Abstract LB-476: A universal method for elimination of haemolyzed plasma samples that improves miRNA signature performance for early detection of colorectal cancer

2012 ◽  
Author(s):  
Søren J. Nielsen ◽  
Thorarinn Blondal ◽  
Maria W. Teilum ◽  
Claus L. Andersen ◽  
Torben Orntoft ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Early Detection ◽  
Universal Method ◽  
Plasma Samples ◽  
Mirna Signature

Validation of a plasma-based miRNA PCR test for early detection of colorectal cancer.

2012 ◽  
Vol 30 (4_suppl) ◽  
pp. 424-424
Author(s):  
Søren Jensby Nielsen ◽  
Hanni Willenbrock ◽  
Jacob Fog ◽  
Jan Stenvang ◽  
Thorarinn Blondal ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Early Detection ◽  
Minimally Invasive ◽  
Melting Curve ◽  
Plasma Samples ◽  
Mirna Signature ◽  
Invasive Approach ◽  
Screening Programs ◽  
Screening Population ◽  
Plasma Mirna

424 Background: Colorectal cancer (CRC) is a major cause of mortality in the western world. Early detection of CRC improves survival and screening for CRC has been clinically proven to lower CRC-related mortality in the screening population. However, although population screening programs have been implemented in a number of countries, screening rates among the 50-75 year olds are unsatisfactory. There is therefore a clear unmet need for a quick, sensitive, specific, and minimally invasive screening assay to select at risk individuals for definitive diagnosis by colonoscopy. Methods: In order to detect microRNA (miRNA) biomarkers for CRC in blood plasma, we developed an LNA-enhanced miRNA RT-qPCR platform with high sensitivity and linearity for optimal quantitation of miRNAs from limited plasma samples. A clinical reference- lab compatible workflow that allows for the entire procedure from sample preparation through data acquisition and QC to test result to be completed within one working day was established. A reference melting curve database has been implemented to ensure the integrity of each data point, and appropriate controls monitor plate-to-plate and day-to-day variation. State-of-the-art normalization protocols have been evaluated to ensure optimal normalization of datasets prior to data analysis. Results: We previously determined a miRNA signature in a multi hospital discovery cohort that is differentially expressed between healthy individuals and stage II CRC patients. Here we report on the validation of this miRNA signature in an independent set of plasma samples from CRC patients and healthy volunteers. We have counter-screened the miRNA signature in a set of patients with other prevalent diseases, including hypertension, diabetes, diverticulitis, and others. Conclusions: A plasma miRNA signature for early detection of CRC from patient plasma was developed and validated in an independent clinical sample set. The signature was specific with respect to other diseases prevalent in the screening population. A second large-scale validation project is on-going. We conclude that plasma miRNA biomarkers can constitute an effective minimally invasive approach to population-wide CRC screening.

scholarly journals Gel-Based Proteomics of Clinical Samples Identifies Potential Serological Biomarkers for Early Detection of Colorectal Cancer

2019 ◽  
Vol 20 (23) ◽  
pp. 6082 ◽  
Author(s):  
Stine Thorsen ◽  
Irina Gromova ◽  
Ib Christensen ◽  
Simon Fredriksson ◽  
Claus Andersen ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Early Detection ◽  
Biomarker Discovery ◽  
Real Life ◽  
Clinical Samples ◽  
Plasma Samples ◽  
Healthy Individuals ◽  
Protein Biomarkers ◽  
2D Gel ◽  
Cancer Diseases

The burden of colorectal cancer (CRC) is considerable—approximately 1.8 million people are diagnosed each year with CRC and of these about half will succumb to the disease. In the case of CRC, there is strong evidence that an early diagnosis leads to a better prognosis, with metastatic CRC having a 5-year survival that is only slightly greater than 10% compared with up to 90% for stage I CRC. Clearly, biomarkers for the early detection of CRC would have a major clinical impact. We implemented a coherent gel-based proteomics biomarker discovery platform for the identification of clinically useful biomarkers for the early detection of CRC. Potential protein biomarkers were identified by a 2D gel-based analysis of a cohort composed of 128 CRC and site-matched normal tissue biopsies. Potential biomarkers were prioritized and assays to quantitatively measure plasma expression of the candidate biomarkers were developed. Those biomarkers that fulfilled the preset criteria for technical validity were validated in a case-control set of plasma samples, including 70 patients with CRC, adenomas, or non-cancer diseases and healthy individuals in each group. We identified 63 consistently upregulated polypeptides (factor of four-fold or more) in our proteomics analysis. We selected 10 out of these 63 upregulated polypeptides, and established assays to measure the concentration of each one of the ten biomarkers in plasma samples. Biomarker levels were analyzed in plasma samples from healthy individuals, individuals with adenomas, CRC patients, and patients with non-cancer diseases and we identified one protein, tropomyosin 3 (Tpm3) that could discriminate CRC at a significant level (p = 0.0146). Our results suggest that at least one of the identified proteins, Tpm3, could be used as a biomarker in the early detection of CRC, and further studies should provide unequivocal evidence for the real-life clinical validity and usefulness of Tpm3.

