Colorectal Cancer Diagnostics Research Articles

Chemoprophylaxis of precancerous lesions in patients who are at a high risk of developing colorectal cancer (Review).

The diagnostics of colorectal cancer (CRC) and precancerous lesions in the colon is one of the most urgent matters to be considered for the modern protocols of complex examination, recommended for use from the age of 45 years, and including both instrumental and laboratory methods of research: Colonoscopy, CT colonography, flexible sigmoidoscopy, fecal occult blood test, fecal immunohistochemistry test and stool DNA test Nevertheless, the removal of those precancerous lesions does not solve the issue, and, apart from the regular endoscopic monitoring of patients who are at a high risk of developing CRC, the pharmacological treatment of certain key pathogenic mechanisms leading to the development of CRC is required. The present review to discusses the function of β-catenin in the transformation of precancerous colorectal lesions into CRC, when collaborating with PI3K/AKT/mTOR signaling pathway and other mechanisms. The existing methods for the early diagnostics and prevention of discovered anomalies are described and categorized. The analysis of the approaches to chemoprophylaxis of CRC, depending on the results of endoscopic, morphological and molecular-genetic tests, is presented.

Implications of single-cell immune landscape of tumor microenvironment for the colorectal cancer diagnostics and therapy.

Colorectal cancer (CRC) originates from the polyps lining the colon and is among the most common types of cancer. With the increasing popularity of single-cell sequencing technologies, researchers have been able to better understand the immune landscape of colorectal cancer, by analyzing their expression and interactions in detail with the tumor microenvironment (TME) at single-cell level. Since the tumor-immune cell interactions play a critical part in the advancement as well as treatment response in colorectal cancer, the release of inhibitory factors such as T cells are important for recognizing and destroying cancer cells. Such information is vital to identify immunotherapeutic targets for cure and monitoring response to treatments. Therefore, a comprehensive single-cell studies-based overview of key immunogenic agents regulating the TME of CRC is provided in this review. Tumor-associated macrophages can promote tumor growth and resistance to treatment by releasing factors that inhibit the function of other immune cells. Additionally, colorectal cancer cells can express programmed cell death protein 1 and its ligand, which can also inhibit T-cell function. Researchers have found that certain types of immune cells, prominently T cells, natural killer, and dendritic cells, can have a positive impact on the prognosis of colorectal cancer patients. Treatments like immune checkpoint inhibitors and CAR-T therapies that help to release the inhibitory signals from the cancer cells allow the immune cells to function more effectively.

Current Status and Emerging Trends in Colorectal Cancer Screening and Diagnostics.

Colorectal cancer (CRC) is a prevalent and potentially fatal disease categorized based on its high incidences and mortality rates, which raised the need for effective diagnostic strategies for the early detection and management of CRC. While there are several conventional cancer diagnostics available, they have certain limitations that hinder their effectiveness. Significant research efforts are currently being dedicated to elucidating novel methodologies that aim at comprehending the intricate molecular mechanism that underlies CRC. Recently, microfluidic diagnostics have emerged as a pivotal solution, offering non-invasive approaches to real-time monitoring of disease progression and treatment response. Microfluidic devices enable the integration of multiple sample preparation steps into a single platform, which speeds up processing and improves sensitivity. Such advancements in diagnostic technologies hold immense promise for revolutionizing the field of CRC diagnosis and enabling efficient detection and monitoring strategies. This article elucidates several of the latest developments in microfluidic technology for CRC diagnostics. In addition to the advancements in microfluidic technology for CRC diagnostics, the integration of artificial intelligence (AI) holds great promise for further enhancing diagnostic capabilities. Advancements in microfluidic systems and AI-driven approaches can revolutionize colorectal cancer diagnostics, offering accurate, efficient, and personalized strategies to improve patient outcomes and transform cancer management.

