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Allograft rejection score strongly reflected anti-cancer immunity in breast cancer. a Boxplots of the allograft rejection score by single cells of the tumor, stromal cells, T cells, B cells, and myeloid cells in the GSE75688 cohort. p-values were calculated by the Kruskal-Wallis test. Boxplots were of Tukey type, with boxes depicting median and inter-quartile range. b Scatter plots between Allograft rejection score and CYT score in both cohorts. Spearman rank correlation was used for the analysis. c Correlation plots between score level of CYT and 8 immunerelated gene sets, including allograft rejection, coagulation, complement, interferon (IFN)-α, IFN-γ, IL6/JAK/STAT3/Signaling, and inflammatory response in the METABRIC and GSE96058 cohorts. The correlation value is indicated by color (blue for positive correlation and red for negative correlation), while the magnitude of the correlation is shown with circles.
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Although the value of tumor-infiltrating lymphocytes is well known, the clinical relevance of an increased immune response, specifically in breast cancer, has not been investigated across large cohorts of patients using computational algorithms. Our hypothesis stated that an enhanced immune response is associated with an improvement in outcomes. To...
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Context 1
... score in the tumor microenvironment (TME), the score was measured in a single-cell sequence cohort (GSE75688) that has transcriptomes of tumor cells, stromal cells, T cells, B cells, and myeloid cells. A strong separation in the score was seen between the immune cells. T cells and myeloid cells had higher scores than tumor and stromal cells ( Fig. 1a; p < 0.001). Next, we investigated how well the score reflected the immune response in the TME of breast cancer. We found that the score was strongly correlated with the cytolytic activity score (CYT), which reflects immune cell killing (Fig. 1b, Spearman rank test (r) = 0.892 and 0.860, respectively, both p < 0.01). The score was also ...
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... between the immune cells. T cells and myeloid cells had higher scores than tumor and stromal cells ( Fig. 1a; p < 0.001). Next, we investigated how well the score reflected the immune response in the TME of breast cancer. We found that the score was strongly correlated with the cytolytic activity score (CYT), which reflects immune cell killing (Fig. 1b, Spearman rank test (r) = 0.892 and 0.860, respectively, both p < 0.01). The score was also correlated with the other immune-related gene sets scores consistently in both METABRIC and GSE96058 cohorts, including complement, interferon (IFN)-γ response, IFN-α response, IL6/JAK/STAT3 signaling, and inflammatory response, but not with ...
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... 1b, Spearman rank test (r) = 0.892 and 0.860, respectively, both p < 0.01). The score was also correlated with the other immune-related gene sets scores consistently in both METABRIC and GSE96058 cohorts, including complement, interferon (IFN)-γ response, IFN-α response, IL6/JAK/STAT3 signaling, and inflammatory response, but not with coagulation (Fig. 1c). These findings suggest that the allograft rejection score strongly reflects anticancer immunity in ...
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... allograft rejection score reflected the amount of infiltrating immune cells, specifically anti-cancer immune cells Given that the allograft rejection score offers the strongest reflection of the anti-cancer immunity in the TME amongst all the Hallmark immune-related gene sets ( Fig. 1), we decided to focus on the allograft rejection score in breast cancer. In addition to immune function, it was of interest to investigate whether the allograft rejection score is associated with immune cell infiltration in the TME. The xCell algorithm was used to estimate immune cell infiltration, as performed in previous publications ...
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... we investigated the underlying mechanism involved in the score by using Gene Set Enrichment Analysis (GSEA) of ERpositive/HER2-negative breast cancer and TNBC. As expected, high score tumors enriched immune-related gene sets: inflammatory response, complement, interferon (IFN)-γ response, IFN-α response, tumor necrosis factor (TNF)-α signaling via NFkB, IL6/ JAK/STAT3 signaling, coagulation, p53 pathway, reactive oxygen species (ROS) pathway, and apoptosis in both subtypes in the METABRIC cohort ( Fig. 5b and Supplementary Fig. 1). In addition, we found that they also enriched pro-cancer gene sets: KRAS signaling up, PI3K/AKT/MTOR signaling, and apical surface gene sets. ...
Similar publications
Mingyi Yang Yani Su Ke Xu- [...]
Peng Xu
Objective
To investigate the ferroptosis-related long non-coding RNAs (FRLncs) implicated in influencing the prognostic and immune microenvironment in osteosarcoma (OS), and to establish a foundational framework for informing clinical decision making pertaining to OS management.
