BACKGROUND:Some studies show that cytoreductive surgery with hyperthermic intraperitoneal chemotherapy (HIPEC) may improve overall survival and is a possible curative treatment for selected colorectal cancer (CRC) patients with restricted peritoneal metastasis (PM). The value of HIPEC in preventing PM of CRC is still controversial. MATERIALS AND METHODS:In this retrospective propensity score matching (PSM) cohort study, all patients with cT4N0-2M0 undergoing treatment at a single institution in China (2014-2018) were reviewed. The 3-year disease-free survival (DFS) was set as the primary outcome, and the 3-year PM rate was also analyzed. RESULTS:220 patients were included in this study for analysis. After 1:3 PSM: HIPEC (n = 45) and No HIPEC (n = 135). Through analysis, it was found that prophylactic HIPEC correlated to better DFS [hazard ratio (HR) 0.43, 95 % confidence interval (CI) 0.19-0.95; p = 0.037], and N2 stage correlated to worse DFS [HR 1.97, 95 % CI 1.09-3.56; p = 0.025]. For laparoscopic surgery subgroup analyses, 3-year PM rate of patients with laparoscopic surgery was 13.8 % in No HIPEC group, and 2.6 % in HIPEC group (p = 0.070). Besides, no post-operative death occurred, the anastomotic leakage rate was 2.2 % in HIPEC group and 0.7 % in the control group (p = 0.439). CONCLUSIONS:Prophylactic HIPEC may improve the prognosis in patients with cT4N0-1M0 CRC, but not in cT4N2M0 CRC, and it does not significantly increase surgery-related complications. Laparoscopic surgery followed by HIPEC for T4 stage CRC may not increase risk of PM.

Objective: This study (CRD42023464989) aimed to explore the effects of pre-operation immunonutrition on safety and immune related factors in colorectal cancer patients undergoing surgery. Methods: We systematically searched PubMed, Embase, and Wanfang databases to collect all clinical randomized controlled trials of the application of pre-operation immunonutrition for patients with colorectal cancer, published until July 2023. The primary outcomes were safety and immune related factors. Results: A total of 16 studies were finally included. Preoperative immunonutrition could reduce the postoperative infection rate (risk ratio (RR) = 0.56, 95% confidence interval (CI): 0.36, 0.88; p = .01), and wound infection rate (RR = 0.44, 95% CI: 0.27, 0.70; p < .001) in patients with colorectal cancer. For length of stay (mean difference (MD) = -1.10, 95% CI: -2.70, 0.49; p = .17), it was similar between groups. Meanwhile, patients in the pre-operation immune nutrition group also had significantly increased infiltrative lymphocytes CD16+ (MD = 0.04, 95% CI: 0.02, 0.06; p < .001), and CD56+ (MD = 0.05, 95% CI: 0.03, 0.06; p < .001) cells in the tumor tissues, compared to the control group. Conclusion: Immunonutrition intervention has the potential to reduce postoperative infectious complications and improve tumor infiltrative lymphocytes in patients with colorectal cancer undergoing surgery.

Investigating the potential of macrophage efferocytosis has led to a growing interest in exploiting macrophage in immunotherapy. Macrophage-specific nano-medicines are currently being developed to stimulate the phagocytic clearance of apoptotic debris in atherosclerosis; however, these unnatural nanoparticles exhibit poor biocompatibility and limited efferocytosis activation ability. Here a pro-efferocytosis biomimetic nanorobot UM-EVLipo is developed based on nano-motor and extracellular vesicles (EVs) hybridized with plaque double-targeted liposomes. An elegant procedure with only 4 steps is integrated to prepare and purify these hybrid nanovesicles. It is first demonstrated that the EVs derived from bone marrow macrophages stimulated with IL-4 can promote the transformation of M0 and M1 macrophages into M2, an efferocytosis phenotype. Furthermore, the EVs hybridized with SHP-1 siRNA-liposomes blocked the "don't eat me" signal from apoptotic debris by intervening CD47-SIRP alpha-SHP-1 axis. This two-pronged strategy of resetting phenotype and blocking checkpoints furthest activates macrophage efferocytosis. Importantly, the urease motor increased the swimming speed of nanovesicles in the blood by 12.4 times, making it easier for them to penetrate the endothelial barrier. The UM-EVLipo accumulated within the atherosclerotic plaque, reactivate lesioned efferocytosis and reduce the plaque area in the carotid and coronary, which demonstrates the potential of a two-pronged strategy to prevent atherosclerotic cardiovascular disease.