Mutational and DNA fragment analysis may help in the triage of FIT+ patients enrolled in colorectal cancer screening programs.

e15546 * F. Pepe made an equal contribution Background: Mutational and DNA fragment analysis have already been investigated for a possible role in the detection of colorectal cancer. This pilot study, conducted at ASL NA 3 SUD (Naples, Italy) with the University of Naples Federico II, attempts to understand their possible role in the detection of Advanced Adenomas+ (AA+) and their potential ability to triage subjects with a positive FIT test for referral to colonoscopy. Methods: 160 FIT+ subjects of both genders, aged 50-74, participating in a colorectal cancer screening program were enrolled in a clinical trial focusing on a novel biomarker for colorectal cancer diagnostics. Of these, 14 subjects had disease (12 AAs and 2 colorectal cancers, from now on “CRC”). After ethical committee’s approval and release of informed consents, a 20 ml whole-blood sample was collected from each subject. Samples were centrifuged following standard procedures and stored at -80°C. DNA extraction was later performed using a Minielute kit (Qiagen). Mutational and DNA fragment analysis were performed using a qPCR-based technology: the ColoScape™ 2.5 kit (DiaCarta Inc, Pleasanton, CA). A logistic regression was run to determine the probability of disease in subjects showing positivity for predictors. Estimates of sensitivity, specificity, PPV, NPV (with 95% C.I.), PLR, NLR and Youden’s Index were also obtained. Results: The logistic regression showed statistical significance, with a p-value < 0.0001, that is, the logistic regression model with provides a better fit than the model without the independent variable. ColoScape identified 12 subjects with disease out of 14, for a sensitivity of 86% (67%,100%); both CRCs were picked up by ColoScape together with 10 AAs. 2 AAs were missed. Out of 146 healthy subjects, 107 were successfully identified as negatives, for a specificity of 73% (66%, 80%). PPV was 24% (12%, 35%). NPV was 98% (96%, 100%). PLR, NLR and Youden’s Index were 3.21, 0.19 and 0.59 respectively. Conclusions: This pilot study was intended to provide performance indicators to design a larger population study that is currently underway. These preliminary findings suggest that liquid biopsy-based mutational and DNA fragment analysis can play a role in selecting subjects with a higher risk of returning a positive colonoscopy, thus potentially improving the quality of colorectal cancer screening programs and patients’ experience.

Unsupervised Learning Composite Network to Reduce Training Cost of Deep Learning Model for Colorectal Cancer Diagnosis

Deep learning facilitates complex medical data analysis and is increasingly being explored in colorectal cancer diagnostics. However, the training cost of the deep learning model limits its real-world medical utility. In this study, we present a composite network that combines deep learning and unsupervised K-means clustering algorithm (RK-net) for automatic processing of medical images. RK-net was more efficient in image refinement compared with manual screening and annotation. The training of a deep learning model for colorectal cancer diagnosis was accelerated by two times with utilization of RK-net-processed images. Better performance was observed in training loss and accuracy achievement as well. RK-net could be useful to refine medical images of the ever-expanding quantity and assist in subsequent construction of the artificial intelligence model.

Luminal and Tumor-Associated Gut Microbiome Features Linked to Precancerous Lesions Malignancy Risk: A Compositional Approach.

Colorectal cancer is the third most commonly diagnosed cancer worldwide. Human gut microbiome plays important roles in protecting against it, as well as contributing to its onset and progression. Identification of specific bacterial taxa associated with early stages of colorectal cancer may help develop effective microbiome-based diagnostics. For precancerous lesions, links of their characteristics to luminal and tumor-associated microbiome composition are to be elucidated. Paired stool and tumor brush biopsy samples were collected from 50 patients with precancerous lesions and early forms of colon cancer; their microbial communities were profiled using high-throughput 16S rRNA sequencing. We showed that the microbiome differences between stool and biopsy samples can be to a high extent computationally corrected. Compositionality-aware statistical analysis of microbiome composition revealed its associations with the number of lesions, lesion type, location and malignization pathway. A major determinant of precancerous lesions malignancy risk-the number of lesions-was positively associated with the abundance of H2S-producing taxa. Our results contribute to the basis for developing early non-invasive colorectal cancer diagnostics via identifying microorganisms likely participating in early stages of cancer pathogenesis.

