Purpose. Prognostic significance and gene signatures associated with carbonic anhydrase IX (CAIX) was investigated in triple negative breast cancer (TNBC) patients. Methods. Immunohistochemistry (IHC) for CAIX was performed in tissue microarrays (TMAs) of 136 TNBC patients. In a subset of 52 patients Digital Spatial Profiler (DSP) was performed in tumour (pancytokeratin+) and stroma (pan-cytokeratin-). Differentially expressed genes (DEGs) with P(+/- 0.25 and +/- 0.3, for tumour and stromal compartment, respectively) were identified. Four genes were validated at the protein level. Result. Cytoplasmic CAIX expression was independently associated with poor recurrence free survival in TNBC patients [hazard ratio (HR)=6.59, 95% confidence interval (CI): 1.47-29.58, P=0.014]. DEG analysis identified 4 up-regulated genes (CD68, HIF1A, pan-melanocyte, and VSIR) in the tumour region and 9 down-regulated genes in the stromal region (CD86, CD3E, MS4A1, BCL2, CCL5, NKG7, PTPRC, CD27, and FAS) when low versus high CAIX expression was explored. Employing IHC, high CD68 and HIF-1 alpha was associated with poorer prognosis and high BCL2 and CD3 was associated with good prognosis. Conclusions. DSP technology identified DEGs in TNBC. Selected genes validated by IHC showed involvement of CD3 and BCL2 expression within stroma and HIF-1 alpha, and CD68 expression within tumour. However, further functional analysis is warranted.

Colorectal cancer is the third most diagnosed malignancy worldwide and survival outcomes remain poor. Research is focused on the identification of novel prognostic and predictive biomarkers to improve clinical practice. There is robust evidence in the literature that inflammatory cytokine interleukin-6 (IL6) is elevated systemically in CRC patients and that this phenomenon is a predictor of poor survival outcome. However, evidence is more limited for the role of IL6 and its cognate receptor, IL6R, within the tumour epithelium and microenvironment. This study aimed to investigate IL6 and IL6R expression in a large cohort of retrospectively collected patient tumour specimens and determine association with clinical outcomes and characteristics. High expression of IL6R in the tumour epithelium was associated with reduced cancer-specific survival in patients with right-sided colon cancer. In these patients, high IL6R expression was also associated with an increased systemic neutrophil-to-lymphocyte ratio. A high number of copies of IL6 mRNA within the tumour-associated stroma, but not epithelium, was associated with reduced cancer-specific survival. The results from this study have validated IL6R as a marker of poor prognosis in a subgroup of CRC patients and identified the spatially resolved prognostic nature of intra-tumoural IL6 expression. This study has also highlighted the need for investigation of IL6/IL6R-targeted therapies as novel treatment strategies for patients with colon cancer.

ABSTRACT Purpose Carbonic anhydrase IX (CAIX) is a well‐established prognostic marker in breast cancer (BC). Nevertheless, this prognostic value is yet to be confirmed in BC subtypes. This study aims to investigate the prognostic effects of CAIX in oestrogen receptor (ER)‐negative (ER−) BCs and to establish pathways related to cytoplasmic CAIX expression in ER− and lymph node‐negative BCs. Methods Immunohistochemistry was performed to identify the prognostic role of CAIX protein expression in ER− tissue microarrays (TMAs) (n = 191). CAIX‐positive samples (n = 37) were transcriptionally profiled by TempO‐Seq and analysed by STRING. Real‐time quantitative PCR (RT‐qPCR) analysis was used to validate differentially expressed genes. Results Overexpression of cytoplasmic CAIX was an independent predictor of recurrence free survival, disease‐free survival and overall survival in ER− cohort. RNA transcriptomic analysis identified 10 significant genes in ER− cohort and 3 genes in the node‐negative group. The STRING database demonstrated a significant interaction between MUCL1 and GALNT6, which were linked with extracellular matrix organisation, degradation of the extracellular matrix and disease of glycosylation pathways. In the node‐negative group, SPNS2 is mainly involved in the sphingolipid de novo biosynthesis pathway. A significant correlation between cytoplasmic SphK1 and cytoplasmic hypoxia‐inducible factor‐1α was observed. Among the 10 genes, 7 genes (SERHL2, GALNT6, MUCL1, MMP7, PITX2, CEACAM6 and SPNS2) were selected, and their expression was quantitatively assessed by RT‐qPCR. The PCR data of these genes showed that SERHL2, GALNT6, MUCL1, PITX2, and SPNS2 mRNA levels were expressed in MDA‐MB‐231 BC cell lines at variable levels of hypoxic exposure. Conclusion Cytoplasmic CAIX was independently associated with poor prognosis in ER− BC. Gene expression profiles shed light on the pathways and genes associated with hypoxia in ER− BC. In node‐negative patients, SPNS2 was of particular interest.

