The National Cancer Research Institute (NCRI) is a UK-wide partnership between cancer research funders, which promotes collaboration in cancer research. Its member organizations work together to maximize the value and benefit of cancer research for the benefit of patients and the public. Rather than replace or duplicate any of the functions of its members, it seeks to add value through joint planning, coordination and collaboration..
Three-dimensional (3D) in vitro culture systems may better mimic in vivo physiological conditions, and are easily accessible methods to improve therapeutic effectiveness of human mesenchymal stem cells (MSCs), which with its many sources appear to harbor clinically relevant functional differences. We therefore investigated the impact and elucidated the mechanism(s) of 3D culture on the immunomodulatory capacity of two commonly used MSC sources, bone marrow (BM) and placental (P). In 3D conditions, PMSCs (PMSC 3D) form larger spheroids than BMMSCs (BMMSC 3D), with whole transcriptome profiling revealing significant enrichment of cell adhesion and immunomodulatory pathways. qPCR and functional validation demonstrated the highest expression of numerous key immunomodulatory factors and strongest capacity to inhibit T cell proliferation with PMSC 3D. Bioinformatics analyses predicted Intercellular Adhesion Molecule 1 (ICAM-1) as crucial for both PMSC 3D spheroid formation and enhanced immunomodulatory capacity, which was validated with flow cytometric analyses and further delineated with single-cell RNA sequencing data. To assess mechanistic involvement, we performed knockdown of ICAM-1 which significantly reduced PMSC 3D spheroid size as well as both in vitro and in vivo immunomodulatory capacity. These findings demonstrate that 3D culture significantly enhances the immunomodulatory potential of PMSCs, and reveal ICAM-1 as having a dual role in spheroid formation as well as modulation of immune responses. Our study also highlights the importance of understanding source-specific differences as well as the profound influence of 3D in vitro systems on MSC functions.
The tumor microenvironment (TME) not only influences cancer progression but also has been shown to have a significant effect on the prognosis. One of the major TME components is cancer-associated fibroblasts (CAFs), of which several subtypes have been identified in tumor tissues. Although single-cell RNA sequencing (scRNA-seq) technology is a powerful tool for investigating the proportions and composition of cells in tissues, large-scale datasets are needed to analyze small cell populations. Here, we constructed an integrated scRNA-seq dataset for non-small cell lung cancer (NSCLC) by compiling publicly available data and observed differences in TME heterogeneity and cellular composition between lung adenocarcinoma (LUAD) and lung squamous cell carcinoma (LUSC). In particular, we identified significant differences in CAF subtypes between LUAD and LUSC. Inflammatory CAFs (iCAFs) were predominantly found in LUAD, whereas myofibroblastic CAFs (mCAFs) were more common in LUSC. A coculture analysis of lung fibroblasts confirmed that LUAD cells induced the iCAF phenotype and, in contrast, that LUSC cells promoted myofibroblastic differentiation. A correlation analysis with the prognosis identified mCAFs as poor prognostic factors in both LUAD and LUSC, but iCAFs were found to be a poor prognostic factor only in LUSC, with the opposite pattern observed in LUAD. This work highlights important considerations regarding the CAF subtype dominance in LUAD and LUSC, which may be related to the prognosis.
Elucidation of the molecular mechanism underlying metastatic dissemination in patients with high-grade serous ovarian carcinoma (HG-SOC) has the potential to affect patient outcome. This study explores the role of gasdermins (GSDMs) in HG-SOC, focusing on novel pyroptosis-independent nuclear functions of GSDME, which are integrated with the endothelin-1 (ET-1)/ET-1 receptor A (ETAR) signaling to sustain metastatic progression. In this tumor, GSDME upregulation is correlated to epithelial-mesenchymal transition (EMT) and ETAR expression. ET-1 signaling fuels GSDME expression by inducing its transcription via the core EMT factors, ZEB1 and ZEB2. GSDME, in turn, translocates to the nucleus to engage ZEB1 and transcriptionally regulate genes coupled with EMT and inflammatory signals, such as E-cadherin, vimentin and interleukin (IL)-6. GSDME depletion, similarly to ZEB1 and ETAR blockade, restrains ET-1-induced EMT phenotypic plasticity and inflammatory cytokine release. Clinically relevant, ET-1 receptor (ET-1R) antagonist, by depleting the nuclear reservoir of the GSDME/ZEB1 transcriptional complex, hinders the metastatic traits of HG-SOC. The intertwined ETAR/GSDME/ZEB1 circuitry characterizes mesenchymal HG-SOC patients and associates with a high-risk of poor survival. Together, these findings unveil GSDME as a key transcriptional regulator of aggressive behaviors and worse prognosis in HG-SOC patients, in an ET-1-driven alliance with ZEB1, which could be targeted by ET-1R antagonist to reduce the metastatic burden of this tumor.
Medulloblastoma, the most common malignant brain tumor of childhood, exhibits significant biological complexity that demands deeper exploration. Here, we present a large multiomics dataset integrating data from 384 primary medulloblastoma patient samples across five omic layers: CpG methylome, transcriptome, proteome, phosphoproteome, and metabolome, paired with associated clinical metadata. Data integration revealed intertumoral heterogeneity of lipid metabolism across proteomic subtypes. Notably, while the MYC-FASN-SCD axis drives lipid biosynthesis, pathway inhibition elicits a compensatory escape mechanism in vivo through exogenous fatty acid uptake. Unexpectedly, we demonstrated that MYC triggers lipid storage, creating a unique dependency on lipid droplet-mitochondria communications to sustain tumor maintenance in vivo. Together, this comprehensive analysis reveals a targetable vulnerability downstream of MYC that constitutes a promising therapeutic approach to treat currently untreatable medulloblastoma subtypes.
Background Hepatocellular carcinoma has poor prognosis due to its high recurrence rate, even after curative surgery. Epigenetic regulators are known to play a critical role in cancer progression, and the histone methyltransferase SETD8/KMT5A has been reported to be overexpressed in various malignancies. In this study, we aimed to elucidate the role of SETD8/KMT5A in hepatocellular carcinoma. Methods We investigated SETD8/KMT5A expression in 345 primary hepatocellular carcinoma resection specimens through immunohistochemical staining. For functional analyses, we conducted a loss-of-function study of SETD8/KMT5A using hepatocellular carcinoma cell lines, including in vitro assays for proliferation, cell cycle, invasion, and RNA sequencing with gene ontology analysis. Additionally, we performed xenograft experiments in mice and performed similar experiments using the SETD8/KMT5A inhibitor UNC0379. Results All cases were divided into either the SETD8 high-expression (n = 197) or low-expression (n = 148) groups. The high-expression group exhibited significantly poorer 5-year overall survival and 2- and 5-year disease-free survival compared with the low-expression group (both p < 0.001). Multivariate analysis indicated that high SETD8 expression was an independent poor prognosis factor in overall (p = 0.0255) and disease-free (p = 0.0051) survival. SETD8/KMT5A knockdown suppressed proliferation by inhibition of G1 to S phase transition (p < 0.001). Gene ontology terms were related to cancer progression, including cell adhesion, MAPK-related signaling and chromatin remodeling. SETD8/KMT5A knockout using CRISPR/Cas9 inhibited tumor growth (p < 0.01) in vivo. Conclusions SETD8/KMT5A overexpression was associated with poor prognosis and was an independent prognostic factor in hepatocellular carcinoma. In vitro and in vivo analysis, the inhibition of SETD8 could repress hepatocellular carcinoma progression through the regulation of cell activity and cell cycle transition.
