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, I² = 59%) and short disease-free survival (DFS) (HR = 3.57 (2.76, 4.62), p < 0.00001, I² = 2%). High CyNCL was significantly associated with poor OS (HR = 4.32 (3.01, 6.19), p < 0.00001, I² = 0%) and short DFS (HR = 3.00 (2.17, 4.15), p < 0.00001, I² = 0%). In contrast, high NuNCL correlated with increased patient OS (HR = 0.42 (0.20, 0.86), p = 0.02, I² = 66%), with no significant correlation to DFS observed (HR = 0.46 (0.19, 1.14), p = 0.09, I² = 57%). This study supports the role of subcellular NCL as a poor prognostic cancer biomarker.

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.

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.

Purpose: Two dominant neoadjuvant radiotherapy (RT) regimens are in common use for locally advanced rectal cancer (LARC): long-course RT (LCRT; 25 × 1.8 Gy) with concomitant chemotherapy and short-course RT (SCRT; 5 × 5 Gy), typically followed by systemic chemotherapy. This study uses serial sampling to investigate the evolution of systemic and local immune responses to different RT regimens in LARC. Experimental Design: We conducted a serial sampling study involving patients receiving RT for LARC, in which longitudinal blood and tumor biospecimens were collected at baseline and at 2, 6, and 12 weeks after treatment initiation. Leukocyte concentrations via full blood count and a multiplex cytokine ELISA of plasma samples measured systemic changes. Multiplex immunofluorescence (CD8 and FOXP3) and RNA sequencing of tumor biopsies were used to assess local changes. Results: Circulating lymphocyte concentrations tended to increase in SCRT patients and decrease in LCRT patients between week 2 and week 6 and were significantly lower in LCRT patients compared with SCRT patients at week 6 (P < 0.0001) and week 12 (P = 0.019). Additionally, we report higher densities of CD8+ and FOXP3+ tumoral lymphocytes from SCRT patients compared with LCRT patients at week 2 (CD8 P = 0.053; FOXP3 P = 0.023) and week 6 (CD8 P = 0.035; FOXP3 P = 0.0016). Conclusions: SCRT is less lymphodepleting and induces more frequent increases in intratumoral T-cell infiltration compared with LCRT. These results are relevant to the field of radiation–immune-oncology combination studies in LARC. Furthermore, these findings may underpin early trial results, in which higher rates of response to RT–immune checkpoint inhibitor combinations have been reported with SCRT-based regimens.

Background: Tumour budding (TB), defined as a small cluster of up to four cells at the invasive front of the tumour, is a well-established independent and robust prognostic biomarker in colorectal cancer (CRC). This is strongly associated with adverse clinicopathological features and poor survival outcomes. Despite its clinical relevance, the precise underlying mechanism responsible for TB phenomenon remains unclear. Methods: Multi-omic approaches from bulk, regional GeoMx and Spatial Molecular Imager (SMI) RNA were used to identify the underlying mechanism of TB and its possible correlation with tumour microenvironment (TME) in CRC tissue. The results were validated using immunohistochemistry (IHC) and multiplex immunofluorescence (mIF) staining. Results: Patients with high TB experience worse outcomes and associate with adverse clinical factors across two independent CRC cohorts. Bulk and regional RNA expression analyses reveal that tumours with high TB are significantly enriched for TNF-α and TGF-β signatures in both cohorts. Single cell CosMx SMI analysis confirmed TB cells exhibit higher expression of these signatures than adjacent invasive edge tumour cells. Elevated cyclinD1 expression was also observed within TB, and high cyclinD1 levels tend to experience poorer CRC prognosis. Furthermore, regional bulk RNA expression within the non-tumour (PanCK-) invasive edge areas demonstrated that tumours exhibiting high TB revealed the significantly differential expressions of immune-related genes (e.g. CD3 , NKG7 , IL6 , CXCR6 , CD47 , IFNAR1 and VSIR ). Single cell CosMx SMI analysis revealed that cancer-associated fibroblasts (CAFs) were physically the closest cells to TB cells. This spatial proximity was confirmed at the protein level using mIF, where the distance from TB to CD68+ macrophages predicted significantly poorer CRC outcomes. Conclusion: This multi-omic study confirms the prognostic significance of TB in CRC patients across two independent cohorts. Our findings highlight that TNF-α and TGF-β signalling play a crucial role in budding cells development by regulating cyclinD1. Furthermore, the transcriptomic analysis reveals an immunosuppressive niche characterised by reduced immune activity and close spatial interactions with CAFs and macrophages Ultimately, this study provides valuable insight into TB’s underlying mechanism and its complex interactions within the TME. This could provide a foundation for developing targeted therapeutic strategies in CRC.