Colorectal cancer (CRC) is a complex disease with key oncogenic pathways, including Wnt, MAPK, and PI3K, co-operating to drive tumour initiation and progression. Loss-of-function mutations in the Wnt-pathway inhibitor APC are the most prominent genetic alterations and are commonly seen as the tumour-initiating event. Here, we have used genetically engineered mouse models to introduce pathway-activating mutations of PI3K (Pik3ca, Pten) and MAPK (Kras) signalling to the mouse gut. Comprehensive characterization of these models reveals pathway-dependent cooperation, leading to marked allele dose-dependent acceleration of tumour formation, underpinned by MYC-driven transcriptional reprogramming and alterations in downstream signalling pathways. We find the amino acid transporter SLC7A5 to be highly upregulated upon activation of PI3K signalling. In human CRCs, SLC7A5expression correlates with the newly defined PDS1 pathway-derived subtype and highly proliferative tumours. Genetic depletion of Slc7a5 in the newly developed PI3K-hyperactive models drastically extends survival by delaying tumour formation, even in aggressive Kras/Pik3caco-mutant mice. Finally, Slc7a5 gene deletion sensitizes these models to targeted MAPK inhibition. Taken together, SLC7A5 drives progression of PI3K-mutant CRCs and is an attractive (co-)target for mutation-specific inhibitors.

Colorectal cancer (CRC) is a heterogenous malignancy underpinned by dysregulation of cellular signaling pathways. Previous literature has implicated aberrant JAK/STAT3 signal transduction in the development and progression of solid tumors. In this study we investigate the effectiveness of inhibiting JAK/STAT3 in diverse CRC models, establish in which contexts high pathway expression is prognostic and perform in depth analysis underlying phenotypes. In this study we investigated the use of JAK inhibitors for anti-cancer activity in CRC cell lines, mouse model organoids and patient-derived organoids. Immunohistochemical staining of the TransSCOT clinical trial cohort, and 2 independent large retrospective CRC patient cohorts was performed to assess the prognostic value of JAK/STAT3 expression. We performed mutational profiling, bulk RNASeq and NanoString GeoMx® spatial transcriptomics to unravel the underlying biology of aberrant signaling. Inhibition of signal transduction with JAK1/2 but not JAK2/3 inhibitors reduced cell viability in CRC cell lines, mouse, and patient derived organoids (PDOs). In PDOs, reduced Ki67 expression was observed post-treatment. A highly significant association between high JAK/STAT3 expression within tumor cells and reduced cancer-specific survival in patients with high stromal invasion (TSP high ) was identified across 3 independent CRC patient cohorts, including the TrasnSCOT clinical trial cohort. Patients with high phosphorylated STAT3 (pSTAT3) within the TSP high group had higher influx of CD66b + cells and higher tumoral expression of PDL1. Bulk RNAseq of full section tumors showed enrichment of NFκB signaling and hypoxia in these cases. Spatial deconvolution through GeoMx® demonstrated higher expression of checkpoint and hypoxia-associated genes in the tumor (pan-cytokeratin positive) regions, and reduced lymphocyte receptor signaling in the TME (pan-cytokeratin- and αSMA-) and αSMA (pan-cytokeratin- and αSMA +) areas. Non-classical fibroblast signatures were detected across αSMA + regions in cases with high pSTAT3. Therefore, in this study we have shown that inhibition of JAK/STAT3 represents a promising therapeutic strategy for patients with stromal-rich CRC tumors. High expression of JAK/STAT3 proteins within both tumor and stromal cells predicts poor outcomes in CRC, and aberrant signaling is associated with distinct spatially-dependant differential gene expression.