Assessment of the Human Metabolome as a Method for Molecular Diagnostics of Colorectal Cancer: Prevention and Therapy

Assessment of the Human Metabolome as a Method for Molecular Diagnostics of Colorectal Cancer: Prevention and Therapy

Evaluation of combined detection of multigene mutation and SDC2/SFRP2 methylation in stool specimens for colorectal cancer early diagnosis

Molecular diagnostics of colorectal cancer (CRC) can be used as an auxiliary approach for patients recommended for colonoscopy, providing more CRC supplemental diagnosis options. This study investigated whether combined detection of KRAS/BRAF/APC mutation and SDC2/SFRP2 methylation can serve as auxiliary diagnostics in clinical management. KRAS/BRAF/APC mutation and SDC2/SFRP2 methylation in stool samples from healthy donors, patients with CRC, advanced adenoma (AA), non-advanced adenoma (NAA), or other gastroenterological diseases were evaluated using quantitative PCR (qPCR) or methylation-specific quantitative PCR (MSP). Test accuracy was determined by evaluating the tests' sensitivity, specificity, positive/negative predictive value (PPV/NPV), or positive/negative likelihood ratio (PLR/NLR). The combined fecal KRAS/BRAF/APC mutation and SFRP2/SDC2 methylation detection test achieved a sensitivity of 88.57% with a PPV of 93.64% and a PLR of 7.10 for CRC patients. In comparison, the corresponding parameters for multigene mutation were 46.67%, 92.59%, and 36.26 and 83.81%, 93.94%, and 7.47, for DNA methylation, separately. The sensitivity of the combined test, gene mutation test, and DNA methylation test approach was 75%, 28.26%, and 72.83%. Furthermore, the specificity of this approach in the NAA group was 79.49%. Meanwhile, the overall diagnostic specificity for the combined test in NAA, healthy control, and interference groups was 88.42%. In addition, the sensitivity of the combined detection method increased with the disease stage in CRC patients and elevated along with the lesion size (≥ 1cm) in AA patients. Combined detection of fecal KRAS/BRAF/APC mutation and SFRP2/SDC2 methylation has potential application value for the auxiliary diagnosis of CRC and AA.

The structure of synchronous multiple primary colorectal cancer

One of the problems of modern oncology is the lack of timely diagnosis of synchronous forms of colorectal cancer. Therefore, the aim of this study was to evaluate the structure of synchronous primary multiple forms of colorectal cancer. Methods: We analyzed the results of colorectal cancer diagnostics (n = 583) in the period 2010–2019. We did not include patients with metachronous colorectal cancer in the study. The average age of all patients was 71.6 ± 7.4 years. There were 1.5 times more men than women. Results: Synchronous colorectal cancer was detected in 171 (29.3%) patients. With synchronous primary multiple forms of colorectal cancer, 2 tumors were diagnosed in every second patient. In synchronous colorectal cancer, tumors were localized twice as often in the colon (n = 118) than in the rectum (n = 53). Conclusions: In patients with colorectal cancer, synchronous tumors occur in every third case. Synchronous colorectal cancer is more common in men. In the majority of patients (59.1%), synchronous colorectal cancer was detected within 6 months after imaging of the primary tumor.

Importance of Extracellular Vesicle Derived RNAs as Critical Colorectal Cancer Biomarkers.

Colorectal cancer typically begins from a nonmalignant polyp formation in the large intestine that, over time, develops into colorectal cancer. The growth of benign polyps can be checked if detected in the early stages of the disease. Doctors usually recommend colonoscopy to average and high-risk individuals for colorectal cancer screening. Elevated carcinoembryonic antigen (CEA) is a broadly used biomarker for colorectal cancer. The genetic and epigenetic alteration of genes such as p53, BRAF, APC, and PIK3CA is also correlated with colorectal cancer in various clinical studies. In general, tissue biopsy is most frequently used for colorectal cancer diagnosis, but the whole tumor heterogeneity cannot be accessed by this technique. Furthermore, such a highly invasive technique is not suitable for repeated testing. Recently, extracellular vesicles (EVs), lipid bilayer enclosed sacs secreted from colorectal cancer cells, are emerging as a diagnostic tool for colon cancer detection. The major advantages of using EVs for colon cancer diagnosis are (i) EVs can be isolated in a noninvasive manner from the body fluid and (ii) EV incorporated cargoes (mostly RNAs) reveal various aspects of colorectal cancer. EV-RNAs are also implicated in tumor invasion and influence the immune system for the further spread of tumors. However, due to the lack of standardized EV detection strategies, diagnostic applicability is limited. Herein, we review the recent literature on the pathobiological dependence of colorectal cancer on EV-RNAs. Further, we present the advantages of identification and characterization of EV-RNAs to explore the connection between differential expression of extracellular vesicle incorporated RNAs and colorectal cancer. How this approach may potentially translate into point of care colorectal cancer diagnostics is also discussed.

