Mutation patterns in colorectal cancer and their relationship with prognosis.

Background: Colorectal cancer (CRC) is a prevalent malignancy and a leading cause of cancer-related mortality. Extensive research into the aetiology of CRC has revealed that somatic mutations in certain genes play a crucial role in CRC development.AIM: In this study, we utilized data from public databases to investigate prevalent mutation patterns in CRC and developed a prognostic predictive model for CRC patients based on mutant genetic characteristics and other relevant clinical features. Methods: We initially gathered mutation information from CRC patients by analysing data from 15 datasets to identify genes with a mutation frequency of ≥10 %. Next, log-rank analyses were used to determine the relationship between prognosis and the mutational status of the most commonly mutated genes; the SIGnaling database was utilized to generate a protein‒protein interaction network. We consolidated and classified the gene mutation patterns of CRC patients in the database based on frequently mutated genes related to prognosis. A predictive nomogram was constructed, including age, sex, TNM stage, and mutation partner, based on available clinical, mutational, and prognostic information for CRC patients at our institution. Finally, the reliability of the model was verified using time-dependent ROC curve analysis. Results: The top 7 genes somatically mutated ≥10 % in 4477 samples from 4255 patients were TP53 (67 %), APC (66 %), KRAS (43 %), PIK3CA (18 %), FBXW7 (14 %), SMAD4 (14 %), and BRAF (10 %). Log-rank analysis demonstrated that the mutation status of 5 genes, namely, TP53, APC, PIK3CA, SMAD4, and BRAF, correlated significantly with prognosis. Protein‒protein interaction analysis confirmed functional interactions between these 5 genes, implicating them in tumorigenesis. We exhaustively enumerated the mutation patterns involving these five genes in 4255 patients, resulting in identification of 32 mutational patterns. After consolidation and classification, these patterns were divided into 3 grades based on patient prognosis. Next, a predictive nomogram based on the clinical, mutational, and prognostic information of 107 CRC patients treated at University Medical Center Rostock was constructed. The area under the curve (AUC) values for the model for predicting 1-, 3-, and 5-year overall survival were 0.779, 0.721, and 0.815, respectively. Conclusion: Common mutational patterns based on frequently mutated genes are associated with prognosis in CRC patients. Our study provides a valuable and concise prognostic predictor for determining outcomes in patients with CRC.

Peptide-stimulated T cells bypass immune checkpoint inhibitor resistance and eliminate autologous microsatellite instable colorectal cancer cells.

Two hypermutated colon cancer cases with patient-derived cell lines, peripheral and tumor-infiltrating T cells available were selected for detailed investigation of immunological response.T cells co-cultured with autologous tumor cells showed only low levels of pro-inflammatory cytokines and failed at tumor recognition. Similarly, treatment of co-cultures with immune checkpoint inhibitors (ICI) did not boost antitumor immune responses. Since proteinase inhibitor 9 (PI-9) was detected in tumor cells, a specific inhibitor (PI-9i) was used in addition to ICI in T cell cytotoxicity testing. However, only pre-stimulation with tumor-specific peptides (cryptic and neoantigenic) significantly increased recognition and elimination of tumor cells by T cells independently of ICI or PI-9i.We showed, that ICI resistant tumor cells can be targeted by tumor-primed T cells and also demonstrated the superiority of tumor-naïve peripheral blood T cells compared to highly exhausted tumor-infiltrating T cells. Future precision immunotherapeutic approaches should include multimodal strategies to successfully induce durable anti-tumor immune responses.

Establishment of the molecular subtypes and a risk model for stomach adenocarcinoma based on genes related to reactive oxygen species.

