Colorectal Carcinoma Patients Research Articles

Individualized prediction of conditional survival for colorectal signet-ring cell carcinoma patients.

Conditional survival (CS) considers the time already survived after surgery and may provide additional survival information. The authors sought to construct and validate novel conditional survival nomograms for the prediction of conditional overall survival (OS) and cancer-specific survival (CSS) of colorectal signet-ring cell carcinoma (SRCC) patients. Patients diagnosed with stage I-III SRCC between 2010 and 2019 were identified from the Surveillance, Epidemiology, and End Results database. The formula calculating CS was: CS(x|y) = S(x+y)/S(x), where S(x) represents the survival at x years. CS nomograms were then constructed to predict the 5-year conditional OS and CSS, followed by internal validation. A total of 944 colorectal SRCC patients were finally identified in this study. The 5-year OS and CSS improved gradually with additional survival time. Univariate and multivariate Cox regression analysis conducted in training set revealed that age, race, T stage, LNR, and perineural invasion were independent risk factors for both OS and CSS. Two nomograms with considerable predictive ability were successfully constructed [area under the curve (AUC) for OS: 0.788; AUC for CSS: 0.847] and validated (AUC for OS: 0.773; AUC for CSS: 0.799) for the prediction of 5-year OS and CSS, based on the duration of 1-4 years post-surgery survival. The probability of achieving 5-year OS and 5-year CSS in colorectal SRCC patients improved gradually with additional time. Conditional nomograms considering survival time will be more reliable and informative for risk stratification and postoperative follow-up.

Microenvironment, systemic inflammatory response and tumor markers considering consensus molecular subtypes of colorectal cancer.

Introduction: Colorectal carcinomas (CRC) are one of the most frequent malignancies worldwide. Based on gene expression profile analysis, CRCs can be classified into four distinct subtypes also known as the consensus molecular subtypes (CMS), which predict biological behaviour. Besides CMS, several other aspects of tumor microenvironment (TME) and systemic inflammatory response (SIR) influence the outcome of CRC patients. TME and inflammation have important role in the immune (CMS1) and mesenchymal (CMS4) subtypes, however, the relationship between these and systemic inflammation has not been assessed yet. Our objective was to evaluate the connection between CMS, TME and SIR, and to analyze the correlation between these markers and routinely used tumor markers, such as CEA (Carcinoembryonic Antigen) and CA19-9 (Carbohydrate Antigen 19-9). Methods: FFPE (Formalin Fixed Paraffin Embedded) samples of 185 CRC patients were collected. TME was described using tumor-stroma ratio (TSR), Klintrup-Makinen (KM) grade, and Glasgow Microenvironment Score (GMS). CMS classification was performed on tissue microarray using MLH1, PMS2, MSH2 and MSH6, and pan-cytokeratin, CDX2, FRMD6, HTR2B and ZEB1 immunohistochemical stains. Pre-operative tumor marker levels and inflammatory markers [C-reactive protein - CRP, albumin, absolute neutrophil count (ANC), absolute lymphocyte count (ALC), absolute platelet count (APC)] and patient history were retrieved using MedSolution database. Results: Amongst TME-markers, TSR correlated most consistently with adverse clinicopathological features (p < 0.001) and overall survival (p < 0.001). Elevated CRP and modified Glasgow Prognostic Score (mGPS) were associated with worse outcome and aggressive phenotype, similarly to tumor markers CEA and CA19-9. Stroma-Tumor Marker score (STM score), a new combined score of CA19-9 and TSR delivered the second best prognostication after mGPS. Furthermore, CMS4 showed association with TSR and several laboratory markers (albumin and platelet derived factors), but not with other SIR descriptors. CMS did not show any association with CEA and CA19-9 tumor markers. Conclusion: More routinely available TME, SIR and tumor markers alone and in combination deliver reliable prognostic data for choosing the patients with higher risk for propagation. CMS4 is linked with high TSR and poor prognosis, but in overall, CMS-classification showed only limited effect on SIR- and tumor-markers.