Transabdominal ultrasound diagnostics of intestinal tumor diseases in adults

Oncological diseases account for the largest share of all diseases of the colon, making up to 45% of all intestinal diseases. The colorectal cancer incidence is increasing every year in almost all countries, including Ukraine. Usually the endoscopic methods of examination and X-ray diagnostics play a key role in colon pathology diagnostics. Transabdominal ultrasound examination was considered a poor method to diagnose a cancer of hollow organs due to the presence of gas in the intestine. The potential possibilities of this method in assessment of the intestinal diseases as well as in studying the prevalence of pathological changes beyond the intestinal wall were underestimated. However, the development of ultrasound devices with high resolution determines the increasing use of transabdominal ultrasound examination to diagnose diseases of the gastrointestinal tract. The article is based on literature review as well as on authors’ own observations of ultrasound examination of benign and malignant diseases of the large intestine. The article goes on to analyze possibilities of transabdominal ultrasound diagnostics of pathological changes in the intestine. The results show that widespread use of ultrasound equipment, increasing the resolution of sensors can significantly complement the existing methods of colorectal cancer diagnostics. The use of transabdominal ultrasound examination in cases of colorectal cancer will allow to detect a colon cancer in the preclinical and mildly symptomatic stage, while excluding the influence of ionizing radiation.

Ex Vivo Vibration Spectroscopic Analysis of Colorectal Polyps for the Early Diagnosis of Colorectal Carcinoma.

Colorectal cancer is one of the most common and often fatal cancers in humans, but it has the highest chance of a cure if detected at an early precancerous stage. Carcinogenesis in the colon begins as an uncontrolled growth forming polyps. Some of these polyps can finally be converted to colon cancer. Early diagnosis of adenomatous polyps is the main approach for screening and preventing colorectal cancer, and vibration spectroscopy can be used for this purpose. This work is focused on evaluating FTIR and Raman spectroscopy as a tool in the ex vivo analysis of colorectal polyps, which could be important for the early diagnosis of colorectal carcinoma. Multivariate analyses (PCA and LDA) were used to assist the spectroscopic discrimination of normal colon tissue, as well as benign and malignant colon polyps. The spectra demonstrated evident differences in the characteristic bands of the main tissue constituents, i.e., proteins, nucleic acids, lipids, polysaccharides, etc. Suitable models for discriminating the three mentioned diagnostic groups were proposed based on multivariate analyses of the spectroscopic data. LDA classification was especially successful in the case of a combined set of 55 variables from the FTIR, FT Raman and dispersion Raman spectra. This model can be proposed for ex vivo colorectal cancer diagnostics in combination with the colonoscopic extraction of colon polyps for further testing. This pilot study is a precursor for the further evaluation of the diagnostic potential for the simultaneous in vivo application of colonoscopic Raman probes.

The prospective on molecular diagnostics of colorectal cancer and its clinical implications

The prospective on molecular diagnostics of colorectal cancer and its clinical implications

Colorectal malignant polyps: a modern approach.