The previous activatable probe toward one analyte can still produce imprecise signal as a result of the challenge cased by accurate identification in complex biological environment. As smarter molecules, dual-locked probes are able to respond to two different analyte with one or more output signals, which display significantly reduced background interference and increased spatial resolution. In this study, a smart cysteine/lipid droplets sequenceactivated dual-locked fluorescent probe CN-NO2 has been designed and synthesized for the detection of tumor tissues. The recognition group of CN-NO2 removed after reacting with cysteine (Cys) and undergoing intramolecular rearrangement to generate complete fluorophores. This structure had a strong solvent effect; it could recognize lipid droplets (LDs) in cells, thus exhibiting fluorescence without secondary molecular adjustment. The fluorescence of CN-NO2 was enhanced 1752-fold after double unlocking. Importantly, the unlocking process only takes 10 min. Furthermore, the fluorescence of CN-NO2 was amplified remarkable and anchored at LDs after reacting with Cys in A549 cells. Therefore, CN-NO2 clearly indicated the site of mouse xenograft tumor in vivo. Such sequence-activated dual-locked fluorescent probes are envisioned to execute more analysis and diagnostic tasks giving these unique characteristics.

Although the significance of the curative effect has been recognized, ideal nanocarriers with the properties of lysosomal escape and biocompatibility are still lacking for the development of siRNA-based cancer gene therapy. In this work, a lysosomal escaped and redox-responsive polyacrylamide nanohydrogel (cRGD-9R-PA(ss)) was constructed for STAT3 siRNA delivery. The functional groups, including tumor-targeting peptide cRGD, cell-penetrating and lysosome-escaping peptide 9R, and a redox-responsive disulfide bond, were introduced to the polyacrylamide nanohydrogel to attain enhanced transfection efficiency and biocompatibility. The synthesized cRGD-9R-PA(ss)(siRNA) was dispersed as core-shell nanoparticles in water with an average size of 48 nm. cRGD-9R-PA(ss)(siRNA) could prevent lysosomal phagocytosis and responsive release STAT3 siRNA into C26 tumor cells, thus promoting STAT3 gene silencing and inhibiting the proliferation of cancer cells in vitro and in vivo. Our results demonstrate that cRGD-9R-PA(ss) hydrogel nanospheres constitute a potential candidate vector for siRNA-based colon cancer gene therapy.

Colorectal cancer (CRC) has become a global health problem which has almost highest morbidity and mortality in all types of cancers. This study aimed to uncover the biological functions and underlying mechanism of MCM8 in the development and progression of CRC. The expression level of MCM8 was found to be upregulated in CRC tissues and significantly associated with tumor grade and patients' survival. Knocking down MCM8 expression in CRC cells could restrain cell growth and cell motility while promoting cell apoptosis in vitro, as well as inhibit tumor growth in xenograft mice model. Based on the RNA screening performing on CRC cells with or without MCM8 knockdown and the following IPA analysis, CHSY1 was identified as a potential target of MCM8 in CRC, whose expression was also found to be higher in tumor tissues than in normal tissues. Moreover, it was demonstrated that MCM8 may regulate the expression of CHSY1 through affecting its NEDD4-mediated ubiquitination, both of which synergistically execute tumor promotion effects on CRC. In conclusion, the outcomes of our study showed the first evidence that MCM8 act as a tumor promotor in CRC, and may be a promising therapeutic target of CRC treatment.

As one of the most common malignancies, colorectal cancer (CRC) requires a thorough understanding of the mechanisms that promote its development and the discovery of new therapeutic targets. In this study, immunohistochemical staining confirmed significantly higher expression levels of KIF15 in CRC. qPCR and western blot results demonstrated the effective suppression of KIF15 mRNA and protein expression by shKIF15. Downregulation of KIF15 inhibited the proliferation and migration of CRC cells while promoting apoptosis. In addition, evidence from the xenograft experiments in nude mice demonstrated that KIF15 knockdown also suppressed tumor growth. Through bioinformatics analysis, the downstream molecular NRAS and Rac signaling pathway associated with KIF15 were identified. KIF15 knockdown was found to inhibit NRAS expression and disrupt Rac signaling pathway. Moreover, WB and Co-IP assays revealed that KIF15 reduced the ubiquitination modification of NRAS protein by interacting with the E3 ligase MDM2, thereby enhancing NRAS protein stability. Functionally, NRAS knockdown was shown to inhibit cell proliferation and migration. In conclusion, KIF15 promoted CRC progression by regulating NRAS expression and Rac signaling pathway.