Abstract LB252: iCellSight: An interactive tool for analyzing and visualizing in-situ high-plex cellular protein data

Abstract Introduction The increased use of tissue-based multiplex immunohistochemistry (mIHC) in cancer research has significantly enhanced the generation of spatial hyper-plex images, crucial for understanding complex tumor-immune microenvironments. This advancement has led to a growing demand for spatial data analysis tools that enable quick and insightful evaluation. While existing spatial analysis software like Steinbock and SPIAT offer foundational support, effectively managing the high-dimensional and voluminous cellular data to derive meaningful biological insights remains challenging. This is especially true for researchers lacking bioinformatics expertise who seek to conduct quick preliminary evaluations. To address this gap, we introduce iCellSight: a user-friendly, code-free interactive analysis and visualization tool tailored to empower researchers with rapid and intuitive interpretation of in-situ cellular data. Material and methods A formalin-fixed, paraffin-embedded colorectal carcinoma (CRC) patient tissue section was subjected to staining using COMET (Lunaphore, Switzerland), which automates sequential immunofluorescence staining and imaging of up to 40 protein markers. The stacked image file produced was analyzed with HALO (Indica Labs, USA), which generated cell-level data like cell coordinates, stain intensity values and user-defined cell types. We developed a Shiny App tool, using the Shiny R package, with key features for cell data interpretation: (1) user-input cell number subsampling; (2) dimensional reduction plots with PhenoGraph for unsupervised clustering, allowing a choice of UMAP or t-SNE, and neighbor number selection of 30, 50 or 150; (3) 2D tissue space mapping of PhenoGraph clusters; and (4) heatmap visualizations displaying cluster-wise biomarker expression (with customizable color schemes and scaling), biomarker co-expression, and cluster pair-wise distance. Results Using iCellSight, 297,000 cells were detected from the CRC sample, where 27 clusters were identified. We observed 3 distinct clusters from the t-SNE plot that consist of macrophages, granulocytes, and NK cells and monocytes respectively. From the cluster projection in 2D tissue space, the cluster of NK cells and monocytes was found to be densely populated in a particular area of the tumor nest. Conclusion We presented iCellSight, a code-free visualization and analysis tool which streamlines the semi-automated analysis of spatial hyper-plex protein data. Designed with an intuitive user interface, this app enables researchers to efficiently navigate its features without requiring any prior coding expertise. We anticipate that the visualizations and analysis provided by this tool will significantly assist researchers in their data analyses and facilitate the validation of experimental hypotheses. Citation Format: Yancun Zhu, Felicia Wee, Menaka Priyadharsani Rajapakse, Esther Gek Teo, Nicholas Ang, Solomonraj Wilson, Zheng Yi Ho, Willa Wen-You Yim, Li Yen Chong, Craig Ryan Joseph, Jeffrey Chun Lim, Zhen Wei Neo, Chwee Ming Lim, Bernett Lee, Olaf Rotzschke, Joe Yeong, Mai Chan Lau. iCellSight: An interactive tool for analyzing and visualizing in-situ high-plex cellular protein data [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 2 (Late-Breaking, Clinical Trial, and Invited Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(7_Suppl):Abstract nr LB252.

Lectin-Based Immunophenotyping and Whole Proteomic Profiling of CT-26 Colon Carcinoma Murine Model.

A murine colorectal carcinoma (CRC) model was established. CT26 colon carcinoma cells were injected into BALB/c mice's spleen to study the primary tumor and the mechanisms of cell spread of colon cancer to the liver. The CRC was verified by the immunohistochemistry of Pan Cytokeratin and Vimentin expression. Immunophenotyping of leukocytes isolated from CRC-bearing BALB/c mice or healthy controls, such as CD19+ B cells, CD11+ myeloid cells, and CD3+ T cells, was carried out using fluorochrome-labeled lectins. The binding of six lectins to white blood cells, such as galectin-1 (Gal1), siglec-1 (Sig1), Sambucus nigra lectin (SNA), Aleuria aurantia lectin (AAL), Phytolacca americana lectin (PWM), and galectin-3 (Gal3), was assayed. Flow cytometric analysis of the splenocytes revealed the increased binding of SNA, and AAL to CD3 + T cells and CD11b myeloid cells; and increased siglec-1 and AAL binding to CD19 B cells of the tumor-bearing mice. The whole proteomic analysis of the established CRC-bearing liver and spleen versus healthy tissues identified differentially expressed proteins, characteristic of the primary or secondary CRC tissues. KEGG Gene Ontology bioinformatic analysis delineated the established murine CRC characteristic protein interaction networks, biological pathways, and cellular processes involved in CRC. Galectin-1 and S100A4 were identified as upregulated proteins in the primary and secondary CT26 tumor tissues, and these were previously reported to contribute to the poor prognosis of CRC patients. Modelling the development of liver colonization of CRC by the injection of CT26 cells into the spleen may facilitate the understanding of carcinogenesis in human CRC and contribute to the development of novel therapeutic strategies.

Insights Into Colorectal Carcinoma: A Comprehensive Review of MicroRNA Expression Patterns.