Colorectal malignant polyps (MP) are polyps with invasive cancer into the submucosa harboring a variable risk of lymph node involvement, which can be estimated through evaluation of morphological, endoscopic, and histologic features. The recent advances in imaging endoscopic techniques have led to the possibility of performing an optical diagnosis of T1 colorectal cancer, allowing the selection of the best therapeutic modality to optimize outcomes for the patient. When MP are diagnosed after endoscopic removal, their management can be challenging. Differentiating low- and high-risk histologic features that influence the possibility of residual tumor, the risk of recurrence and the risk of lymph node metastasis, is crucial to further optimize treatment and surveillance plans. While the presence of high-risk features indicates a need for surgery in the majority of cases, location, comorbidities and the patient’s preference should be taken in account when making the final decision. This is a particularly important issue in the management of low rectal MP presenting with high-risk features, where chemoradiotherapy followed by a watch-and-wait strategy has demonstrated promising results. In this review we discuss the important prognostic features of MP and the most modern approaches regarding their management.

Diagnostic performance for T1 cancer in colorectal lesions ≥10mm by optical characterization using magnifying narrow-band imaging combined with magnifying chromoendoscopy; implications for optimized stratification by Japan Narrow-band Imaging Expert Team classification.

Magnifying narrow-band imaging (M-NBI) and magnifying chromoendoscopy (M-CE) enable accurate diagnosis of T1 colorectal cancer, but the diagnostic yields from combined M-NBI and CE have not been fully analyzed. We aimed to evaluate the diagnostic yield of combining Japan NBI Expert Team (JNET) classification using M-NBI and M-CE. Superficial colorectal lesions ≥10mm removed at a Japanese tertiary cancer center between February 2016 and December 2018 were included. We analyzed the relationship between JNET classification, M-CE findings, and histological results based on prospectively collected endoscopic and pathologic data. A total of 1573 lesions, including 56 superficial submucosal invasive cancers, 160 deep submucosal invasive cancers, and 81 advanced cancers (≥T2) were analyzed. The probability of deeply invasive cancer (95% confidence interval) was 1.8% (1.1-2.8), 30.1% (25.4-35.1), and 96.6% (91.5-99.1) in JNET Types 2A, 2B, and 3, respectively. The probability of deeply invasive cancer in JNET Type 2B lesions with non-V, VL, and VH pit pattern was 4.3%, 16.6%, 76.0%, respectively (P<0.001). Our study showed the stratification by M-NBI using JNET classification and the effect of additional M-CE for JNET Type 2B lesions.

Statistics assisted analysis of Raman spectra and imaging of human colon cell lines – Label free, spectroscopic diagnostics of colorectal cancer

The aim of the study was to evaluate the biochemical characteristics of normal CCD18-Co, CRL-1831 and cancer CaCo-2 human colon cell lines. Statistic assisted analysis of label-free Raman spectra demonstrates that normal and cancer human colon cells can be distinguished with high confidence based on vibrational properties. Moreover, vibrational spectra can be used to differentiate normal cells of epithelial CRL-1831 and fibroblast CCD18-Co origin with high sensitivity and specificity. Principal Component Analysis and Partial Least Squares Discriminant Analysis yielded effective and comparable cell classification based on vibrational features. We have proved that Raman spectroscopy is a powerful tool for cancer diagnostics, and could play an important clinical role for cell phenotyping.

A novel xenonucleic acid mediated molecular clamping technology for early colorectal cancer diagnostics.

e16106 Background: Colorectal cancer (CRC) is one of the leading causes of cancer-related death. Early detection is critical to reduce CRC morbidity and mortality. In order to meet this need, we developed a molecular clamping assay called ColoScape for early colorectal cancer diagnostics. Methods: Nineteen mutations in four genes APC, KRAS, BRAF and CTNNB1 associated with early events in CRC pathogenesis are targeted in the ColoScape assay. Xenonucleic Acid (XNA) mediated qPCR clamping technology was applied to minimize the wild-type background amplification in order to improve assay sensitivity of CRC mutation detection. The assay analytical performance was verified and validated, cfDNA and FFPE CRC patient samples were assessed for clinical performance, and a ROC curve was applied to evaluate its performance. Results: The data showed that the assay analytical sensitivity is 0.5% Variant Allele Frequency, corresponding to ~7-8 copies of mutant DNA with 5ng total DNA input per test. This assay is highly reproducible with intra-assay CV &lt; 3% and inter-assay CV &lt; 5%. We have investigated 380 clinical samples including plasma cfDNA and FFPE samples from patients with different stages of CRC. The preliminary assay clinical specificity and sensitivity for cfDNA were 100% (95% CI, 80.3-97.5%) and 92.2% (95% CI, 94.7-100%) respectively with AUC being about 0.96; and 96% (95% CI, 77.6-99.7%) specificity and 92% (95% CI, 86.1-95.6%) sensitivity with AUC 0.94 for FFPE. Conclusions: XNA mediated molecular clamping assay is a rapid, precise, and sensitive assay for the detection of CRC mutations in cfDNA or FFPE samples.

