ovarian cancer have a poor prognosis. Effective treatment options for these patients are limited. In this study (ANNIE), we evaluated the efficacy and safety of niraparib combined with anlotinib in patients with platinum-resistant recurrent (PRR) epithelial ovarian, fallopian tubal, or primary peritoneal cancer. <b>Methods:</b> The ANNIE trial (ClinicalTrials.gov identifier NCT04376073) was a multicentre, single-arm, phase 2 study. The patients with PRR and the measurable disease were given oral niraparib 300mg/200mg once daily continuously and anlotinib 12mg/10mg/8mg on day 1-14 of each 21-day cycle until disease progression or intolerable toxicity. The primary endpoint was the objective response rate (ORR) according to RECIST version 1.1. An exploratory analysis of archival peripheral blood was conducted in an attempt to analysis the correlation between circulating tumor markers and prognosis. <b>Results:</b> Between May 22, 2020, and April 22, 2021, 40 patients were finally enrolled. The subjects had previously received a median 4 lines (range, 1-9) of chemotherapy. Only 5 subjects had a deleterious germline <i>BRCA</i>1/2 mutation. At the cutoff date of analysis on December 31, 2021, 7 subjects were still on treatment. Thirty-six subjects underwent imaging evaluation. The confirmed best overall response showed 1 case with complete responses, 19 with partial responses, yielding the ORR of 55.6% (95% CI, 38.1%~ 72.1%). The median progression-free survival was 8.3 (95% CI, 5.86~ 10.81) months. The median overall survival was not reached. The most common drug-related adverse events were hypertension (55.0%), leucopenia (45.0%) and hand-foot syndrome (42.5%), and the most common Grade 3/4 events were neutropenia (17.5%), hypertension (12.5%) and anemia (12.5%). No treatment-related death was recorded. Preliminary analysis showed that patients with wild-type <i>PPM1D</i> had more significant changes in tumor size than those with <i>PPM1D</i> mutation (<i>P</i> =0.048). Further analysis of biomarkers and treatment prognostic results is ongoing and released soon. <b>Conclusions:</b> Niraparib in combination with anlotinib showed promising antitumor activity and tolerable toxicity in patients with platinum-resistant recurrent ovarian cancer.Fig. 1

P-158 Comprehensive recurrence risk assessment after colectomy, for stage II and III colorectal cancer patients, using genetic profiling, microbiome, and circulating tumor markers

Colorectal cancer (CRC) incidence increases in the western population and is considered of a major public health issue. To date, patients’ prognosis is mainly based on TNM staging of the primary tumor; however, clinical outcome varies among patients of same staging. Early diagnosis and treatment may lead to a better prognosis, especially when surgical excision and adjuvant treatment options are possible. Standard adjuvant treatment according to international guidelines is six months of Folfox or Capox.

Evaluation of Diagnostic and Prognostic Value of hsa_circ_0084927 and Analysis of Associated ceRNA Network in Colorectal Cancer.

ObjectThis study aims to analyze the differentially expressed circRNA in colon adenocarcinoma (COAD) and evaluate its diagnostic and prognostic value. Analyze associated circRNA-miRNA-mRNA network in COAD.Methods and MaterialsReal-time quantitative PCR (RT-PCR) was used to verify differentially expressed circRNA in COAD tissues and cells; Receiver operator characteristic (ROC) and Cox regression analysis were used to evaluating its diagnostic and prognostic value; Meanwhile we conducted CCK-8, invasion, and migration experiments in cell lines to explore the function of circRNA. In addition, a competitive endogenous RNA (ceRNA) network was established using bioinformatics methods to explore its prognostic value and potential functional mechanisms.ResultsOur study found that hsa_circ_0084927 is highly expressed in COAD tissues and cell lines. Plasma hsa_circ_0084927 can be used as a diagnostic marker for COAD patients; hsa_circ_0084927 can promote the proliferation, migration and invasion of COAD cells. In addition, we effectively constructed a ceRNA: network has_circ_0084927/miR-106b-5p/VEGFA. The ceRNA network indicates that hsa_circ_0084927 may affect the prognosis of COAD through the regulation of cell cycle, apoptosis and other pathways.ConclusionOur research results indicate that hsa_circ_0084927 has a cancer-promoting effect and may be used as a circulating tumor marker for COAD prognosis. In addition, this study proposes a new ceRNA network to provide new insights for the targeted therapy of COAD.

Appropriateness and Economic Analysis of Conventional Circulating Biomarkers Assessment in Early Breast Cancer: A Real-World Experience from the E.Pic.A Study.

The risk of relapse for early breast cancer (BC) patients persists even after decades and to date, no specific and sensitive effective circulating biomarker for recurrence prediction has been identified yet. The international guidelines do not recommend the assessment of the serum tumor markers CEA and CA15-3 in the follow-up of asymptomatic early BC patients. In our institute, IRCCS Istituto Romagnolo per lo Studio dei Tumori (IRST) “Dino Amadori”, as part of the E.Pic.A study, which was designed to assess the economic appropriateness of integrated care pathways in early BC, the use of CEA and CA15-3 as circulating tumor biomarkers in early BC patients was evaluated in 1502 patients one year after surgery, from 2015 to 2018, with an overall expense of EUR 51,764. A total of EUR 47,780 (92%) was used for execution of circulating tumor markers in early BC patients with stage 0, I and II tumors, neglecting the current guidelines and considered inappropriate by our professional board. We found that no patients with stage I BC experienced relapse in the 365 days after surgery, and in any case examination of the circulating markers CEA and CA15-3 was considered crucial for diagnosis of relapse. Our findings suggest that this inadequacy is a low-value area, supporting the reallocation of economic resources for interventions of a higher value for patients.