Background: Transforming growth factor β-activated protein kinase-1 (TAK1) plays an important role in MAPK and NFκB pathways and has been associated with colorectal cancer. The aim of this study was to determine how cytoplasmic and juxtanuclear punctate staining of TAK1 relates to immune checkpoint expression and cancer specific survival in colorectal cancer. Methods: Protein expression was assessed by immunohistochemistry on tissue microarrays from primary curative colorectal cancer resected specimens. Expression levels of cytoplasmic TAK1 by QuPath digital quantification and punctate TAK1 staining was scored using a manual point scoring technique and correlated with clinicopathological features, immune checkpoint expression and cancer-specific survival. Bulk RNA sequencing was performed in specimens to determine mutational profiles and differentially expressed genes. Results: A cohort of 875 patients who had undergone colorectal cancer resection were assessed for TAK1 expression. Higher levels of cytoplasmic TAK1 expression correlated with elevated PD1 and PD-L1 expression (p < 0.010). High punctate TAK1 expression was more commonly identified in poorly differentiated colorectal cancers (p = 0.036), had dysregulated mutational and transcriptional profiles with decreased insulin-like growth factor 2(IGF2) expression (p < 0.010), and independently predicted poor cancer-specific survival (HR 2.690, 95% CI 1.419–5.100, p = 0.002). The association of punctate TAK1 expression and recurrence remained after subgroup analysis for microsatellite-stable colorectal cancer (p = 0.028). Discussion: Punctate TAK1 expression is associated with worse cancer specific survival. TAK1 signalling may be an important pathway to investigate underlying mechanisms for recurrence in microsatellite-stable colorectal cancer.

AbstractColorectal cancer (CRC) is a heterogenous malignancy and research is focused on identifying novel ways to subtype patients. In this study, a novel classification system, tumour microenvironment score (TMS), was devised based on Klintrup–Mäkinen grade (KMG), tumour stroma percentage (TSP), and tumour budding. TMS was performed using a haematoxylin and eosin (H&E)‐stained section from retrospective CRC discovery and validation cohorts (n = 1,030, n = 787). TMS0 patients had high KMG, TMS1 were low for KMG, TSP, and budding, TMS2 were high for budding, or TSP and TMS3 were high for TSP and budding. Scores were assessed for association with survival and clinicopathological characteristics. Mutational landscaping and Templated Oligo‐Sequencing (TempO‐Seq) profiling were performed to establish differences in the underlying biology of TMS. TMS was independently prognostic in both cohorts (p < 0.001, p < 0.001), with TMS3 predictive of the shortest survival times. TMS3 was associated with adverse clinical features including sidedness, local and distant recurrence, higher T stage, higher N stage, and presence of margin involvement. Gene set enrichment analysis of TempO‐Seq data showed higher expression of genes associated with hallmarks of cancer pathways including epithelial to mesenchymal transition (p < 0.001), IL2 STAT5 signalling (p = 0.007), and angiogenesis (p = 0.017) in TMS3. Additionally, enrichment of immunosuppressive immune signatures was associated with TMS3 classification. In conclusion, TMS represents a novel and clinically relevant method for subtyping CRC patients from a single H&E‐stained tumour section.