Purpose In recent years, natural orifice specimen extraction surgery (NOSES) has gained widespread attention as an alternative approach. Although the safety and feasibility of NOSES have been well documented, many questions remain open for discussion. The aim of this guideline is to provide more evidence for the promotion of NOSES. Methods This guideline has been prepared by the CACA Committee of Colorectal Cancer Society and the International NOSES Alliance, based on the latest evidence. Results The guideline on NOSES for colorectal cancer include the definition, classification, technology requirement, indications, technical difficulties and clinical research. Conclusion The guideline provides a full introduction of the theoretical and technical aspects of NOSES for colorectal cancer which will beneficial to development of NOSES.

Objective To systematically evaluate the efficacy and safety of anlotinib for patients with metastatic colorectal cancer (mCRC). Method We searched eligible studies in PubMed, Embase, Cochrane Library, Wanfang Patent Database and China National Knowledge Infrastructure (CNKI) from database inception to September 3, 2022. A total of four random control studies were included to extract relevant data, including objective response rate (ORR), disease control rate (DCR) and the rate of adverse events. The Stata version 14.0 software was used to calculate the risk ratio (RR) with 95% confidence intervals (95% CI). Results This meta-analysis included 618 patients from four clinical studies. The pooled RR for ORR and DCR were 1.54 (95% CI: 1.08–2.20, P＜ 0.05) and 1.38 (95% CI: 1.04–1.81, P＜ 0.05), indicating that anlotinib administration significantly improved ORR and DCR in patients with mCRC. The pooled RR for the incidence of diarrhea was 1.64 (95% CI: 0.72–3.73, P = 0.24) and the rate of leukocytopenia was 2.32 (95% CI: 1.16–4.62, P＜ 0.05). Conclusion Anlotibin, an oral multi small-molecule receptor tyrosine kinase inhibitor, may provide clinical benefits for patients with mCRC in terms of ORR and DCR. Adverse events associated with anlotinib were tolerable.

The objective of this study was to summarize relevant data from previous reports and perform a meta-analysis to compare short-term surgical outcomes and long-term oncological outcomes between emergency and elective surgery for colorectal cancer (CRC). A systematic literature search was performed using PubMed and Embase databases, and relevant data were extracted. Postoperative morbidity, hospital mortality within 30 days, postoperative recovery, overall survival (OS), and relapse-free survival (RFS) were compared using a fixed or random-effect model. A total of 28 studies involving 353,686 participants were enrolled for this systematic review and meta-analysis, and 23.5

Abstract The phenomenon of tumor hierarchy and genetic instability can be explained by the “two‐hits theory” and results in the occurrence of many somatic mutations. The expression of nonsynonymous mutations results in the production of mutant proteins from tumor cells, namely tumor‐specific antigens called neoantigens. Because neoantigens do not exist in healthy cells, they have the potential to stimulate antitumor immune responses by CD4+ and CD8+ T‐cell activation without jeopardizing normal tissues. Immunotherapy has reshaped the cancer treatment paradigm in recent decades with the introduction of immune‐checkpoint blockade therapy and transgenic T‐cell receptor/chimeric antigen receptor T cells. However, these strategies performed poorly in solid tumors because of the obstacles of the immunosuppressive microenvironment caused by regulatory T cells and other suppressor cells. Therefore, other immunotherapeutic strategies are under development, such as personalized vaccines, to trigger de novo T‐cell responses against neoantigens and lead to the amplification of tumor‐specific T‐cell subclones. Neoantigen epitope prediction algorithms have enabled the detection of neoantigens and the creation of tailored neoantigen vaccines as a result of the fast development of next‐generation sequencing and cancer bioinformatics. Here we provide an overview of the current neoantigen cancer vaccines and adoptive T‐cell transfer therapy with neoantigen‐specific lymphocytes. We also discuss the challenges in developing neoantigen‐targeted immunotherapeutic strategies for cancer.

With the rapid development of minimally invasive surgery for the treatment of colorectal diseases,novel surgical techniques aim-ing at reducing surgical trauma have become frequently discussed issues[1].Natural orifice specimen extraction surgery(NOSES),including transvaginal specimen extraction(TVSE)and transanal specimen extraction(TASE),have been considered well-estab-lished surgical approaches for avoiding abdominal incision[2,3].NOSES represents one of the least invasive options in the treatment of colorectal cancer(CRC),but most relevant studies are single-centered with limited sample sizes.Whether previous findings can be generalized to a wider patient population remains unclear[3,4].Therefore,we performed the first nationwide large-scale study with the aim of confirming the safety and feasibility of NOSES for CRC by analyzing the short-term and long-term oncolog-ical outcomes.