Colorectal carcinoma (CRC) remains a significant contributor to cancer-related morbidity and mortality worldwide. MicroRNAs (miRNAs) have emerged as crucial regulators of gene expression and play critical roles in various biological processes, including carcinogenesis. This comprehensive review aims to elucidate the role of miRNAs in CRC by analyzing their expression patterns and functional implications. An extensive literature review identified dysregulated miRNAs associated with differentstages of CRC progression, from initiation to metastasis. These miRNAs modulate key signaling pathways in tumor growth, invasion, and metastasis. Furthermore, we discuss the potential of miRNAs as diagnostic biomarkers and therapeutic targets in CRC management. Future research directions include elucidating the functional significance of dysregulated miRNAs using advanced experimental models and computational approaches and exploring the therapeutic potential of miRNA-based interventions in personalized treatment strategies for CRC patients. Collaboration among researchers, clinicians, and industry partners will be essential to translate these findings into clinically impactful interventions that improve patient outcomes in CRC.

Abstract 4065: Entinostat synergizes with the IL-15 superagonist N-803 and anti-PD-1 to engage cDC1 and stem-like TCF-1+ CD8+ T and NK cells to overcome colorectal tumors refractory to anti-PD-1 therapy

Abstract Background: Immune checkpoint blockade (ICB) has limited efficacy in colorectal carcinoma (CRC), where ~85% of patients harbor microsatellite stability (MSS). A minor subset of CRC patients (~15%) harbor microsatellite instability (MSI) and demonstrate improved responses to ICB ranging from 30-50%. Combination therapy has the potential to increase the response rate. Entinostat, a class I HDAC inhibitor, has demonstrated potential to enhance ICB efficacy, such as αPD-1 and immunostimulatory cytokines. N-803, a heterodimeric IL-15 superagonist that enhances proliferation and activation of CD8+ and NK cells has demonstrated to enhance the clinical benefit of Nivolumab in non-small cell lung cancer patients that were relapsed or refractory to αPD-1 monotherapy. Here, we investigated the antitumor efficacy and mode of action of combination therapy comprising αPD1, Entinostat, and N-803 in αPD1-refractory CT26 and Jak1-deleted MC38 CRC tumor models. Methods: Entinostat, N-803, and αPD1 alone or in combination were administered to mice bearing established CT26 or Jak1-deleted MC38 tumors. Antitumor efficacy, survival, and protective memory upon tumor rechallenge were monitored. Comprehensive flow cytometric profiling of the tumor microenvironment (TME), tumor-draining lymph node (tdLN), and spleen cell populations was conducted. CT26 tumor and tdLN immunomes were examined by single-cell transcriptomics. Cytokines and chemokines were examined in the TME and sera. Antigen-specific IFNg T cell responses were evaluated by ELISpot. Results: We demonstrated that combination therapy with Entinostat, N-803 and αPD1 elicits potent suppression of CT26 and Jak1-deleted MC38 tumors, with median overall survival increasing 33% to not reached. These effects were associated with increased activation, infiltration, cytokine production, and cytolytic function of tumor CD8+ T and NK cells, and increased sera IFNg and TNFα. Tumor resolution and protective memory were associated with development of p15e-specific T cell responses. Increased migration of CD8+ T cells and cross-presenting dendritic cells (cDC1) to the tdLN, as well as significant enrichment of cross-presentation genes in tumor cDC1s was observed. Transcriptomic and phenotypic analyses demonstrated increased stem-like TCF-1+ CD8+ T cell accumulation in the tdLNs and TME of mice treated with triple combination. Conclusions: Combination therapy with Entinostat plus N-803 and αPD1 synergized to promote a sustained antitumor response and prolonged survival. We hypothesize the antitumor effects were due to collaborative efforts of CD8+ T and NK cells, and cDC1 cells in tdLN and TME. These results provide a rationale for the clinical testing of αPD1, Entinostat and N-803 combination for patients with CRC, including those with acquired resistance to ICB. Citation Format: Christine M. Minnar, Masaya Miyamoto, Asma S. Khelifa, Grace Lui, Katherine L. Lothstein, Paul L. Chariou, Thomas J. Meyer, Alexei Lobanov, Margaret Cam, Jeffrey Schlom, Sofia R. Gameiro. Entinostat synergizes with the IL-15 superagonist N-803 and anti-PD-1 to engage cDC1 and stem-like TCF-1+ CD8+ T and NK cells to overcome colorectal tumors refractory to anti-PD-1 therapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 4065.