Methods
Transcriptome data and clinical data pertaining to 86 cases o...
Citations
... 16,17 Thus, targeted modulation of mitochondrial metabolism appears to be a potential tumor therapy. 18,19 Meanwhile, immune factors play a major role in breast cancer development, 20 and NDUFAF6 can promote PD-L1 expression by inhibiting the NRF2 signaling pathway to respond to the immune response. 21 In this research, we evaluated the levels of the mitochondria-related gene NDUFAF6 in breast cancer tissues and its association with prognosis. ...
Background
While NDUFAF6 is implicated in breast cancer, its specific role remains unclear.
Methods
The expression levels and prognostic significance of NDUFAF6 in breast cancer were assessed using The Cancer Genome Atlas, Gene Expression Omnibus, Kaplan-Meier plotter and cBio-Portal databases. We knocked down NDUFAF6 in breast cancer cells using small interfering RNA and investigated its effects on cell proliferation and migration ability. We performed gene expression analysis and validated key findings using protein analysis. We also assessed mitochondrial activity and cellular metabolism.
Results
NDUFAF6 was highly expressed in breast cancer, which was associated with a poorer prognosis. Knockdown of NDUFAF6 reduced the proliferation and migration ability of breast cancer cells. Transcriptome analysis revealed 2,101 differentially expressed genes enriched in apoptosis and mitochondrial signaling pathways. Western blot results showed NDUFAF6 knockdown enhanced apoptosis. In addition, differential gene enrichment analysis was related to mitochondrial signaling pathways, and western blot results verified that mitophagy was enhanced in NDUFAF6 knockdown breast cancer cells. JC-1 assay also showed that mitochondrial dysfunction and reactive oxygen species content were increased after knocking down NDUFAF6. In addition, basal and maximal mitochondrial oxygen consumption decreased, and intracellular glycogen content increased.
Conclusions
Knockdown of NDUFAF6 resulted in apoptosis and mitophagy in breast cancer cells and NDUFAF6 may be a potential molecular target for breast cancer therapy.
... Liver metastasis samples with a desmoplastic HGP were mainly characterized by an enrichment, relative compared to replacement HGP samples, of gene sets related to biological processes of inflammation and immune response (in decreasing order of relative enrichment): rejection of allografts, interferon gamma response, inflammatory response, interferon alpha response, IL2 STAT5 signaling, complement cascade, TNFA signaling via NFϰB, and IL6 STAT3 signaling during acute phase response. The hallmark gene set that best distinguished desmoplastic from replacement HGP samples, 'rejection of allografts', has been shown to accurately reflect the amount of tumor-infiltrating immune cells, cytolytic activity and to correlate with the expression of all other immune related hallmark gene sets [30]. Our deconvolution analysis (Xcell) also demonstrated a higher immune cell content in samples with a desmoplastic HGP compared to replacement HGP samples. ...
... Our deconvolution analysis (Xcell) also demonstrated a higher immune cell content in samples with a desmoplastic HGP compared to replacement HGP samples. In line with this, we have repeatedly, by immunohistochemical analyses, associated the desmoplastic HGP of liver metastases with the presence of an inflammatory cell infiltrate at the interface between liver tissue and the capsule that surrounds this type of metastases, while replacement-type liver metastases more frequently are immune deserts [10,30,31]. Interestingly, one of the genes that contributed considerably to the enrichment of inflammation and immune related hallmark gene sets in the desmoplastic HGP samples was GZMA, coding for granzyme A. Granzymes are serine proteases used by cytotoxic immune cells to exert cell killing. ...
The behaviour of metastases in patients with liver-metastatic colorectal cancer (CRC) is still not adequately considered during treatment planning. However, studies in large cohorts have shown that the disease course in these patients depends on the histopathological growth pattern (HGP) of the liver metastases, with the desmoplastic (or encapsulated) pattern responsible for a favourable outcome and the replacement pattern for an unfavourable course. To increase our knowledge of cancer biology in general as well as to design clinical trials that take into account the diverse behaviour of liver metastases, it is necessary to know the cellular and molecular determinants of these growth patterns. For that purpose, we compared the transcriptome of tumour tissue (prospective cohort; n = 57) sampled very precisely at the transition of metastasis and adjacent liver, between the desmoplastic and replacement HGP. In addition, the mutational profiles for 46 genes related to CRC were extracted from the RNA sequencing reads. First, we show that the genetic constitution of a liver metastasis from colorectal cancer does not determine its HGP. Second, we show clear differences between HGPs regarding the expression of genes belonging to the Molecular Signatures Database hallmark gene sets. Biological themes of the replacement HGP reflect cancer cell proliferation and glucose metabolism, while the desmoplastic HGP is characterized by inflammation and immune response, and angiogenesis. This study supports the view that HGPs are a reflection of the biology of CRC liver metastases and suggests the HGPs are driven epigenetically rather than by specific gene mutations.