Background: Tumor budding (TB), a single or cluster of up to 4 cells at the tumor-invasive front, is a well-established prognostic marker and an independent predictor of metastasis in colorectal cancer (CRC). Despite that, the underlying molecular mechanism that drives this phenotype and its relationship with the tumor microenvironment (TME) remains unclear. Methods: The relationship between TME and TB was investigated using multiplex immunofluorescence (mIF) in tissue microarrays (TMAs) of the tumor core and full sections. GeoMxTM digital spatial profiling (DSP) was employed to identify the mechanism underlying TB and its surrounding TME, using the GeoMx Immune-oncology pathway panel in full CRC sections. Results: In the current study, TB was independently associated with decreased cancer-specific survival (OR 1.856, 95% CI 1.360-2.533, P<0.001) in a cohort of 650 CRC patients. Additionally, TB was significantly associated with the Glasgow microenvironment score (GMS) (P=0.017). mIF performed in TMAs of the tumor core (n=650) showed that high CD3+ (OR0.374, 95% CI0.263-0.533, P<0.001) associated with good prognosis, whereas high CD68+ (OR2.902, 95% CI1.728-4.873, P<0.001) associated with poor prognosis. In addition, eighteen full sections were stained with an immune-related multiplex panel. Interestingly, in the invasive area, CD3+CD8+ was less frequent, while CD3+FOXP3+ was significantly higher compared to the stromal area in a patient with high TB (P=0.06, P<0.01, respectively). Moreover, though no significant difference was found, tumors with high buds showed an increase in neutrophil marker (CD66b) compared to low buds. No association is also found with a myeloid marker (CD68, CD163). The proliferation index of the tumor in the invasive area showed that the tumor with low buds is likely to be more proliferative. Besides, we found that a small subpopulation of budding is KI67+, suggesting the heterogeneity among TB. Furthermore, the mechanism underlying TB was observed using GeoMxTM. Twelve cases, selected from mIF stained, with low (n=6) or high buds (n=6) were utilized, and 96 areas of interest (AOI) were identified from the tumor core, invasive front, and stroma. The results showed the differential expression of genes between the tumor core and the invasive front in patients with high TB. A significant decrease in immune-related genes (e.g., CD3, NKG7, IL6, CXCR6, CD47, IFNAR1 and VSIR) compared to the stromal area at the invasive front was also observed (P<0.001). Conclusion: TB is associated with poor prognosis and showed a distinct gene expression characteristic compared to the tumor mass. It may also affect the inflammatory response at the invasive tumor. The underlying mechanism of TB and its relationship with TME warrants future studies to confirm this finding. Citation Format: Phimmada Hatthakarnkul, Holly Leslie, Hester Van Wyk, Leah Officer-Jones, Silvia Cusumano, Aula Ammar, Kathryn A.F. Pennel, Jean A. Quinn, Jennifer Hay, James Park, Noori Maka, John Le Quesne, Chanitra Thuwajit, Nigel Jamieson, Joanne Edwards. Understanding the molecular mechanism of tumor budding and its relationship with tumor microenvironment in colorectal cancer. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 5558.

220 Background: Microsatellite stable (MSS) colorectal cancer continues to have limited options for personalised therapeutic targets. The NFKB pathway is known to play an important role in inflammation-related carcinogenesis but has yet to be translated into therapies for the clinical patient. The aim of this study was to investigate the expression of cytoplasmic and punctate TAK1 (transforming growth factor β-activated protein kinase 1) in colorectal cancer and its relationship to immune checkpoint expression and prognosis. Methods: Patients undergoing primary colorectal cancer resection between 1997 and 2007 at Glasgow Royal Infirmary (UK) were studied for clinicopathological data and immunohistochemistry (IHC) performed on archival FFPE tissue from resected specimens. Antibodies for TAK1, PD-1, PD-L1, IKK alpha and other proteins were used for IHC, with digital analysis (QuPath) for quantification of cytoplasmic staining and punctate score for juxta-nuclear TAK1 assessment. Kaplan-Meier curves were created with log-rank test to determine survival. Cox-proportional hazards regression were used to determine multivariate hazard ratios (HR) and 95% confidence intervals (CI). Results: A total of 898 patients who underwent colorectal resection were identified. Higher TAK1 punctate expression was observed in left colon and rectal cancer, compared with right sided disease (p = 0.045). MMR proficient tumours had higher frequencies of high TAK1 punctate expression (p < 0.001). Both cytoplasmic and punctate TAK1 expression correlated with IKK expression (p < 0.050). High cytoplasmic TAK1 expression was associated with increased PD-1 and PD-L1 expression (p < 0.001). Punctate TAK1 expression was associated with worse survival (p = 0.037). These differences were accentuated in patients with MSS status (p = 0.016). On multivariate analysis, high punctate TAK1 expression remained a predictor of worse cancer-specific survival (HR 1.843, CI 1.129-2.956, p = 0.011). Conclusions: TAK1 expression was associated with MSI status, and higher TAK1 expression correlates with upregulated PD-1 and PD-L1 expression. High punctate TAK1 expression predicted cancer-specific survival. In subgroup analysis of MSS patients, high punctate TAK1 expression was associated with poor survival. Further interrogation into this pathway may identify inflammation-related therapeutic targets in MSS patients with colorectal cancer.