Identification of MicroRNAs as Biomarkers in Colorectal Cancer Diagnostics: An In silico approach

Colorectal cancer (CRC) is the most common malignancy in the gastrointestinal tract, the third most commonly diagnosed cancer, and a leading cause of cancer‐related deaths worldwide. The involvement of microRNAs in cancers plays a significant role in their pathogenesis. Specific expressions of these non‐coding RNAs also serve as biomarkers for early CRC diagnosis but their laboratory/molecular identification is challenging and expensive. Current diagnostic methods are invasive, more expensive, and lack specificity and sensitivity. Therefore the need for a less invasive, more sensitive and specific biomarkers for early detection of CRC. The present study aimed to identify potential microRNAs and their target genes for CRC diagnosis using in silico approach. Sequence similarity search was employed to obtain the candidate microRNA from two different datasets and target prediction tools were employed to determine their target genes. The involvement of these microRNAs was carried out through the intersection analysis between the candidate microRNAs target gene list and experimentally validated CRC expressed genes. This was further strengthened by determining their enrichment in DAVID. Gene ontology, pathway analysis, and protein network interactions were further evaluated using UniProt, and KEGG, and STRING respectively and visualized by Cytoscape. The cBioPortal was used to prioritize the candidate microRNAs target genes. Prognostic and expression analysis were finally performed on both the candidate microRNAs and the prioritized targets using PROGmiRV2, and PrognoScan, and SurvExpress, and FIREBROWSE respectively. This study, therefore, identified five candidate microRNAs, two hub genes (CTNNB1 and EGFR) and seven statistically significant target genes (p&lt; 0.05) associated with CRC. The molecular validation study of these microRNAs and their targets are pivotal to ascertain the biological fitness of these finding.

Capabilities of perfusion computed tomography in colorectal cancer detection and differential diagnosis of pathological conditions of the colon. Early results

The article presents the results of preoperative CT diagnostics of colorectal cancer using perfusion technique. It was found that CT perfusion can improve the diagnostics of local prevalence of colon cancer compared to standard computed tomography, as well as to differentiate colorectal cancer and inflammatory bowel disease.

Chitinase 3-Like 1, Nestin, and Testin Proteins as Novel Biomarkers of Potential Clinical Use in Colorectal Cancer: A Review.

Colorectal cancer is the third most commonly diagnosed cancer in males and the second most common in females. Only 10-20% of patients are diagnosed at the early stage of disease. Recently, the role of novel biomarkers of the neoplastic process in the early detection of colorectal cancer has been widely discussed. In this review, we focused on the three novel biomarkers that are of potential clinical importance in diagnosing and monitoring colorectal cancer. Chitinase 3-like 1 protein, also known as YKL-40, and nestin and testin proteins are produced by colorectal cancer cells. YKL-40 protein is a marker of proliferation, differentiation, and tissue morphogenetic changes. The level of YKL-40 is elevated in about 20% of patients with colorectal cancer. An increased expression of nestin indicates immaturity. It is a marker of angiogenesis in neoplastic processes. Testin protein is a component of cell-cell connections and focal adhesions. The protein is produced in normal human tissues, but not in tumor tissues. Downregulation of testin increases cell motility, spread, and proliferation, and decreases apoptosis. The usefulness and role of these biomarkers, both alone and combined, in the diagnostics of colorectal cancer should be further explored as early cancer detection may substantially improve treatment outcome and patient survival.