... Furthermore, we have previously reported that several signaling pathways other than cell proliferation-related signaling were predominantly enhanced in AYA, including mTORC1, unfolded protein response, and PI3K/AKT/mTOR signaling using the pathway scoring method. [25][26][27] These findings were consistently observed in the two independent large cohorts. Interestingly, no significant differences in these signaling levels were observed among the other three age groups. ...
Background
We aimed to clarify the features of adolescents and young adults (AYA: younger than 40 years old) breast cancer (BC) compared with other age groups in estrogen receptor (ER)-positive/human epidermal growth factor receptor 2 (HER2)-negative BC, given the effects of age-related hormonal status.
Methods
The cohorts analyzed were divided into AYA (15-39 years old), perimenopausal (40-54 years old), menopausal (55-64 years old), and old (65+ years old). Clinicopathological and biological features were analyzed using gene set variation analysis and xCell algorithm using transcriptome profiles from large public databases of ER-positive/HER2-negative BC (METABRIC; n = 1353, SCAN-B; n = 2381).
Results
In the ER-positive/HER2-negative subtype, pathological lymph node positivity, and Nottingham grade 3 were higher among AYA (all P < 0.001). AYA patients had a trend toward worse disease-specific and overall survival, particularly compared with the perimenopausal group. Estrogen response late signaling decreased with age (all P ≤ 0.001 in both METABRIC and SCAN-B cohorts). AYA was associated with significantly higher BRCAness and DNA repair than the other groups (all P < 0.05 in both cohorts). AYA significantly enriched cell proliferation-related and procancerous gene sets [mTORC1, unfolded protein response, and phosphoinositide 3-kinase/protein kinase B/mammalian target of rapamycin (PI3K/AKT/mTOR) signaling] when compared with the others (all P < 0.03 in both cohorts). Interestingly, these features have also been observed in tumors <2 cm. Infiltration of CD8⁺, regulatory, T helper type 2 cells, and M1 macrophages was higher, while M2 macrophages were lower in AYA (all P < 0.03 in both cohorts). Finally, ER-positive/HER2-negative BC in AYA patients has different features of gene mutations, including AHNAK2, GATA3, HERC2, and TG, which were observed at a higher rate in AYA, and KMT2C, which was observed at a lower rate in AYA, compared with other age groups.
Conclusions
ER-positive/HER2-negative BC in AYA was highly proliferative with high immune cell infiltration compared with the other age groups.
... However, treatment outcome is still far from satisfactory. Certain subtypes of BC are likely to develop resistance to chemotherapy, leading to very poor therapeutic outcomes presenting a major hurdle for clinicians aiming to maximize patient survival [8]. And due to its complex and varied biological characteristics, none of the therapies have been proved to be beneficial [9]. ...
The escalating incidence of breast cancer (BC) in women underscores its grave health threat. Current molecular insights into BC's post-adjuvant therapy cure remain elusive, necessitating active treatment explorations. Immunotherapy, notably chemotherapy-induced immunogenic cell death (ICD), has emerged as a promising BC therapy. ICD harnesses chemotherapeutics to activate anti-tumor immunity via DAMPs, fostering long-term T-cell memory and primary BC cure. Besides chemotherapy drugs, Nanodrugs, traditional Chinese medicine (TCM) and ICIs also induce ICD, boosting immune response. ICIs, like PD-1/PD-L1 inhibitors, revolutionize cancer treatment but face limited success in cold tumors. Thus, ICD induction combined with ICIs is studied extensively for BC immunotherapy. This article reviews the mechanism of ICD related drugs in BC and provides reference for the research and development of BC treatment, in order to explore more effective clinical treatment of BC, we hope to explore more ICD inducers and make ICIs more effective vaccines.