251 Background: Colorectal Cancer (CRC) remains a significant healthcare problem. Interest in the role of the Tumour Microenvironment (TME) in progression and metastasis is established, yet much remains to be learned about the mechanism of immune cell recruitment to the TME of primary and metastatic tumours. The Klintrup Makinen (KM) score measures the density of immune infiltrate to the invasive edge of tumours and is associated with good prognosis in primary CRC. Neutrophil infiltration to metastases is a poor prognostic feature suggesting phenotypic differences in the TME at primary and metastatic sites. We set out to examine epithelial determination of the TME in primary CRC. Methods: 180 patient’s primary CRC from a curated TMA underwent deep phenotyping using a multiplex-immunofluorescent panel (DAPI, Panck, CD3 (T cells), FOXP3 (T regulatory cells), CD68 (Macrophages), CD163 (Tumour associated macrophage), CD66b (Neutrophils), KI67). The Nanostring GeoMx Spatial Transcriptomic platform was used to obtain segmented (Tumour: Panck+; TME: Panck-) whole transcriptomic (18000 genes) readout from 54 matched cores from the same TMA. Results: No significant association was demonstrated between immune cell density of any cell types and clinicopathological variables including tumour side, MMR status, N-stage or survival. High KM grade was associated with high CD3 (P = 0.01) and FOXP3 (P = 0.002). Density of all immune cell types were highly corelated. Spatial transcriptomic analysis of the segmented epithelium of patients with high KM grade demonstrated over-expression of known neutrophil chemo-attractant CXCL5 (log2FC 3.76 Adj.P < 0.005). The TME compartments of cores with high CD68 and low CD163 counts were associated with adaptive immune co-ordination with high expression of CXCL13 (log2FC -2.97, Adj.P < 0.005) and CD74 (log2FC -1.42 Adj.P 0.007). IGF2 (log2FC 6.08 Adj.P < 0.005) and LCN15 (log2FC 4.96 Adj.P < 0.005) were globally over-expressed in both TME and epithelium of cores with generalised low immune infiltration. The epithelial compartment neutrophil dense cores also over-expressed IGF2 (log2FC -3.53 Adj.P 0.02). Conclusions: TMAs provide a unique opportunity for broad transcriptomic phenotyping of large numbers of patients simultaneously. The Nanostring GeoMx platform is a flexible tool which can be used in novel ways with other proteomic technologies to drive deeper analysis of the TME. CXCL5 expression in the epithelial compartment drives global immune infiltration to the TME and may be manipulated in different contexts therapeutically through CXCR2 signalling in future.