Background Gastric cancer (GC), the most commonly diagnosed cancer worldwide with poor 5-year survival rate in advanced stages. Although immune-related and survival-related biomarkers, which typically comprise aberrantly expressed long non-coding RNAs (lncRNAs) and genes, have been identified, there are no reports of immune-related lncRNA pair (IRLP) signatures for GC. Methods In this study, we acquired lncRNA expression profiles from The Cancer Genome Atlas (TCGA) and used the least absolute shrinkage and selection operator (LASSO) Cox proportional hazards model (iteration = 1000) to develop a IRLP prognostic signature. The area under curve (AUC) was used to assess the prognosis predictive power. The multivariate Cox regression analysis was performed to identify whether this signature was an independent prognostic factor. The immune cell infiltration analysis was performed between the two risk groups. Last, molecular experiments were performed to explore LINC01082 is involved in the development of GC. Results We acquired lncRNA expression profiles and used the LASSO Cox model to develop an 18-IRLP signature with a strong prognostic predictive power. The 5-year AUC values of the training, validation, and overall TCGA datasets were 0.77, 0.86, and 0.80, respectively. The different prognostic outcomes between the high- and low-risk groups were determined using our 18-IRLP signature. Moreover, our 18-IRLP signature was an independent prognostic factor as per the multivariate Cox regression analysis, and showed better prognostic evaluation than the traditional TNM staging system as well as other clinical features. We also found differences in cancer-associated fibroblast and macrophage M2 infiltration and the expression of PD-L1, CTLA4, LAG3, and HLA were also observed between the two risk groups (P < 0.05). Analysis of biological functions revealed that target genes of the lncRNAs in the IRLP signature were enriched in focal adhesion and regulation of actin cytoskeleton. Finally, as one of significant candidates of IRLP signature, overexpression of LINC01082 suppressed the invasion ability of GC cells as well as PD-L1 expression profiles. Conclusions Our novel 18-IRLP signature provides new insights regarding immunological biomarkers, imparts a better understanding of the tumor immune microenvironment, and can be used for predicting prognosis and evaluating immune response in GC.