Graphical abstract
... Liver metastasis samples with a desmoplastic HGP were mainly characterized by an enrichment, relative compared to replacement HGP samples, of gene sets related to biological processes of in ammation and immune response (in decreasing order of relative enrichment): rejection of allografts, interferon gamma response, in ammatory response, interferon alpha response, IL2 STAT5 signaling, complement cascade, TNFA signaling via NF B, and IL6 STAT3 signaling during acute phase response. The hallmark gene set that best distinguished desmoplastic from replacement HGP samples, 'rejection of allografts', has been shown to accurately re ect the amount of tumor-in ltrating immune cells, cytolytic activity and to correlate with the expression of all other immune related hallmark gene sets [29]. Our deconvolution analysis (Xcell) also demonstrated a higher immune cell content in samples with a desmoplastic HGP compared to replacement HGP samples. ...
... Our deconvolution analysis (Xcell) also demonstrated a higher immune cell content in samples with a desmoplastic HGP compared to replacement HGP samples. In line with this, we have repeatedly, by immunohistochemical analyses, associated the desmoplastic HGP of liver metastases with the presence of an in ammatory cell in ltrate at the interface between liver tissue and the capsule that surrounds this type of metastases, while replacement-type liver metastases more frequently are immune deserts [29][30][31]. Interestingly, one of the genes that contributed considerably to the enrichment of in ammation and immune related hallmark gene sets in the desmoplastic HGP samples was GZMA, coding for granzyme A. Granzymes are serine proteases used by cytotoxic immune cells to exert cell killing. ...
The behaviour of metastases in patients with liver-metastatic colorectal cancer (CRC) is still not adequately considered during treatment planning. However, studies in large cohorts have shown that the disease course in these patients depends on the histopathological growth pattern (HGP) of the liver metastases, with the desmoplastic (or encapsulated) pattern responsible for a favourable outcome and the replacement pattern for an unfavourable course. To increase our knowledge of cancer biology in general as well as to design clinical trials that take into account the diverse behaviour of liver metastases, it is necessary to know the cellular and molecular determinants of these growth patterns. For that purpose, we compared the transcriptome of tumour tissue (n = 57) sampled very precisely at the transition of metastasis and adjacent liver, between the desmoplastic and replacement HGP. In addition, the mutational profiles for 46 genes related to CRC were extracted from the RNA sequencing reads. First, we show that the genetic constitution of a liver metastasis from colorectal cancer does not determine its HGP. Second, we show clear differences between HGPs regarding the expression of genes belonging to the Molecular Signatures Database hallmark gene sets. Biological themes of the replacement HGP reflect cancer cell proliferation and glucose metabolism, while the desmoplastic HGP is characterized by inflammation and immune response, and angiogenesis. This study supports the view that HGPs are a reflection of the biology of CRC liver metastases and suggests the HGPs are driven epigenetically rather than by specific gene mutations.
... The MEXPRESS database was introduced to illustrate the association between ELF4 expression and its DNA methylation sites [19]. The SNV neoantigens, number of segments, fraction altered, aneuploidy score silent mutation rate, non-silent mutation rate, and fraction altered were obtained from the previous research [20]. ...
... For each step in CIC, ssGSEA was applied to exhibit differences among distinct groups. The xCell approach was utilized to compute the proportions of 64 distinct immune cell types as well as the scores of immunological, stromal, and tumor microenvironment (TME) components [20]. Aneuploidy score, silent mutation rate, non-silent mutation rate, fraction altered, number of segments, and SNV neoantigens were obtained from the previous research in the TCGA cohort [25]. ...