Background. Two pre-operative radiotherapy (RT) regimens are in common use for patients (pts) with locally advanced rectal cancer (LARC): Conventionally fractionated, long course chemoradiation (LCCRT, 25x1.8Gy with concomitant chemotherapy) or hypofractionated short-course RT (5X5Gy) followed by systemic chemotherapy (SCRT-C). RT is thought to induce immune responses to cellular damage, but the understanding of their evolution and relation to fractionation is limited. We are conducting a serial biospecimen collection in such pts with baseline, 2, 6 and 12 week (wk) sampling. We report serial measurement of tumor and peripheral blood lymphocyte responses for each regimen. Method. Multiplex immunofluorescence quantified tumor infiltrating lymphocytes (CD8, FOXP3). Routine diagnostic flow cytometry quantified circulating lymphocytes. A multiplex ELISA quantified cytokines in blood plasma. Bulk RNA-sequencing (QuantSeq) quantified gene expression within the tumor. Result. We report results for 20pts who received RT for stage III/IV LARC (13 LCCRT, 7 SCRT-C). Biopsy results are available for 10pts, circulating lymphocytes for 20pts and peripheral blood cytokines for 16pts. In LCCRT patients (n=8), relative to baseline, tumor infiltrating cytotoxic (CD8+) T cells were uniformly decreased during (2wks, P<0.05) and directly after treatment (6wks, P<0.05) before returning to baseline at 12wks. T regulatory cells (FOXP3+) similarly significantly decreased at 2wks and 6wks but remained below baseline at 12wks (P<0.05). Circulating lymphocytes also fell at wk2 and wk6 after commencing LCCRT and had begun recovering by wk12(n=13; P<0.05). The concentrations of circulating interleukins secreted by (IL2, IL8) or which activate T-lymphocytes (IL2, IL15) were reduced in the circulation at wk2 (n= 9; P<0.05) and wk6 (NS). In SCRT-C patients, we noted an increase in CD8 T cells at 2wks in 2/2 pts, also reflected in gene expression data, and an increase in FOXP3 T cells in 1/2 pts. Circulating lymphocytes were similarly decreased at wk2 in both the SCRT-C (n=7) and LCCRT (n=13) pts, but this reduction was less marked within the SCRT-C cohort at wk6 (P<0.01) and wk12 (P<0.05) relative to LCCRT patients. IL2, IL8 and IL15 did not change during or after SCRT-C (n=7; NS). Conclusion. LCCRT caused a drop in T cells during treatment, whilst SCRT-C appears to induce intra-tumoral T cell responses from wk2 and abrogates systemic reactions to a lesser extent. These results require evaluation in a larger cohort but have implications for understanding how RT induces microenvironmental changes and impacts pelvic bone marrow. We show in vivo that SCRT may be more immunostimulatory in LARC, with implications for trials combining RT with immunotherapy. Citation Format: Lily V. Hillson, Ross K. McMahon, Kathryn A. Pennel, Jean A. Quinn, Leia Jones, Raheleh Amirkhah, Aula Ammar, Phimmada Hatthakarnkul, Annabelle Ferguson, Simon W. Milling, Alec McDonald, Philip D. Dunne, Joanne Edwards, Sean M. O'Cathail, Campbell S. Roxburgh. Temporal changes in intratumoral and systemic lymphocytes in response to short and long course radiotherapy regimens in locally advanced rectal cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3231.

The genomic landscape of colorectal cancer (CRC) is shaped by inactivating mutations in tumour suppressors such as APC , and oncogenic mutations such as mutant KRAS . Here we used genetically engineered mouse models, and multimodal mass spectrometry-based metabolomics to study the impact of common genetic drivers of CRC on the metabolic landscape of the intestine. We show that untargeted metabolic profiling can be applied to stratify intestinal tissues according to underlying genetic alterations, and use mass spectrometry imaging to identify tumour, stromal and normal adjacent tissues. By identifying ions that drive variation between normal and transformed tissues, we found dysregulation of the methionine cycle to be a hallmark of APC-deficient CRC. Loss of Apc in the mouse intestine was found to be sufficient to drive expression of one of its enzymes, adenosylhomocysteinase (AHCY), which was also found to be transcriptionally upregulated in human CRC. Targeting of AHCY function impaired growth of APC-deficient organoids in vitro, and prevented the characteristic hyperproliferative/crypt progenitor phenotype driven by acute deletion of Apc in vivo, even in the context of mutant Kras . Finally, pharmacological inhibition of AHCY reduced intestinal tumour burden in Apc Min/+ mice indicating its potential as a metabolic drug target in CRC.