Colorectal cancer (CRC) is the second leading cause of cancer deaths in developed countries [1]. The malignant transformation from small clumps to cancer takes about 10 years [2]. This study aimed to characterize proteomic dynamics associated with CRC development and progression, and identify novel therapeutic targets for intercepting the underlying oncogenic processes. We have optimized pressure cycling technology (PCT) coupled with data-independent acquisition mass spectrometry (DIA-MS) for robust and reproducible proteomic analysis of biopsy-level formalin-fixed paraffin-embedded (FFPE) tissues [3]. In this study, we profiled the proteomic tissue landscape of CRC evolving from normal colon to hyperplastic polyps, adenomas, adenocarcinoma not otherwise specified (AC) or mucinous adenocarcinoma (MC). We identified 69,949 peptides, 6,359 protein groups, and 4,830 unique proteins (Supplementary Table S1) based on our previously established spectral library for DIA analysis [4] from 170 FFPE tissue samples (85 patients, each with 2 biological replicates) (Figure 1A). Pearson's correlation coefficient between biological replicates was 0.813, and 0.953 between technical replicates. We identified 928 differentially expressed proteins by comparing protein expression in samples from different CRC clinical stages to normal colon tissue samples (Figure 1B). Pairwise comparisons between polyps and normal colon, adenomas and polyps, carcinoma and adenomas, as well as MC and AC revealed distinct proteomic changes associated with each transformation towards malignancy (Supplementary Figure S1A). Canonical pathways analysis revealed that the dysregulated proteins were mostly related to oxidative phosphorylation. Interestingly, oxidative phosphorylation was enhanced in precancerous tissues (hyperplastic polyps and adenomas) but suppressed in CRC tissues, suggesting metabolic adaptations of tumor cells in the evolving microenvironment (Supplementary Figure S1B). Analysis of diseases and biological functions of differential proteins in benign lesions showed proteomic perturbations associated with oncogenic pathways. For example, COPE, COPA, and COPZ1 are proteins encoded by coatomer protein complex genes which are essential proteins for tumorigenesis in CRC. PSMC3, PSMD13, PSMA7 and PSMD8 are all proteasomal proteins whose expressions began to rise in polyps and peaked in adenomas (Supplementary Figure S1C). Six biologically significant protein expression patterns associated with CRC development were selected by unsupervised cluster analysis. Patterns 1, 2 and 3 were formed by upregulated proteins while patterns 4, 5 and 6 were formed by downregulated proteins (Figure 1C). Gene Ontology analysis for enrichment of biological processes in the six clusters (Supplementary Figure S2A) identified extracellular matrix (ECM) enrichment in pattern 3. We then checked for all ECM-related proteins in the "matrisome", which has been defined as the combination of core ECM proteins (glycoproteins, collagens, and proteoglycans) and ECM-associated proteins (ECM-affiliated proteins, ECM regulators, and secreted factors) [5]. Among the six protein expression patterns, we observed enrichment of ECM regulators in the upregulated patterns. Proteins within each pattern formed protein-protein interaction networks using Cytoscape with the GeneMania plugin (Supplementary Figure S2B). We then narrowed our focus on proteins that were consistently upregulated along the stages of tumor progression. Among plasma membrane, nucleus, cytoplasm, extracellular space and other locations, our data showed that the cytoplasm and extracellular proteins stood out as the locations with the highest expression of dysregulated proteins (Supplementary Figure S2C). CRC progression was associated with substantially increased expression of multiple enzymes (Supplementary Figure S3A). Of note, Procollagen-Lysine, 2-Oxoglutarate 5-Dioxygenase 2 (PLOD2) was the most up-regulated protein (Figure 1D and Supplementary Figure S3B). We further performed a pairwise comparison between pre-cancerous and cancerous samples with benign samples, and PLOD2 consistently outstood as the top hit (Supplementary Figure S3C). Next, we randomly selected eight samples of each tissue type (normal colon, hyperplastic polyps, adenomas, AC, and MC) for targeted measurement of PLOD2 using parallel reaction monitoring (PRM). The PRM data from the 40 samples confirmed the elevation of PLOD2 (Figure 1E). As further verification, western blot (WB) analysis was performed on four new CRC patients and observed higher PLOD2 expression in CRC tissues than in matched para-tumoral normal colon (Figure 1F). We also assessed PLOD2 expression by immunohistochemistry staining (IHC) of tissue microarrays (TMAs) containing 118 CRC (8th AJCC TNM Stage II) and 79 para-tumoral normal colon tissues (Supplementary Table S2). The IHC staining of PLOD2 in para-tumoral normal colon tissues showed that 68 samples had < 25% positive colon cells, eight samples had 25%-50%, and only three samples had > 50%. In contrast, CRC tissues showed significantly higher PLOD2 expression than para-tumoral normal colon (Figure 1G). Remarkably, we found that higher PLOD2 expression was associated with poorer overall survival of CRC patients (Figure 1H, Supplementary Table S2). Next, we measured the PLOD2 expression in six CRC cell lines and chose the two with the highest PLOD2 expression, namely HCT116 and HT-29, for generating PLOD2-knockout (KO) congener lines using CRISPR-Cas9. Each congenic pair of cell lines was treated with increasing concentrations of minoxidil, a lysyl hydroxylase inhibitor of PLOD2. Minoxidil inhibited PLOD2 expression in both wild-type cell lines in a time- and dose-dependent manner, while clonogenicity and cell proliferation were dramatically suppressed when PLOD2 was inhibited by minoxidil or knocked-out (Figure 1I). In addition, both minoxidil treatment and PLOD2-KO suppressed CRC cell migration and invasion (Figure 1J). Extending our findings to in vivo model, we injected HCT116 and HT-29 cell lines subcutaneously into nude mice. We tested the effects of placebo versus minoxidil treatment in vivo on tumors generated by wild-type HCT116 and HT-29 cells, and also compared the growth of tumors generated by PLOD2-KO HCT116 and HT-29 cells with PLOD2-high tumors. Our data showed that both minoxidil and CRISPR-Cas9-mediated PLOD2 suppression led to a significant decrease in tumor volume (Figure 1K). A second in vivo model was patient-derived xenografts (PDX) tumors from four CRC patients. Patient tumors with high PLOD2 levels (PLOD2 positive tumor cells were > 80%) were sensitive to minoxidil inhibition while PLOD2-negative tumors were resistant (Figure 1L), highlighting the potential clinical application of targeted therapy against PLOD2. To gain mechanistic insight on how PLOD2 inhibition suppresses CRC tumors, we compared the transcriptome and proteome of HCT116-KO and the HCT116-normal control (NC) cell lines using RNA sequencing and DIA-MS, respectively (Supplementary Table S3). We identified 1236 up- and 955 down-regulated transcripts, and 227 up- and 127 down-regulated proteins (Supplementary Figure S4A). The data indicated that PLOD2 contributed to tumor growth, resistance to cell necrosis, and was closely related to the development of colorectal cancer (Supplementary Figure S4B). PLOD2 was also involved in protein synthesis, metabolism, and mRNA translation (Supplementary Figure S4C). Selected protein networks prioritized by these analyses are shown in Supplementary Figure S4D and E. An overview of the patients' basic pathological characteristics is shown in Supplementary Table S4. Compared to previous studies [6, 7], our study systematically tracked a plethora of protein changes in CRC tissues as the disease progressed through increasing degrees of malignancy. Therapeutic interventions directed at cancer-derived ECM and their regulatory factors may be clinically effective [8]. PLOD family proteins catalyze post-translational modifications of collagen by converting lysine to hydroxylysine, which promotes stable interactions and deposition of collagen [9]. PLOD2 could be induced in L1CAM-overexpressing CRC cell lines and promoted L1CAM-mediated CRC progression by inducing ezrin signaling and the SMAD2/3 pathway [10]. Our data collectively constitute plausible evidence for suggesting further research on PLOD2 as a promising therapeutic target in CRC tumors in the emerging practice of precision oncology. Written informed consent was obtained from all participants based on the guidelines of the Declaration of Helsinki. Human tissue samples were collected with the approval of the Institutional Ethics Committee of the Second Affiliated Hospital of Zhejiang University, School of Medicine (Zhejiang, P. R. China, No.2020-322). Animal studies and formalin-fixed paraffin-embedded samples collections were approved by the Institutional Ethics Committee of the Second Affiliated Hospital Zhejiang University School of Medicine (SYXK2018-0012). All authors read and approved the final manuscript for publication. The MS proteomics data are available on the iProX database with the project ID: IPX0001414000 and the subproject ID: IPX000141400. The raw sequence data have been uploaded to SRA with an ID: PRJNA598559. The authors declare that they have no competing interests. This work was supported by the National Key Research and Development Program of China (Grant No. 2017YFC0908200), National Natural Science Foundation of China (Grant No. 81972270, 81972492, 32027801, 21904107), the Zhejiang Provincial Science Foundation for Distinguished Young Scholars (Grant No. LR19C050001), Hangzhou Agriculture and Society Advancement Program (Grant No. 20190101A04) and 2019 Zhejiang University Academic Award for Outstanding Doctoral Candidates to YK.S (Grant No. 2019071). YKS, KLX, XZ, BTZ, XLZ, LW, YTS, DL performed experiments and data interpretation. TC, JW, SJY, LFS, XMX, LNQ, JNC, WXH, XYW, XPX, JFL, LRC, JMS, SZ provided key biological samples and materials. YKS, KLX, LW, DL, SZD, HHG, GR, YTS, WL, XC, TSZ, YZ, ZYH, TNG performed data analysis. YKS, KLX, XZ, BTZ, YTS, SZ, JMS, TNG designed the study, interpreted the data, and wrote the manuscript. All authors read and approved the final manuscript. We particularly acknowledge the Biobank of the Second Affiliated Hospital of Zhejiang University School of Medicine. We thank Westlake University Supercomputer Center for assistance in data storage and computation and the Biomedical Core facility for mass spectrometry analysis. We thank Prof. Yongzhan Nie (Air Force Medical University) for his guidance in writing. We thank Dr. Oi Lian Kon for editing the manuscript. Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.