Background: Oxidative stress promotes the development of stomach adenocarcinoma (STAD) and resistance of STAD patients to chemotherapy. This study developed a risk classification and prognostic model for STAD based on genes related to oxidative stress. Methods: Univariate Cox regression and least absolute shrinkage and selection operator (Lasso) regression analysis were performed using transcriptome data of STAD from The Cancer Genome Atlas (TCGA) and reactive oxygen species (ROS)-related genes from Gene Set Enrichment Analysis (GSEA) website to develop a risk model. Genetic landscape, pathway characteristics and immune characteristics between the two risk groups were assessed to evaluate patients' response to anti-tumor therapy. Further, a nomogram was created to evaluate the clinical outcomes of STAD patients. The mRNA levels of genes were detected by reverse transcription quantitative PCR (RT-qPCR). Results: Two ROS-related molecular subtypes (subtype C1 and C2) were classified, with subtype C2 having unfavorable prognosis, higher immune score, and greater infiltration of macrophages, myeloid-derived suppressor cells (MDSCs), mast cells, regulatory T cells, and C-C chemokine receptor (CCR). Five ROS-related genes (ASCL2, COMP, NOX1, PEG10, and VPREB3) were screened to develop a prognostic model, the robustness of which was validated in TCGA and external cohorts. RT-qPCR analysis showed that ASCL2, COMP, NOX1, and PEG10 were upregulated, while the mRNA level of VPREB3 was downregulated in gastric cancer cells. The risk score showed a negative relation to tumor mutation burden (TMB). Low-risk patients exhibited higher mutation frequencies of TTN, SYNE1, and ARID1A, higher response rate to immunotherapy and were more sensitive to 32 traditional chemotherapeutic drugs, while high-risk patients were sensitive to 13 drugs. Calibration curve and DCA confirmed the accuracy and reliability of the nomogram. Conclusion: These findings provided novel understanding on the mechanism of ROS in STAD. The current study developed a ROS-related signature to help predict the prognosis of patients suffering from STAD and to guide personalized treatment.

Increased expression of GIPC2 in colon adenocarcinoma is associated with a favorable prognosis and high levels of immune cell infiltration.

Gα­interacting protein C­terminus PDZ­domain­containing family member 2 (GIPC2) serves an important role in the development of digestive tract tumors; however, its role in colon adenocarcinoma (COAD) has yet to be elucidated. In the present study, data were retrieved from The Cancer Genome Atlas database to investigate the association between GIPC2 expression and prognosis, as well as the levels of tumor­infiltrating immune cells. Immunohistochemical analysis was subsequently performed on 22 pairs of COAD and adjacent normal colon tissues, which were collected during surgery, to verify GIPC2 protein expression. The results showed that the positive rate in the normal intestinal mucosa group (18/22, 81.82%) was significantly higher than that in the COAD group (3/22, 13.64%, χ2=20.497, P<0.001). Gene set enrichment analysis was used to predict the signaling pathways regulated by GIPC2 in COAD, whereas the CIBERSORT algorithm was used to analyze the association between GIPC2 expression and immune cell infiltration. The expression levels of GIPC2 were revealed to be significantly downregulated in COAD compared with in normal colon tissues (P<0.05). Notably, the overall survival (P=0.004), disease­specific survival (P=0.003) and progression­free interval (P=0.011) rates of the group with high GIPC2 expression were higher compared with those in the group with low GIPC2 expression. In addition, the results of the regression analysis suggested that GIPC2 was an independent prognostic factor for COAD (P=0.007). The expression levels of GIPC2 were significantly associated with tumor stage, lymph node status and lymphatic invasion, and GIPC2 expression was enriched in 'cell cycle checkpoints', 'DNA replication' and 'mitosis­associated signaling pathways'. In addition, a positive association was observed between high GIPC2 expression and levels of infiltrating immune cells. Moreover, the expression of immune checkpoint­associated genes was significantly higher in the group with low GIPC2 expression. Taken together, the findings of the present study demonstrated that high expression levels of GIPC2 were associated with a favorable prognosis and increased infiltration of immune cells in COAD; therefore, GIPC2 may serve as a biomarker to assess prognosis and the level of immune cell infiltration in patients with COAD.

Increased ANXA5 expression in stomach adenocarcinoma infers a poor prognosis and high level of immune infiltration.