ELF4 (E74-like factor 4) is a transcription factor, dysregulation of which has been associated with carcinogenesis and cancer development. Nevertheless, the precise role of ELF4 in glioma pathology and its impact on clinical outcomes remains to be investigated. In the present research, comprehensive analyses demonstrated that elevated expression of ELF4 in glioma tissues correlates with malignant phenotypes and adverse clinical outcomes. Multivariate Cox regression analysis determined that ELF4 expression could serve as a reliable predictor of glioma outcomes. (CGGA, hazard ratio [HR]: 1.21, 95% confidence interval [CI]: 1.09-1.34, p<0.001; TCGA, HR: 1.19, 95%CI: 1.01-1.41, p=0.043; and Gravendeel, HR: 1.44, 95%CI: 1.15-1.80, p=0.002). Knockdown of ELF4 reduced the cell viability and migration capacity of glioma cells in vitro. In addition to the tumor invasive role, enrichment analysis revealed the overexpressed ELF4 was involved in the immune regulation, characterized by the elevated activity of Il6/Jak/Stat3 signaling, interferon alpha (IFN-α) response, and IL2/Stat5 signaling. Single-cell RNA sequencing (scRNA)-seq and spatial transcriptome (ST)-seq analyses revealed that ELF4 could induce reprogramming of tumor-associated monocytes/macrophages (TAMMs). Molecular docking analysis revealed ELF4 might be targeted by drugs/compounds, including Veliparib (ABT-888), Motesanib (AMG 706), and EHT 1864. Genomic analysis revealed that, in LGG, in the low ELF4 expression subgroup, IDH1 demonstrated a higher mutation rate, and TP53 and ATRX Chromatin Remodeler (ATRX) displayed the lower mutation rates, than the high ELF4 expression group.
Conclusion: Our research suggests that ELF4 may contribute to the prognostic assessment of glioma and personalized medicine.
... CYT is closely related with patient prognosis and outcomes and the expressions of both GZMA and PRF1 genes synergistically, or alone can affect the OS of cancer patients [24,[100][101][102] (Table 1). Recently, in breast cancer, it was reported that the overexpression of GZMA alone increased the infiltration of dendritic cells (DCs) and CD8+ T cells and was correlated with better OS, DSS, and progress-free interval (PFI) [104]. ...
Intratumoral immune cytolytic activity (CYT), calculated as the geometric mean of granzyme-A (GZMA) and perforin-1 (PRF1) expression, has emerged as a critical factor in cancer immunotherapy, with significant implications for patient prognosis and treatment outcomes. Immune checkpoint pathways, the composition of the tumor microenvironment (TME), antigen presentation, and metabolic pathways regulate CYT. Here, we describe the various methods with which we can assess CYT. The detection and analysis of tumor-infiltrating lymphocytes (TILs) using flow cytometry or immunohistochemistry provide important information about immune cell populations within the TME. Gene expression profiling and spatial analysis techniques, such as multiplex immunofluorescence and imaging mass cytometry allow the study of CYT in the context of the TME. We discuss the significant clinical implications that CYT has, as its increased levels are associated with positive clinical outcomes and a favorable prognosis. Moreover, CYT can be used as a prognostic biomarker and aid in patient stratification. Altering CYT through the different methods targeting it, offers promising paths for improving treatment responses. Overall, understanding and modulating CYT is critical for improving cancer immunotherapy. Research into CYT and the factors that influence it has the potential to transform cancer treatment and improve patient outcomes.
... The xCell algorithm obtained through the xCell website [48], was used to calculate the immune cell infiltration in the tumor microenvironment through transcriptomic data as we described previously [43,[49][50][51]. The scoring of homologous recombination, intratumor heterogeneity, fraction altered, silent mutation, non-silent mutation, single-nucleotide variant (SNV) neoantigens, indel neoantigens, leukocyte fraction, lymphocyte infiltration, and interferon (IFN)-response score was performed as published by Thorsson et al [52]. ...
Background
Hepatocellular carcinoma (HCC) with high Ki67 protein expression, the most commonly used cell proliferation marker, is associated with an aggressive biologic phenotype; however, conventional immunostaining is hampered by variability in institutional protocol, specific antibody probe, and by assessor subjectivity. To this end, we hypothesized that Ki67 gene (MKi67) expression would identify highly proliferative HCC, and clarify its association with oncologic outcome, tumor progression, and immune cell population in the tumor microenvironment (TME). Furthermore, we sought to identify the cell-cycle gene expression profile that confers this aggressive phenotype.
Methods
A total of 473 HCC patients with clinicopathological data associated with transcriptome were selected for this study: 358 patients from The Cancer Genome Atlas (TCGA) as the testing cohort, and 115 from GSE76427 as the validation cohort. Each cohort was divided into a highly proliferative group (MKi67-high) and the low MKi67 group (MKi67-low) by the median of Ki67 gene (MKi67) expression levels.