PURPOSE:This study aimed to analyze clinicopathological, survival, prognostic factors, as well as the timing of brain metastases (BM) in colorectal cancer (CRC) using data from a Chinese center.PATIENTS AND METHODS:Data of 65 consecutive CRC patients with BM were collected from a single institution in China. The time from primary tumor surgery to the occurrence of BM was calculated. Kaplan-Meier analysis was used to evaluate cumulative survival of patients. Factors associated with prognosis of overall survival (OS) were explored using Cox's proportional hazard regression models.RESULTS:The median time interval from CRC surgery to the diagnosis of BM was 24 months. After diagnosis of BM, median OS values for patients were 11 months. Extracranial metastases occurred in 45 cases (69.2%) when BM was diagnosed, and 58.5% of these patients with lung metastases Time of BMs (P=0.018), presence of extracranial metastases (P=0.033), treatment (P=0.003), CA199 (P=0.034), CA125 (P<0.001), CA242 (P=0.018), and CA211 (P=0.012) were associated with OS of patients through univariate analysis. Multivariate analysis using a Cox regression model showed that only treatment was an independent predictor for OS (conservative treatment; HR=1.861, 95% CI=1.077-3.441; P=0.048).CONCLUSION:Surgical treatment of metastatic lesions may be an alternative choice for CRC patients with BM. Identifying the timing of brain metastases can help to detect this disease early, leading to a better survival outcome.