Purpose: Colorectal cancer (CRC) is the third most diagnosed cancer worldwide. Despite a well-established knowledge of tumour development, biomarkers to predict patient outcomes are still required. S100 calcium-binding protein A2 (S100A2) has been purposed as a potential marker in many types of cancer, however, the prognostic value of S100A2 in CRC is rarely reported.Material and Methods: In this study, immunohistochemistry (IHC) was performed to identify the prognostic role of S100A2 protein expression in the tumour core of the tissue microarrays (TMAs) in colorectal cancer patients (n=787). Bulk RNA transcriptomic data was used to identify significant genes compared between low and high cytoplasmic S100A2 groups. Multiplex immunofluorescence (mIF) was performed to further study and confirm the immune infiltration in tumours with low and high cytoplasmic S100A2. Results: Low cytoplasmic protein expression of S100A2 in the tumour core was associated with poor survival (HR 0.539, 95%CI 0.394-0.737, P<0.001) and other adverse tumour phenotypes. RNA transcriptomic analysis showed a gene significantly associated with the low cytoplasmic S100A2 group (AKT3, TAGLN, MYLK, FGD6 and ETFDH), which correlated with tumour development and progression. GSEA analysis identifies the enriched anti-tumour and immune activity group of genes in high cytoplasmic S100A2. Additionally, mIF staining showed that high CD3+FOXP3+ and CD163+ inversely associated with low cytoplasmic S100A2 (P<0.001, P=0.009 respectively).Conclusion: Our finding demonstrates a prognostic value of S100A2 together with the correlation with immune infiltration in CRC.

Triple‐negative breast cancer (TNBC) patients have the poorest clinical outcomes compared to other molecular subtypes of breast cancer. IL6/JAK/STAT3 signalling is upregulated in breast cancer; however, there is limited evidence for its role in TNBC. This study aimed to assess the expression of IL6/JAK/STAT3 in TNBC as a prognostic biomarker.

Colorectal cancer (CRC) is the third most common cancer worldwide. Poor survival of CRC associated with the development of tumour metastasis led to the investigation of the potential biomarkers to predict outcomes in CRC patients. Tumour budding (TB) is a well-known independent prognostic marker for poor survival and disease metastasis. Therefore, it has been suggested that TB status is included in routine clinicopathological factors for risk assessment in CRC. In contrast with a vast majority of studies regarding the prognostic power of TB, there is no clear evidence pertaining to the underlying molecular mechanism driving this phenotype, or an understanding of TB relationship with the tumour microenvironment (TME). The aim of the present study is to present a comprehensive review of TB and tumour cell signalling pathways together with the cross-talk of immune cells that could drive TB formation in CRC.

Nucleolin (NCL) is a multifunctional protein expressed in the nucleus, cytoplasm, and cell membrane. Overexpression of NCL has a controversial role as a poor prognostic marker in cancers. In this study, a meta-analysis was performed to evaluate the prognostic value of NCL in different subcellular localizations (cytoplasmic (CyNCL) and nuclear (NuNCL)) across a range of cancers. PubMed was searched for relevant publications. Data were extracted and analyzed from 12 studies involving 1221 patients with eight cancer types. The results revealed high total NCL was significantly associated with poor overall survival (OS) (HR = 2.85 (1.94, 4.91), p < 0.00001, I 2 = 59%) and short disease-free survival (DFS) (HR = 3.57 (2.76, 4.62), p < 0.00001, I 2 = 2%). High CyNCL was significantly associated with poor OS (HR = 4.32 (3.01, 6.19), p < 0.00001, I 2 = 0%) and short DFS (HR = 3.00 (2.17, 4.15), p < 0.00001, I 2 = 0%). In contrast, high NuNCL correlated with increased patient OS (HR = 0.42 (0.20, 0.86), p = 0.02, I 2 = 66%), with no significant correlation to DFS observed (HR = 0.46 (0.19, 1.14), p = 0.09, I 2 = 57%). This study supports the role of subcellular NCL as a poor prognostic cancer biomarker.

INTRODUCTION:Despite a well-known prognostic role in colorectal cancer, the genomic profiling of tumour budding remains to be elucidated. We aim to review the association of common mutations with tumour budding. METHODS:A systematic review of studies relating to tumour budding and genetic mutation in CRC was performed. The relationship between mutational status and tumour budding was evaluated using meta-analysis. RESULTS:A total of 6153 patients from 17 articles were included. According to the meta-analysis, high-grade tumour budding was significantly associated with KRAS mutation (OR = 1.52, 95 %CI: 1.13-2.02, P = 0.005) and MSS/pMMR (OR = 2.06, 95 %CI: 1.42-2.97, P = 0.0001). CONCLUSION:The significant association between high-grade tumour budding and mutated KRAS or MSS/pMMR may suggest a role of these mutations in the development of the tumour budding phenotype and be useful for stratifying patient outcome in CRC.