BACKGROUND: The prognostic role of annexin A5 (ANXA5) in stomach adenocarcinoma (STAD) has not been studied, and its relationship with immune infiltration is still unclear. OBJECTIVE: This investigation aimed at exploring the role of ANXA5 in STAD using an integrated bioinformatics analysis. METHODS: The expression of ANXA5 in STAD and the correlations between the effect of ANXA5 and survival of STAD patients were investigated using database. The clusterProfiler package in R software was used to perform enrichment analysis on the top 100 co-expressed genes of ANXA5 from the COXPRESdb online database. Correlations between ANXA5 and immune cell infiltrates were analyzed using the TIMER database. RESULTS: In STAD, ANXA5 expression was significantly upregulated and increased ANXA5 expression was significantly correlated with poor overall survival (P< 0.05). In multivariate analysis, upregulated ANXA5 expression was an independent predictive factors of poor prognosis (P< 0.05). The co-expressed genes were involved in extracellular matrix (ECM)-related processes. In STAD, ANXA5 expression was significantly correlated with various infiltrating immune cells (P< 0.05). CONCLUSIONS: Together with our findings, ANXA5 could serve as a potential biomarker to assess prognosis and immune infiltration level in STAD.

Pathological complete response from oral chemotherapy combined with trans-arterial chemotherapy and embolization in an unresectable gastric cancer patient: A case report.

RATIONALE: Gastric cancer is still one of the most common cancer in East Asia. More than 70% gastric cancer patients are diagnosed at an advanced stage in China. Moreover, about 10% cases are unresectable which usually suffer a poor prognosis with a median survival time of 5 to 12 months. In recent years, some clinical studies found that many unresectable gastric cancer cases could get opportunity for surgery after treatment that improve prognosis significantly PATIENT CONCERNS:: 64-year-old male patient was admitted with upper abdominal pain. Upper gastrointestinal endoscopy showed a large ulcerated tumor located from the cardia to the anterior wall of the upper gastric body. Histopathological examination showed it was moderately differentiated adenocarcinoma. Computed tomography (CT) scan image showed a large bulging mass with internal ulcer at the lesser curvature wall, left gastric artery and coeliac trunk were surrounded by fused lymph nodes. DIAGNOSES: Based on the histopathological examination and imaging findings, patient was diagnosed advanced gastric cancer and hardly to resect radically. INTERVENTION: Oral chemotherapy combined with trans-arterial chemotherapy and embolization (TACE) was initiated. Eight weeks after initial therapy, radical laparoscopy-assisted total gastrectomy with D2 lymph node dissection and Roux-en-Y anastomosis were performed successfully. OUTCOMES: Patient was discharged on postoperative day 11 without complications. Histological analysis of the specimen and resected 31 lymph nodes revealed no malignancy. The patient experienced a pathological complete response (pCR). LESSONS: In this case, oral chemotherapy combined with TACE which was rarely reported in the treatment of unresectable gastric cancer achieves a great therapeutic benefit. Although further clinical studies will be needed to establish, it may be a potent strategy for degrading stage and supplying a new chance for surgery.

LncRNA H19 functions as a competing endogenous RNA to regulate AQP3 expression by sponging miR-874 in the intestinal barrier.

Intestinal barrier dysfunction is a significant clinical problem, which develops in a variety of acute or chronic pathological conditions. In a previous study, we found that microRNA-874 (miR-874) suppresses aquaporin-3 (AQP3) expression, which contributes significantly to intestinal barrier dysfunction. Recently, a new regulatory circuit was identified in which RNA can crosstalk with each other by competing for shared miRNA. Here, we show that the human long noncoding RNA (lncRNA) H19 may function as a competing endogenous RNA (ceRNA) to regulate the expression of AQP3 through competition for miR-874, thus playing a significant role in maintaining intestinal barrier function.

Laparoscopy-Assisted D2 Radical Gastrectomy for Advanced Gastric Cancer.