Results
MKi67-high HCC patients had worse disease-free survival (DFS), disease-specific survival (DSS), and overall survival (OS) independent of histological grade in the TCGA cohort. MKi67 expression correlated with histological grade and tumor size. MKi67 expression increased throughout the HCC carcinomatous sequence from normal liver, cirrhotic liver, early HCC, and advanced HCC. MKi67-high HCC was associated with higher intratumor heterogeneity, homologous recombination deficiency, and altered fraction as well as intratumoral infiltration of T helper type 1 (Th1) and Th2 cells, but lower interferon-gamma response and M2 macrophage infiltration. Cell proliferation-related gene sets in the Hallmark collection (E2F targets, G2M checkpoint, Myc target v1 and mitotic spindle), MTORC1 signaling, DNA repair, PI3K MTOR signaling, and unfolded protein response were all enriched in the MKi67-high HCC (false discovery rate (FDR) < 0.25).
Conclusions
High MKi67 gene expression identified highly proliferative HCC with aggressive biology involving classical pathways in cell cycle regulation and DNA repair, as well as poor overall oncologic outcomes. This suggests potential for personalized treatment strategies, but validation and refinement of these observations require further research to elucidate the underlying mechanisms and validate therapeutic targeting of these pathways in MKi67-high HCC tumors.
... BC is associated with risk variables such as family history, aging, and high breast density [6,7]. Certain subtypes of BC are likely to develop resistance to chemotherapy, leading to very poor therapeutic outcomes presenting a major hurdle for clinicians aiming to maximize patient survival [8]. Metastatic BC is typically inoperable, with treatments limited to chemotherapy, hormone therapy, targeted therapy, etc., with suboptimal efficacy [9,10]. ...
Background
Breast cancer is a major global health concern, and there is a continuous search for novel biomarkers to predict its prognosis. The mitochondrial protein NDUFAF6, previously studied in liver cancer, is now being investigated for its role in breast cancer. This study aims to explore the expression and functional significance of NDUFAF6 in breast cancer using various databases and experimental models.
Methods
We analyzed breast cancer samples from The Cancer Genome Atlas (TCGA), Gene Expression Omnibus (GEO), and Human Protein Atlas (HPA) databases, supplemented with immunohistochemistry (IHC) staining to assess NDUFAF6 expression. A breast cancer cell xenograft mouse model was used to evaluate tumor growth, apoptosis, and NDUFAF6 expression. Survival probabilities were estimated through Kaplan–Meier plots and Cox regression analysis. A Protein–Protein Interaction (PPI) network was constructed, and differentially expressed genes related to NDUFAF6 were analyzed using GO, KEGG, and GSEA. The relationship between NDUFAF6 expression, immune checkpoints, and immune infiltration was also evaluated.
Results
NDUFAF6 was found to be overexpressed in breast cancer patients and in the xenograft mouse model. Its expression correlated with worse clinical features and prognosis. NDUFAF6 expression was an independent predictor of breast cancer outcomes in both univariate and multivariate analyses. Functionally, NDUFAF6 is implicated in several immune-related pathways. Crucially, NDUFAF6 expression correlated with various immune infiltrating cells and checkpoints, particularly promoting PD-L1 expression by inhibiting the NRF2 signaling pathway.
Conclusion
The study establishes NDUFAF6 as a potential prognostic biomarker in breast cancer. Its mechanism of action, involving the inhibition of NRF2 to upregulate PD-L1, highlights its significance in the disease's progression and potential as a target for immunotherapy.
... Furthermore, genomic instability driven by mutations is a recognized catalyst for the initiation and aggressiveness of cancer cells. Utilizing the previously calculated scores by Thorsson and colleagues [28,35,36], our examination disclosed a noteworthy link between a myogenesis high GC and diminished silent and non-silent mutation frequencies, as well as copy number alteration (CNA) within the TCGA cohort (Supplementary Figure 2B). Nevertheless, there were no notable correlations with single nucleotide variant (SNV) neoantigens, intratumor heterogeneity, nor homologous recombination deficiency (HRD). ...
... Comprising proliferating cancer cells, stromal cells, vascular endothelial cells, infiltrating inflammatory cells, and various associated cells, the TME is a distinctive milieu that arises during tumor progression through interactions with the host [38]. We have previously reported that cells within the TME other than the cancer cells such as fibroblasts [39], immune cells [29,35], and adipocytes [27,40,41], and microvessel endothelial cells [33,42] are associated with cancer biology and prognosis. However, to date, myocytes has never been studied. ...