Abstract 2814: Rectal implantation paves the path to evaluate the metabolic machinery responsible for CRC metastasis

Abstract Introduction: Colorectal carcinoma patients die most often from distant metastases in liver and lung. It is crucial to develop metastatic models for the CRC in order to better understand the process, which will help in treatment. Method: Using orthotropic transplantation techniques, we tried to test whether rectal implantation could produce spontaneous metastases in-vivo mimicking colorectal cancer. The rectum was injected orthotopically with 0.2 × 106 HCT116 cells. In order to confirm metastasis, histopathological examinations were performed. Further, we generated cancer organoids to evaluate population susceptibility to CRC metastasis and metabolic mechanisms responsible for metastasis process. Results: We found that rectal implantation was an effective method for simulating CRC cancer metastasis using H&amp;E staining and immunohistochemistry. In less than four weeks after the implant, HCT116 spontaneously metastasized to the liver, lungs, and D lymph node. When compared with cancer organoids derived from primary tumors, liver and lung metastases organoids proliferated more often, were more aggressive, and displayed distinct characteristics. Using non-targeting metabolomic analysis, it was found that metastases derived organoids and primary tumors derived organoids were distinguished by novel metabolites. Conclusion: By implantation in the rectal cavity, we can evaluate the metabolic machinery that leads to CRC metastasis. In the future, implantation of patient-derived samples will become increasingly important. Citation Format: Said M. Afify, Chen Shen, Siva B. Merugu, Daniel T. Wynn, Anmada Nayak, Wenhui Lu, Ricardo M. Calderin, Evan Pannkuk, Sung H. Hong, Nagi G. Ayad, David J. Robbins. Rectal implantation paves the path to evaluate the metabolic machinery responsible for CRC metastasis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 2814.

Alleviating misclassified germline variants in underrepresented populations: A strategy using popmax

PurposeGermline variant interpretation often depends on population-matched control cohorts. This is not feasible for population groups that are underrepresented in current population reference databases. MethodsWe classify germline variants with population-matched controls for 2 ancestrally diverse cohorts of patients: 132 early-onset or familial colorectal carcinoma patients from Singapore and 100 early-onset colorectal carcinoma patients from the United States. The effects of using a population-mismatched control cohort are simulated by swapping the control cohorts used for each patient cohort, with or without the popmax computational strategy. ResultsPopulation-matched classifications revealed a combined 62 pathogenic or likely pathogenic (P/LP) variants in 34 genes across both cohorts. Using a population-mismatched control cohort resulted in misclassification of non-P/LP variants as P/LP, driven by the absence of ancestry-specific rare variants in the control cohort. Popmax was more effective in alleviating misclassifications for the Singapore cohort than the US cohort. ConclusionUnderrepresented population groups can suffer from higher rates of false-positive P/LP results. Popmax can partially alleviate these misclassifications, but its efficacy still depends on the degree with which the population groups are represented in the control cohort.

Association of Microsatellite Instability and Gene Expression Profile in Colorectal Carcinoma and Potential Implications for Therapy.

Background and Objective: In sporadic colorectal carcinomas (CRC), microsatellite instability (MSI) pathways play important roles. Previously, we showed differences in DNA methylation patterns in microsatellite stable (MSS) colorectal carcinomas and MSI-CRC. In the current study, we explore the similarities and differences in gene expression profiles in MSS and MSI at the gene level and at the pathway level to better understand CRC pathogenesis and/or the potential for therapeutic opportunities. Material and Methods: Seventy-one CRC patients (MSI = 18, MSS = 53) were studied. Paired tumor and adjacent normal tissues were used for genome-wide gene expression assays. Result: At the gene level, we compared the list of differentially expressed genes (fold change (FC) ≥ 3 and FDR < 0.05) in tumor tissues compared to corresponding normal tissue in CRC patients with MSI tumors (190 genes) and MSS tumors (129 genes). Of these, 107 genes overlapped. The list of genes that were differentially expressed in MSI tumors only showed enrichment predominantly in two broad categories of pathways-(a) Inflammation-related pathways including the interleukin-17 (IL-17) signaling pathway, tumor necrosis factor (TNF) signaling pathway, chemokine signaling, nuclear factor kappa B (NFκB) signaling, and cytokine-cytokine interactions, and (b) metabolism-related pathways, including retinol metabolism, steroid hormone biosynthesis, drug metabolism, pentose and glucoronate interconversions, and ascorbate and aldarate metabolism. The genes in inflammation-related pathways were up-regulated whereas genes in metabolism-related pathways were down-regulated in MSI tumor tissue. Pathway-level analysis also revealed similar results confirming the gene enrichment findings. For example, the 150 genes involved in the IL-17 signaling pathway were on average up-regulated by 1.19 fold (CI 1.16-1.21) in MSI compared to 1.14 fold (CI 1.13-1.16) in MSS patients (interaction p = 0.0009). Conclusions: We document an association between MSI status and differential gene expression that broadens our understanding of CRC pathogenesis. Furthermore, targeting one or more of these dysregulated pathways could provide the basis for improved therapies for MSI and MSS CRC.