Background: TET1, a DNA demethylase, is encoded by TET1 gene. At present, TET1 mutation has been reported to be associated with enhanced tumor immunogenicity and activated anti-tumor immunity across multiple cancers. However, the clinical effect of TET1 mutation on immunotherapy in colorectal cancer is little known. Methods: 876 patients with colorectal cancer were enrolled in this study. Genomic profiling of DNA was performed on formalin-fixed paraffin-embedded tumor samples by NGS with 733 cancer-related genes panel. The somatic and germline mutation data were both obtained. Benign and likely benign mutations were excluded from our analysis. Programmed death ligand-1(PD-L1) expression status was determined by immunohistochemistry, evaluated with the combined positive score (CPS) and tumor proportion score (TPS). The efficiency of immunotherapy was analyzed in a cohort of 85 patients with colorectal cancer from Morris et al. Results: 40 of 876 patients (4.75%) harbored TET1 mutations, including nonsynonymous, frameshift and stopgain mutations. Totally, 66 TET1 single nucleotide mutation were identified. Median tumor mutation burden (TMB) value with TET1 mutations was 109.2 mutations/MB, significantly higher than 6.1 mutations/MB in TET1 wild-type group (p Conclusions: TET1 mutation was strongly associated with higher TMB, MSI-H and PD-L1 expression and improved OS in patients receiving immune checkpoint blockade treatment, suggesting that TET1 mutation is a novel predictive biomarker for immunotherapy in colorectal cancer. Citation Format: Shaojun Yu, Ding Zhang, Shiqing Chen. TET1 mutation as potential biomarker for immune checkpoint blockade in colorectal cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 347.

Background: Early screening for colorectal cancer (CRC) is essential to improve its prognosis. Liquid biopsies are increasingly being considered for diagnosing cancer due to low invasiveness and high reproducibility. In addition, circulating extracellular vesicles (crEVs, extracellular vesicles isolated from plasma) expressing tumour-specific proteins are potential biomarkers for various cancers. Here, we present a data-independent acquisition (DIA)-mass spectrometry (MS)-based diagnostic method for liquid biopsies. Methods: Extracellular vesicles (EVs) were isolated from culture supernatants of human CRC cell lines, and plasma of patients with CRC at different tumour stages, by overnight ultracentrifugation coupled with sucrose density gradient centrifugation. Tumour-specific EV proteins were prioritized using Tandem Mass Tag (TMT)-based shotgun proteomics and phosphoproteomics. The results were verified in a second independent cohort and a mouse tumour-bearing model using Western blotting (WB). The candidate biomarkers were further validated in a third cohort by DIA-MS. Finally, the DIA-MS methodology was accelerated to permit high-throughput detection of EV biomarkers in another independent cohort of patients with CRC and healthy controls. Results: High levels of total and phosphorylated fibronectin 1 (FN1) in crEVs, haptoglobin (HP), S100A9 and fibrinogen α chain (FGA) were significantly associated with cancer progression. FGA was the most dominant biomarker candidate. Analysis of the human CRC cell lines and the mouse model indicated that FGA+ crEVs were likely released by CRC cells. Furthermore, fast DIA-MS and parallel reaction monitoring (PRM)-MS both confirmed that FGA+ crEVs could distinguish colon adenoma with an area of curve (AUC) in the receiver operating characteristic (ROC) curve of 0.949 and patients with CRC (AUC of ROC is 1.000) from healthy individuals. The performance outperformed conventional tumour biomarkers. The DIA-MS quantification of FGA+ crEVs among three groups agreed with that from PRM-MS. Conclusion: DIA-MS detection of FGA+ crEVs is a potential rapid and non-invasive screening tool to identify early stage CRC. Abbreviations: FGA: fibrinogen α chain; CRC: colorectal cancer; crEVs: circulating extracellular vesicles; EV: extracellular vesicles;MS: mass spectrometry; WB: Western blotting; ROC: receiver operating characteristic; PRM: Parallel Reaction Monitoring; GPC1: Glypican-1; GO: Gene ontology; TEM: transmission electron microscopy; FN1: Fibronectin 1; HP: haptoglobin; TMT: Tandem Mass Tag; LC-MS/MS: liquid chromatography coupled to tandem mass spectrometry; DIA: data-independent acquisition; DDA: data-dependent acquisition; CiRT: Common internal Retention Time standards;AGC: Automatic gain control; AUC: area under curve.