DNA methylation profiling from circulating tumor DNA for early-detection of colorectal cancer.

2019 ◽  
Vol 37 (15_suppl) ◽  
pp. e15076-e15076
Author(s):  
Wei Zhang ◽  
Jinke Sui ◽  
Xianrui Wu ◽  
Fuao Cao ◽  
Guanyu Yu ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Dna Methylation ◽  
Early Detection ◽  
Methylation Status ◽  
Stage I ◽  
Support Vector ◽  
Plasma Samples ◽  
Neoplastic Progression ◽  
Healthy Individuals ◽  
Early Stage Disease

e15076 Background: Colorectal cancer (CRC) develops as a result of neoplastic progression, which often takes decades, providing a window for early detection. Unfortunately, there has been little success in developing blood-based screening method due to the low amount of ctDNA present in the circulation, especially in patients with early stage disease. The role of aberrant DNA methylation, occurring very early in tumorigenesis, has been well elucidated. In this prospective study, we evaluated the potentiality of DNA methylation status obtained from ctDNA as an early detection method. Methods: Panel Design: Methylation data of tumor samples (12 types, n = 4,772), adjacent normal (8 types, n = 411), and normal white blood cells (n = 656) from TCGA and GSE were compared. Differentially methylated sites were extracted using modified wald-test with an adjusted p-value < 0.05 and fold-change > 2. Our panel covers 80,672 CpG sites, spanning 1.05Mb of human genome. We performed targeted bisulfite sequencing on plasma samples of 67 (stage I: 13, II:29, III: 23, IV: 2) Chinese CRC patients and 144 healthy individuals to construct a model for deriving markers that are differentially methylated and their associated weight. The model was validated in 2 independent cohorts. Results: We constructed a model using a support vector machine (SVM)-based machine learning classifier based on top 4,000 differentially methylated regions (DMRs) selected by random forest between tumor and normal plasma samples. Subsequently, 5-fold cross-validation with 100-time repeats were performed to gain a robust estimation of model performance, achieving a sensitivity of 91%, specificity of 98% and area under curve (AUC) of 98.6%. The model was subsequently validated in 2 independent cohorts: one consisted of 57 stage I-III CRC patients and 74 healthy individuals and another one with 47 stage IV patients and the same 74 healthy individuals. The model yielded a sensitivity of 83% and 95% for the early and late stage cohorts, respectively. A specificity of 95% was obtained for both cohorts. Conclusions: Our findings demonstrated the potential of profiling DNA methylation, which can effectively distinguish cancerous from healthy, for the purpose of screening. This method has potential to serve as a supplementary or alternative approach in early detection.

Early detection of colorectal cancer: A proteomic screening study for plasma biomarkers.

2012 ◽  
Vol 30 (4_suppl) ◽  
pp. 415-415
Author(s):  
Stine Buch Thorsen ◽  
Martin Lundberg ◽  
Erika Assarsson ◽  
Nick Gee ◽  
Mick Knowles ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Early Detection ◽  
Cross Reactivity ◽  
Plasma Samples ◽  
Regional Lymph Nodes ◽  
Cancer Disease ◽  
Dna Oligonucleotides ◽  
Early Markers ◽  
Screening Study ◽  
Specific Proteins

415 Background: Colorectal cancer (CRC) is the third most frequent cancer disease in the US. As the 5 year survival is closely associated to stage of disease at the time of diagnosis, patients with localized tumor limited to the colon or rectum have more than 90% chance of 5 year survival, while tumors spreading to regional lymph nodes or to distant organs, have a 5 year survival rate of 69% and 12%, respectively. Since late diagnosis is equal to a poor prognosis for the patients, biomarker guided early detection could make a difference for these patients. One strategy is blood based assays, where we hypothesis that a tumor and its microenvironment will lead to a deposition of specific proteins in the blood. However, due to the complexity of plasma, it is not only a biological but also a technical challenge to detect the proteins specific for CRC. Consequently, we combined the technical evaluation of multiplex proximity probing assays (PPA) with the search for relevant biomarkers for CRC. Each of the protein assays were based on thorough literature studies on the biology of CRC. Methods: Proteins were measured using multiplex PPA. Each assay was constructed from commercially available antibodies conjugated to two DNA oligonucleotides. When the antibodies from one assay bind the target protein simultaneously, it enables the DNA oligonucleotides to be either enzymatic ligated or extended to a PCR amplicon. This PCR amplicon reflects the identity of the proteins and can be quantified by real-time PCR. We evaluated the assays and searched for potential early markers using a collection of case-control plasma samples from a larger endoscopy study. In total we measure the levels of 150 different proteins in 296 human plasma samples from 74 CRC patients, 74 healthy individuals, 74 adenoma patients, and 74 patients with non-cancer diseases. Results: We have a successrate of 80%, the sensitivity is for most assays below 5 pM, and we find no cross-reactivity in chicken plasma for any of the assays. We previous reported that CEA, TIMP-1, CA242, and IL8 were upregulated in CRC plasma. These proteins were also identified in our extended study and the data will be presented. Conclusions: We find potential early markers, which we will validate in new sample material.