Clinicopathological and Immunohistochemical Association of CD55 and CD59 in Colorectal Carcinoma: A Cross-sectional Study

Introduction: Colorectal Carcinoma (CRC) is one of the significant causes of fatality worldwide. High-grade and highstage cancers contribute to this fatality. In most terminal cases, new immunomarkers (CD55 and CD59) are commonly detected as positive. The expression of these immunomarkers and their clinical relevance in CRC has not yet been firmly established. While their upregulation has been demonstrated in some CRC several groups have also reported that they are not highly expressed in CRC tissues. Aim: To study the expression of CD55 and CD59 in CRC. Materials and Methods: This was a cross-sectional study involving 90 patients with colorectal growth, conducted from February 2020 to July 2021 in the Department of Pathology in collaboration with the Department of General Surgery at Nil Ratan Sircar Medical College, Kolkata, West Bengal, India. Histopathological findings were evaluated, and associations of grading, staging, tumour type, tumour location, age, and sex were studied with immunomarkers (CD55 and CD59). Clinical parameters of the patients with CRC diagnosed by colonoscopic biopsy were also studied, including age, sex, clinical pictures, food habits, family history of CRC, and prior chemotherapy. The Chi-square test was used to determine the significance of the study. Data were analysed using the Statistical Package for the Social Sciences (SPSS) version 27.0 (IBM, Illinois, US). A p-value of ≤0.05 was considered statistically significant. Results: A total of 90 CRC patients were included in this study, with the majority being male 56 (62.2%) aged 51-60 years. Most cases 59 (65.6%) were moderately differentiated adenocarcinoma, followed by poorly differentiated adenocarcinoma 18 (20%) and well-differentiated adenocarcinoma 13 (14.4%). About 62 (68.9%) and 28 (31.1%) patients showed strong and weak CD55 expression, respectively. In total, 65 (72.2%) and 25 (27.8%) patients exhibited strong and weak CD59 expression, respectively. The association of CRC grade and stage with CD55 and CD59 was significant (p-value=0.0001, p-value=0.0013, p-value=0.0001, and p-value=0.0001, respectively). Conclusion: There are several variables to consider during the histopathological reporting of CRC, with tumour differentiation, grade, and stage being among the most important. Particularly, high-grade (poorly differentiated) and high-stage adenocarcinomas demonstrate enhanced expression of CD55 and CD59, indicating a poor prognosis. It is essential for pathologists to meticulously perform grossing and reporting of CRC and dispatch the histopathology report after proper clinicopathological correlations. These immunomarkers can also be included in a routine IHC panel for prognostic and therapeutic purposes.

Diagnostic Utility of Beta-Catenin in Poorly Differentiated Colorectal Carcinoma

Objective: To determine the diagnostic ability of Beta-catenin in poorly differentiated colorectal carcinomas. Study Design: Cross-sectional study Place and Duration of Study: Department of Histopathology PNS Shifa Hospital, Karachi Pakistan from Jun 2019 to Jun 2020 Methodology: After ethical approval from the Institutional Review Board, 60 patients of both gender and all age group, diagnosed with a case of colorectal carcinoma on biopsy and resection specimens analyzed on histopathology on H &amp; E staining at PNS SHIFA were included in the study. The resection specimens of CRC included those obtained from hemicolectomy, abdominoperineal resection and biopsies. Beta-catenin was interpreted using immunohistochemistry as cytoplasmic and nuclear staining with varying intensity. The scoring method was 0 as negative, 1+ as weakly positive, 2+ as moderate positive, and 3+ as strongly positive. Results: Among the 60 patients with resection specimens included from hemicolectomy, abdominoperineal resection and colorectal biopsies, strong nuclear positive results were observed in 30(50 %) patients, moderate nuclear positivity in 12 (20 %), weak nuclear positivity in 6(10 %) and positive cytoplasmic staining in 12(20 %) patients. Conclusion: Beta-catenin can be used successfully as a diagnostic utility in poorly differentiated colorectal carcinoma patients.    

Prognostic potential of MNX1-AS1 in chemotherapy of colorectal carcinoma.