BACKGROUND/AIMS: The aim of this study was to investigate the efficacy and advantages of laparoscopy-assisted gastrectomy (LAG) with D2 lymphadenectomy versus conventional open D2 gastrectomy (ODG) in advanced gastric cancer (AGC) patients. METHODOLOGY: From June 2009 to June 2014, 233 patients who were treated by conventional radical ODG and 188 cases who underwent radical LAG for AGC at our department were enrolled in this study. Clinical data recorded in hospital database was retrospectively reviewed and analyzed. RESULTS: There were no significant differences in age, gender or preoperative laboratory tests distribution between the LAG group and the ODG group. Two (1%) of the patients who underwent LAG required conversion to ODG. The advantages of minimally invasive surgery including earlier recovery of bowel movements and shorter postoperative hospitalization time were observed in LAG group. The number of harvested lymph nodes was 24.3 ± 3.3 in the LAG group and 25.0 ± 2.8 in ODG (p = 0.110). CONCLUSIONS: Although prospective randomized trials with long follow-up period are needed to identify the feasibility, we have shown the safety and advantages of LAG with D2 lymphadenectomy for treating AGC patients in this study.

Sentinel lymph node and skip metastases in gastric cancer: a prospective study.

BACKGROUND/AIMS: The aim of the present study was to investigate localization and distribution of sentinel lymph node (SLN) in gastric cancer and influenced factors of skip metastasis. METHODOLOGY: Eighty-nine cases with solitary or single station metastatic lymph nodes (LNs) among 779 gastric cancer patients were included in this study. D2 or D2+ lymph node dissection with total or distal gastrectomy had been performed. The location of SLNs and characteristics were analyzed retrospectively. RESULTS: Seventy-one patients (79.8%) had SLNs at the first level of the regional LN group and 18 cases (20.2%) had skip metastases SLNs. NO.7 (44.4%) and NO.8a (27.8%) were the most common sites of skip metastases. Multivariate analysis identified the differentiated degree of tumor as the only significant influenced factor of skip metastases (p=0.031). The number of metastatic SLNS was significantly greater in advanced gastric cancer patients (2.4 ± 1.4) than early gastric cancer patients (1.5 ± 0.9) (pT4 vs. pT1, p=0.002; pT4 vs. pT2, p=0.014; pT3 vs. pT1, p=0.018). CONCLUSIONS: The differentiated degree of gastric cancer is the most important influenced factor of skip metastases and NO.7 and NO.8a are the most important N2 LN stations that should be observed when lymphadenectomy is performed.

Beneficial effects of S-adenosyl-L-methionine on post-hepatectomy residual liver function: a prospective, randomized, controlled clinical trial.

BACKGROUND/AIMS: Hepatectomy is associated with high rates of postoperative liver dysfunction in patients with cirrhosis. Since S-adenosyl-L-methionine (SAMe) can be used to treat liver disease, we performed a prospective clinical trial to investigate whether it could be used after hepatectomy to benefit residual liver function. METHODOLOGY: We studied 79 hepatitis-related chronic patients who underwent resection of hepatocellular carcinoma; 39 patients were randomly assigned to receive postoperative intravenous SAMe treatment, and 40 were randomly assigned to a control group. The postoperative SAMe treatment consisted of SAMe 1,000mg given intravenously for seven days. The other treatment was standardized. RESULTS: At inclusion into the trial no significant differences were observed between the two groups with respect to gender, age, Child classification, preoperative liver function tests, blood loss, total time of hepatic pedicle occlusion and the extent of liver resection. The overall frequency of postoperative liver insufficiency decreased from 42% in the control group to 31% in the SAMe group, although not statistically significant (p=0.121). When the patients who underwent hepatic pedicle occlusion by Pringle's maneuver over 15min were analyzed, the frequency of postoperative liver insufficiency (p=0.028), serum total bilirubin levels on days 5 (p=0.025) and 7 (p=0.032) preoperatively, and the maximum value of postoperative serum total bilirubin (p=0.040) were significantly greater in the control than in the SAMe group. CONCLUSIONS: The results indicate that the postoperative SAMe therapy can benefit residual liver function of the patients with cirrhosis, especially in those suffering greater ischemia reperfusion injury.