Detector-C: A Blood-based IVD with High Sensitivity and Specificity for Early Detection and Screening of Colorectal Cancer

2010 ◽  
Vol 48 (08) ◽  
Author(s):  
A Rosenthal ◽  
H Köppen ◽  
R Musikowski ◽  
R Schwanitz ◽  
J Behrendt ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Early Detection ◽  
Sensitivity And Specificity ◽  
High Sensitivity

Biomarkers for Early Detection of Colitis-associated Colorectal Cancer - Current Concepts, Future Trends

2020 ◽  
Vol 22 (1) ◽  
pp. 137-145
Author(s):  
Tomasz Mackiewicz ◽  
Aleksander Sowa ◽  
Jakub Fichna
Keyword(s):  
Colorectal Cancer ◽  
Early Detection ◽  
Cancer Development ◽  
Sporadic Colorectal Cancer ◽  
Future Trends ◽  
Significant Progress ◽  
Current Standard ◽  
Risk Of Cancer ◽  
Histological Testing ◽  
Made In

: Colitis-associated colorectal cancer (CAC) remains a critical complication of ulcerative colitis (UC) with mortality of approximately 15%, which makes early CAC diagnosis crucial. The current standard of surveillance, with repetitive colonoscopies and histological testing of biopsied mucosa samples is burdensome and expensive, and therefore less invasive methods and reliable biomarkers are needed. Significant progress has been made thanks to continuous extensive research in this field, however no clinically relevant biomarker has been established so far. This review of the current literature presents the genetic and molecular differences between CAC and sporadic colorectal cancer and covers progress made in the early detection of CAC carcinogenesis. It focuses on biomarkers under development, which can be easily tested in samples of body fluids or breath and, once made clinically available, will help to differentiate between progressors (UC patients who will develop dysplasia) from non-progressors and enable early intervention to decrease the risk of cancer development.

scholarly journals Colorectal cancer screening using a stool DNA-based SDC2 methylation test: a multicenter, prospective trial

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Chang Woo Kim ◽  
Hyunjin Kim ◽  
Hyoung Rae Kim ◽  
Bong-Hyeon Kye ◽  
Hyung Jin Kim ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Colon Cancer ◽  
Early Detection ◽  
Prospective Trial ◽  
Screening Colonoscopy ◽  
Quantitative Detection ◽  
Screening Programs ◽  
Crc Screening ◽  
Stool Dna ◽  
Dna 2

Abstract Background Prevention and early detection of colorectal cancer (CRC) is a global priority, with many countries conducting population-based CRC screening programs. Although colonoscopy is the most accurate diagnostic method for early CRC detection, adherence remains low because of its invasiveness and the need for extensive bowel preparation. Non-invasive fecal occult blood tests or fecal immunochemical tests are available; however, their sensitivity is relatively low. Syndecan-2 (SDC2) is a stool-based DNA methylation marker used for early detection of CRC. Using the EarlyTect™-Colon Cancer test, the sensitivity and specificity of SDC2 methylation in stool DNA for detecting CRC were previously demonstrated to be greater than 90%. Therefore, a larger trial to validate its use for CRC screening in asymptomatic populations is now required. Methods All participants will collect their stool (at least 20 g) before undergoing screening colonoscopy. The samples will be sent to a central laboratory for analysis. Stool DNA will be isolated using a GT Stool DNA Extraction kit, according to the manufacturer’s protocol. Before performing the methylation test, stool DNA (2 µg per reaction) will be treated with bisulfite, according to manufacturer’s instructions. SDC2 and COL2A1 control reactions will be performed in a single tube. The SDC2 methylation test will be performed using an AB 7500 Fast Real-time PCR system. CT values will be calculated using the 7500 software accompanying the instrument. Results from the EarlyTect™-Colon Cancer test will be compared against those obtained from colonoscopy and any corresponding diagnostic histopathology from clinically significant biopsied or subsequently excised lesions. Based on these results, participants will be divided into three groups: CRC, polyp, and negative. The following clinical data will be recorded for the participants: sex, age, colonoscopy results, and clinical stage (for CRC cases). Discussion This trial investigates the clinical performance of a device that allows quantitative detection of a single DNA marker, SDC2 methylation, in human stool DNA in asymptomatic populations. The results of this trial are expected to be beneficial for CRC screening and may help make colonoscopy a selective procedure used only in populations with a high risk of CRC. Trial registration: This trial (NCT04304131) was registered at ClinicalTrials.gov on March 11, 2020 and is available at https://clinicaltrials.gov/ct2/show/NCT04304131?cond=NCT04304131&draw=2&rank=1.

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