English

Biomarkers are considered as an innovative tool in the diagnosis and follow-up of breast cancer. The aim of the study was to assess the profile of circulating tumour markers CA 15-3 and CEA in patients under chemotherapy for breast cancer in Ouagadougou. This is a prospective cross-sectional study with a descriptive and analytical aims which was done from July to November 2020. Patients with histologically confirmed malignant breast tumour and under chemotherapy were included. Results revealed that the study was on thirty (30) female patients whose average age was 47.47 &plusmn; 2.10 years with a mean BMI of 27.29 &plusmn; 1.09 kg/m2. It was a non-specific type of infiltrating carcinoma with SBRm II grade in 90% of the patients. The mean CA 15-3 was 212.98 U/mL before chemotherapy and 165.75 U/mL after it. The CEA mean value was 3.13 ng/L before chemotherapy and 16.14 ng/L after it. Serum CA 15-3 was significantly associated with tumour site, SBRm grade, chemotherapy line and treatment response. Serum CEA level was significantly associated with tumour site and SBRm grade. Despite their lack of sensitivity, tumour markers, particularly CA 15-3 enabled assessment of the response to treatment in patients in this study. &nbsp; Key words: Tumour markers, CA 15-3, CEA, breast cancer, chemotherapy.

Metabolic changes in colorectal carcinomas are key factors for the early detection of neoplastic change

Potential metabolic biomarkers have been developed by the use of modern analytical techniques and nanotechnology in metabolomics, providing insight into the pathophysiological basis and changes, tumorigenesis, and molecular mechanisms that underpin better therapeutic, monitoring, and prognostic evaluations of colon malignancies. This would allow early detection and characterization of malignant colon tumors and could reduce the risk of mortality and morbidity of colorectal carcinomas. Based on their association with certain metabolic pathways linked to malignancies, a number of tumor markers have been designed. Whereas some have been associated with only one cancer type, while others are associated with many different forms of cancer. No tumor marker has been found to have universal application as a metabolism-related marker; although some are circulating tumor markers found in blood, urine, stool, or other body fluids, others may be found in the specific tumors themselves. This paper addresses a number of associated metabolic changes linked to colorectal cancers and potential applications for disease condition diagnosis, monitoring, treatment, and prognosis.

P-186 Vimentin-positive circulating tumor cells as diagnostic and prognostic biomarkers in biliary tract cancer patients

Biliary tract cancer (BTC) has poor prognosis and its mortality is higher than other gastrointestinal malignancies. Early diagnosis of BTC is important to decrease the mortality. Vimentin-positive circulating tumor cells (V-CTCs) are a good candidate for diagnostic biomarkers. However, there are limited studies regarding the role of V-CTCs as diagnostic and prognostic biomarkers in BTC patients. The aim of this study was to evaluate the diagnostic efficacy of V-CTCs between BTC and benign biliary disease (BBD) and determine the prognostic value of V-CTCs in BTC patients. We recruited 69 participants who has BTCs and BBDs from a single tertiary referral center between June 2018 and February 2021. We analyzed CTCs and V-CTCs using peripheral blood by CD-PRIMETM system, which is a commercialized version of the FAST disc (Clinomics, Ulsan, Korea). CTCs were detected in 62 of 69 (90%) patients. CTC count > 40/ml blood (20% Vs 55.8%, p=0.039), V-CTC > 15/ml blood (10% Vs 57.7%, p=0.005) and V-CTC/CTC ratio > 40% (10% Vs 48.1%, p=0.025) were significantly different between BTCs and BBDs. And two or more of these parameters (10% Vs 61.5%, p=0.002) increased the accuracy of distinguishing between BTCs and BBDs. Moreover, the combination of circulating tumor markers with elevated CA19-9 and biopsy, increased the accuracy (10% Vs 90.4%, p=0.000). Combination of circulating tumor markers with elevated CA19-9 (10%, Vs 84.8%, p=0.000) and biopsy (10% Vs 87.9%, p=0.000) was also distinguished between BBD and resectable BTC. In terms of prognosis, V-CTC > 50/ml blood was a significantly influencing factor of survival (140 days Vs 253 days, p=0.008). The combination of circulating tumor markers with elevated CA19-9 and/or biopsy may discriminate early stage of BTC from BBD, and V-CTC could determine future prognosis in patients with BTC.

The Clinical Significance of DJ1 and L1CAM Serum Level Monitoring in Patients with Endometrial Cancer.

Circulating tumor markers are not routinely used in patients with endometrial cancer (EC). This pilot study evaluated the role of monitoring new biomarkers DJ1 and L1CAM, in correlation with CA125 and HE4, for the effects of anticancer treatment and preoperative management in EC patients. Serial serum levels of DJ1, L1CAM, CA125 and HE4 were collected in 65 enrolled patients. Serum DJ1, L1CAM, CA125 and HE4 levels were significantly higher at the time of diagnosis compared to those measured during follow-up (FU). In patients with recurrent disease, serum DJ1, CA125 and HE4 levels were significantly higher at the time of recurrence compared to levels in disease-free patients. Serum L1CAM levels were also higher in patients with recurrence but without reaching statistical significance. While DJ1 levels were not affected by any of the observed patient-related characteristics, L1CAM levels were significantly higher in patients with age ≥60 years who were overweight. At the time of EC diagnosis, DJ1 and L1CAM serum levels did not correlate with stage, histological type or risk of recurrence. This is a preliminary description of the potential of serial DJ1 and L1CAM serum level measurement for monitoring the effects of treatment in EC patients.