Purpose: To analysis factors prognostic for peritoneal metastases (PM) from colorectal cancer (CRC) treated with surgery using data from two sources and investigate the origin and effective treatment of ovarian metastases (OM). Patients and Methods: Data from CRC patients with PM who had undergone surgery were collected from the Surveillance, Epidemiology, and End Results (SEER) database (n = 639) and a single Chinese institution (n = 60). Cumulative survival was evaluated by Kaplan-Meier analysis. Factors associated with overall survival (OS) and progression-free survival (PFS) prognosis were assessed using Cox's proportional hazard regression models. Results: Median OS values for patients who underwent surgery were 19 and 32 months in the SEER database and Chinese center, respectively. Age was an independent predictor of OS in both datasets. Signet-ring cell cancer and perineural invasion were independent predictors of inferior OS only in the SEER dataset, while completeness of cytoreduction (CC) and peritoneal carcinomatosis index were independent predictors for OS and PFS only in the Chinese center. Median OS was 24 months in CRC patients with PM alone and 36 months in those with both PM and OM (p = 0.181). Further, median PSF in patients with PM alone was 10 months, while that in individuals with both PM and OM was 20 months (p = 0.181). Conclusion: Surgical treatment of the primary and metastatic sites is effective and safe for CRC patients with PM. CC-0 is recommended for improved prognosis. Moreover, OM should be recognized as a feature of PM, and cytoreductive surgery combined with hyperthermic intraperitoneal chemotherapy is beneficial for CRC patients with OM.

Colorectal cancer (CRC) is one of the most commonly diagnosed cancers with an estimated 1.8 million new cases worldwide and associated with high mortality rates of 881 000 CRC-related deaths in 2018. Screening programs and new therapies have only marginally improved the survival of CRC patients. Immune-related genes (IRGs) have attracted attention in recent years as therapeutic targets. The aim of this study was to identify an immune-related prognostic signature for CRC. To this end, we combined gene expression and clinical data from the CRC data sets of The Cancer Genome Atlas (TCGA) into an integrated immune landscape profile. We identified a total of 476 IRGs that were differentially expressed in CRC vs normal tissues, of which 18 were survival related according to univariate Cox analysis. Stepwise multivariate Cox proportional hazards analysis established an immune-related prognostic signature consisting ofSLC10A2,FGF2,CCL28,NDRG1,ESM1,UCN,UTS2andTRDC. The predictive ability of this signature for 3- and 5-year overall survival was determined using receiver operating characteristics (ROC), and the respective areas under the curve (AUC) were 79.2% and 76.6%. The signature showed moderate predictive accuracy in the validation and GSE38832 data sets as well. Furthermore, the 8-IRG signature correlated significantly with tumour stage, invasion, lymph node metastasis and distant metastasis by univariate Cox analysis, and was established an independent prognostic factor by multivariate Cox regression analysis for CRC. Gene set enrichment analysis (GSEA) revealed a relationship between the IRG prognostic signature and various biological pathways. Focal adhesions and ECM-receptor interactions were positively correlated with the risk scores, while cytosolic DNA sensing and metabolism-related pathways were negatively correlated. Finally, the bioinformatics results were validated by real-time RT-qPCR. In conclusion, we identified and validated a novel, immune-related prognostic signature for patients with CRC, and this signature reflects the dysregulated tumour immune microenvironment and has a potential for better CRC patient management.

Gastrointestinal stromal tumors (GISTs), the most widespread type of sarcoma, contain driver gene mutations predominantly of receptor tyrosine kinase and platelet-derived growth factor receptor alpha. However, the inevitable development of resistance to imatinib (IM) cannot be fully attributed to secondary driver gene mutations. In this study, we investigated the role of microRNA-30a in sensitization of GIST cells to IM in vivo and in vitro. Higher levels of miR-30a were detected in GIST-T1 cells, which were more sensitive to IM than GIST-882 cells. IM treatment also reduced miR-30a levels, indicating the possible role of miR-30a in GIST IM resistance. Subsequently, miR-30a was confirmed to be an IM sensitizer via a mechanism that was attributed to its involvement in the regulation of cell autophagy. The interaction of miR-30a and autophagy in IM treated GIST cells was found to be linked by beclin-1. Beclin-1 knockdown increased IM sensitivity in GIST cell lines. Finally, miR-30a was confirmed to enhance IM sensitivity of GIST cells in mouse tumor models. Our study provides evidence for the possible role of miR-30a in the emergence of secondary IM resistance in GIST patients, indicating a promising target for overcoming this chemoresistance.