To clarify the role of MNX1-AS1 in 5-FU resistance of Colorectal carcinoma (CRC). Relative levels of MNX1-AS1 in CRC and paracancerous tissues were detected by quantitative real-time polymerase chain reaction (qRT-PCR). Recruited CRC patients were treated by 5-FU-based FOLFOX chemotherapy, and they were divided to effective group and non-effective group according to the therapeutic efficacy, followed by comparison of their differences in clinical indicators. Influences of MNX1-AS1 on clinical features of CRC were analyzed. In addition, in vitro level of MNX1-AS1 in 5-FU-resistant HCT-8 cells and their parental cells was detected. After knockdown of MNX1-AS1 in HCT-8/5-FU cells, viability change was evaluated by cell counting kit-8 (CCK-8) assay. At last, regulatory effects of MNX1-AS1 on expression levels of ABC family genes were detected. MNX1-AS1 was upregulated in CRC tissues than paracancerous ones, and its level was higher in 5-FU-resistant CRC cases in comparison to 5-FU-sensitive cases. MNX1-AS1 level was linked to tumor size, tumor differentiation, depth of invasion, TNM staging and lymphatic metastasis in CRC. Notably, TNM staging, depth of invasion and lymphatic metastasis could affect the efficacy of FOLFOX chemotherapy in CRC patients. Knockdown of MNX1-AS1 reduced viability in HCT-8/5-FU cells, and downregulated ABCA1, ABCB1, ABCC1, ABCG1 and ABCG2. MNX1-AS1 triggers 5-FU resistance in CRC cells. Knockdown of MNXK1-AS1 is conductive to the well response to FOLFOX chemotherapy in CRC patients.

Evaluation of HHLA2 and CD8 Immunohistochemical Expression in Colorectal Carcinoma and Their Prognostic Significance.

Colorectal carcinoma (CRC) is the third most common malignancy worldwide. Human endogenous retrovirus H long terminal repeat-associating protein 2 (HHLA2) is a novel immune checkpoint molecule. The association between HHLA2 expression and clinicopathological features and its prognostic significance in CRC patients are still controversial. The aim of this study is to evaluate the prognostic value of immunohistochemical (IHC) expression of HHLA2 and CD8 in CRC. This retrospective study included 134 cases diagnosed with primary CRC at the Gastrointestinal Surgery Center (GISC) department, Mansoura Faculty of Medicine, during the period from December 2014 to December 2018. Clinicopathological and survival data were collected. IHC for HHLA2 and CD8 was performed, and they were correlated with clinicopathological parameters and patient prognosis. Among 134 CRC cases, high HHLA2 expression was detected in 73 (54.5%). High HHLA2 expression was significantly related to the depth of invasion (P = 0.005*), lymph node metastasis (P = 0.01*), tumor stage )P = 0.002*), and distant recurrence )P = 0.012*). Multivariate analysis spotted HHLA2 high expression as an independent prognostic predictor for OS in CRC (P = 0.03*) and DFS (P = 0.008*). CD8 shows a significant correlation with tumor infiltrating lymphocytes (TILs) (P ≤ 0.001*), absence of metastasis ((P = 0.029*), absence of tumor deposits (P=0.014*). However, CD8 shows no significant association with survival or HHLA2. HHLA2 is an independent prognostic factor for the overall survival and disease free survival of CRC patients and can predict poor prognosis in CRC patients.

A nomogram for the prediction of survival for colorectal signet ring cell carcinoma after surgery: A population-based study.

The aim was to construct and verify a nomogram-based assessment of cancer-specific survival (CSS) in patients with colorectal signet ring cell carcinoma after surgery. Patients were collected from Surveillance, Epidemiology, and End Results program between 2004 and 2015. Independent prognostic indicators were determined in the training cohort by Cox regression model. We identified 2217 eligible patients, who were further categorized into the training set (n = 1693) as well as the validation set (n = 524). Multivariate analysis revealed that age at diagnosis, gender, grade, tumor size, T stage, N stage, and M stage were independent predictive indicators. Then, the above 7 predictive factors were incorporated into a nomogram model to assess CSS, which showed good calibration and discrimination capacities in both sets. Both internal and external calibration plot diagrams revealed that the actual results were consistent with the predicted outcomes. The time-independent area under the curves for 3-year and 5-year CSS in the nomogram were larger than American Joint Committee on Cancer and Surveillance, Epidemiology, and End Results summary stage system. Moreover, decision curve analysis indicated the clinical utility of the nomogram. The nomogram demonstrated favorable predictive accuracy of survival in colorectal signet ring cell carcinoma patients after surgery, which should be further confirmed before clinical implementation.

[Tumor-immune cell interaction and senescence-associated molecules in colorectal carcinoma].

Cellular senescence permanently arrests the cell cycle of premalignant cells following protumorigenic stimuli, counteracting tumor progression. Senescence induction leads to phenotypic and metabolic changes and alters the interaction with the cells' microenvironment. This mediates tumor immunosurveillance but bears promalignant potential and may contribute to disease progression. Our study aims to investigate the prognostic potential of senescence markers in colorectal carcinoma (CRC) and to understand the interaction of senescent tumor cells and immune cells. Immunohistochemical markers were studied on atissue microarray (TMA) containing tumor tissue of n = 598 CRC patients and were evaluated using digital image analysis. Results were correlated with disease-specific survival (DSS) and progression-free survival (PFS). Consecutive TMA sections were stained for senescence markers and immune cell markers to analyze the spatial relation of those cell populations. Senescence was induced in CRC cell lines in vitro and co-cultures with various immune cell lines were established to study the interactions. Expression of different senescent-associated markers is associated with increased or decreased DSS and PFS. Close proximity of p21+ senescent tumor cells and CD8+ immune cells correlates with increased DSS and PFS. In vitro, senescent cells were dose-dependently eliminated by immune cells, which is facilitated via direct cell-cell contact and induction of apoptosis. Depicting the initiation of this important anti-tumor mechanism, markers of cellular senescence are of significant prognostic relevance in CRC. Moreover, our results show the pleiotropic effect of senescence in vivo. Absence as well as exceeding expression of senescence markers are associated with anegative prognosis in CRC. The impact of cellular senescence depends on the tumor microenvironment and the immunosurveillance of senescent cells. Proximity analyses of senescent cells and tumor-infiltrating immune cells have significant prognostic relevance and reflect this.

Is Metformin Associated with a Lower Prevalence of Polyps, Adenomas, and Colorectal Carcinoma in Patients with Diabetes Mellitus?

Recent studies suggested a protective role of metformin in the development of colorectal cancer (CRC) and its precursors. We aimed to investigate if metformin was associated with a lower prevalence and number of colorectal polyps in diabetic patients and also adenomas, high-risk adenomas, and CRC. Retrospective study on adult patients with diabetes mellitus followed in our hospital with a total colonoscopy between 2015 and 2019, treated with either metformin for > 5years or other antidiabetic agent (control group). We assessed the number, size, and histopathology examination of proliferative lesions detected on colonoscopy. We included 401 patients aged 69 ± 9years, 57% males, divided into two groups: treated with metformin (n = 260) and without (n = 141). The number of polyps detected was significantly lower in patients under metformin (p = 0.014). There was a nonsignificant trend towards lower polyp detection rates in the metformin compared to the control group both in unadjusted analysis (50% vs 60%, p = 0.058) and multivariable adjusted analysis (odds ratio [OR] 0.68, 95% confidence interval [CI] 0.43-1.09, p = 0.111). In the latter, we identified male gender (OR 2.24, 95%CI 1.44-3.49, p < 0.001), age (OR 1.35 for every 10years, 95%CI 1.07-1.71, p = 0.012), glycated hemoglobin value (OR 1.20 for every 1% increase, 95%CI 1.06-1.37, p = 0.005), and hypertension (OR 1.76, 95%CI 1.01-3.08, p = 0.046) as factors associated with a higher prevalence of polyps. We saw no statistically significant differences regarding adenoma (p = 0.231), high-risk adenoma (p = 0.810), and CRC (p = 0.705) diagnoses between groups. In our study, metformin was associated with less colorectal polyps in diabetic patients compared to other treatment modalities. We observed a nonsignificant trend towards lower polyp detection rates in the metformin group both in unadjusted and adjusted analyses.

Utility of CD34 in Assessing Microvessel Density and Its Correlation With Clinicopathological Parameters in Colorectal Carcinoma Patients.

Currently, the most commonly practiced method of reporting cases of colorectal carcinoma is done according to guidelines provided by the College of American Pathologists (8th edition) and the Royal College of Pathologists (UK). These guidelines include various histopathological parameters like tumor site, extent, histologic type, grade, margins, tumor budding, lymphovascular invasion, and perineural invasion. However, in the present guidelines, the immunohistochemistry-based marker of mean vessel density (MVD) has not been addressed as an important parameter. The present study gives an overview of the importance of MVD.MVD was statistically significant when correlated with tumor size, lymph node metastasis, grade, and vascular invasion. However, no statistical significance was observed when compared with age, perineural invasion, and stage of the tumor.

Lymph Node Molecular Analysis with OSNA Enables the Identification of pT1 CRC Patients at Risk of Recurrence: A Multicentre Study.

Early-stage colorectal carcinoma (CRC)-pT1-is a therapeutic challenge and presents some histological features related to lymph node metastasis (LNM). A significant proportion of pT1 CRCs are treated surgically, resulting in a non-negligible surgical-associated mortality rate of 1.5-2%. Among these cases, approximately 6-16% exhibit LNM, but the impact on survival is unclear. Therefore, there is an unmet need to establish an objective and reliable lymph node (LN) staging method to optimise the therapeutic management of pT1 CRC patients and to avoid overtreating or undertreating them. In this multicentre study, 89 patients with pT1 CRC were included. All histological features associated with LNM were evaluated. LNs were assessed using two methods, One-Step Nucleic Acid Amplification (OSNA) and the conventional FFPE plus haematoxylin and eosin (H&E) staining. OSNA is an RT-PCR-based method for amplifying CK19 mRNA. Our aim was to assess the performance of OSNA and H&E in evaluating LNs to identify patients at risk of recurrence and to optimise their clinical management. We observed an 80.9% concordance in LN assessment using the two methods. In 9% of cases, LNs were found to be positive using H&E, and in 24.7% of cases, LNs were found to be positive using OSNA. The OSNA results are provided as the total tumour load (TTL), defined as the total tumour burden present in all the LNs of a surgical specimen. In CRC, a TTL ≥ 6000 CK19 m-RNA copies/µL is associated with poor prognosis. Three patients had TTL > 6000 copies/μL, which was associated with higher tumour budding. The discrepancies observed between the OSNA and H&E results were mostly attributed to tumour allocation bias. We concluded that LN assessment with OSNA enables the identification of pT1 CRC patients at some risk of recurrence and helps to optimise their clinical management.

BRD4-binding enhancer promotes CRC progression by interacting with YY1 to activate the Wnt pathway through upregulation of TCF7L2

Colorectal carcinoma (CRC), one of the most life-threatening cancer types, is associated with aberrant expression of epigenetic modifiers and activation of the Wnt pathway. However, the role of epigenetic regulators in driving cancer cell proliferation and their potential as therapeutic targets affecting the Wnt pathway remain unclear. In this study, BRD4 was found to promote the progression of CRC both in vitro and in vivo. The expression of BRD4 correlated with shortened CRC patient survival. In addition, BRD4 function was strongly correlated with the Wnt pathway, but rather through regulation of TCF7L2 at transcriptional levels. BRD4 and H3K27ac have overlapping occupancies in the cis-regulatory elements of TCF7L2, suggesting enhancer-based epigenetic regulation. Numerous YY1 binding sites were found in the abovementioned region. YY1 recruited BRD4 to bind to cis-regulatory elements of TCF7L2, thereby regulating the expression of TCF7L2. Altogether, this study validates that BRD4 performs a canonical epigenetic regulatory function in CRC and can be used in the treatment of Wnt pathway-dependent CRC or other malignancies with clinically available bromodomain inhibitors.

Biliary Carcinoembryonic Antigen (CEA) Levels: The Role in Detection of Occult Hepatic Metastases in Colorectal Carcinoma

Objective: Colorectal carcinoma (CRC) is a major public health concern, often complicated by hepatic metastasis. Despite the widespread use of serum Carcinoembryonic Antigen (CEA) for postoperative monitoring, early detection of hepatic metastasis remains elusive. This study aims to evaluate the prognostic significance of CEA levels in both peripheral blood and gallbladder bile for predicting hepatic metastasis in CRC patients, and to explore its potential utility for personalized treatment regimens. Materials and Methods: A prospective randomized study was conducted over three years, enrolling 31 participants, 21 of whom underwent curative operations for CRC. Preoperative and postoperative CEA levels were assessed with rigorous diagnostic imaging and histological examinations. Patients were stratified into groups based on liver metastasis, postoperative chemotherapy, and CEA levels in both blood and bile. Results: A statistically significant difference was found in biliary CEA levels between patients with (mean = 129.22 ng/ml) and without intraoperative liver metastasis (mean = 33.16 ng/ml), p-value = 0.01. Serum and biliary CEA levels were robustly correlated, p &gt; 0.05. Biliary CEA values differed significantly across Duke's stages, with particular significance between the control group and Duke's stage D (p &lt; 0.001). Among patients without intraoperative liver metastasis but with biliary CEA &gt;10 ng/ml, 80% developed liver metastasis postoperatively despite undergoing chemotherapy. Conclusion: This research highlights the pivotal role of biliary CEA levels in diagnosing latent hepatic metastases in colorectal cancer. Demonstrating superior sensitivity compared to traditional serum tests, biliary CEA presents a potential shift in early detection. Elevated levels suggest expanded therapeutic options, from timely hepatectomies to adopting radiofrequency ablation and anti-CEA monoclonal antibodies, enhancing the likelihood of better patient outcomes. Essentially, these findings enhance our understanding of colorectal cancer, emphasizing the importance of personalized care, early diagnosis, and customized treatments, thereby directing us toward better patient outcomes.