High Expression Group Research Articles

Correlation analysis of dynamic enhanced energy spectrum CT parameters with Ki-67 high expression in hepatocellular carcinoma

Objective: To investigate the correlation between dynamic enhanced energy spectral CT parameters and Ki-67 high expression in hepatocellular carcinoma (HCC). Methods: A cross-sectional study. This retrospective case-control study analyzed the clinical data of 101 patients with pathologically confirmed HCC in Xiamen Hospital of Traditional Chinese Medicine and Zhongshan Hospital of Xiamen University from December 2017 to March 2023. These 101 patients included 84 males and 17 females, and the age[M(Q1, Q3)] was 59.0(49.0,66.0)years. These patients were divided into two groups according to the immunohistochemical Ki-67 expression levels in tumor tissues: the high expression group (Ki-67%>20%, n=59) and the low expression group (Ki-67%≤20%, n=42).CT values on 70 keV and 140 keV monochromatic energy images (HU70 keV-a, HU140 keV-a, HU70 keV-p, HU140 keV-p) and water density (Dwater-a, Dwater-p) were measured in arterial phase and portal vein phase, and the difference of HU70 keV, HU140 keV, Dwater values between portal vein and arterial phase (ΔHU70 keV, ΔHU140 keV, ΔDwater), as well as ratio of HU70 keV, HU140 keV, Dwatervalues between portal vein and arterial phase (HU70 keVratio, HU140 keVratio, Dwaterratio) were calculated. Spearman correlation analysis was used to analyze the correlation between the CT spectral parameters and Ki-67%. Multivariate logistic regression model was used to determine the factors associated with high expression of Ki-67. The receiver operating characteristics (ROC) curves were used to indicate the efficacy of dynamic enhanced spectral CT in evaluating Ki-67 high expression in HCC. Results: The high Ki-67 expression group revealed higher alpha fetal protein levels, larger tumor diameter and more irregular tumor shape compared with the low Ki-67 expression group,and the differences were statistically significant (all P<0.05). Spearman correlation analysis showed that the HU140 keV-p, Dwater-p, HU70 keV ratio, HU140 keV ratio, Dwater ratio, ΔHU70 keV, ΔHU140 keV, ΔDwater were positively correlated with Ki-67 positivity rate (r:0.31-0.50, all P<0.05). The spectral CT parameters (HU70 keV-p, HU140 keV-p, Dwater-p, HU70 keV ratio, HU140 keV ratio, Dwater ratio, ΔHU70 keV, ΔHU140 keV, ΔDwater) in high Ki-67 expression group were significantly higher than those in low Ki-67 expression group (all P<0.05). Multivariate logistic regression model shows that Dwater-p(OR=1.16, 95%CI: 1.05-1.29, P=0.005), ΔHU140 keV(OR=1.39, 95%CI: 1.20-1.62, P<0.001) and irregular tumor morphology (OR=5.25, 95%CI: 1.61-17.12, P=0.006) were correlative factors for high Ki-67 expression. The HU140 keV ratio and ΔHU140 keV alone evaluated the highest AUC of high Ki-67 high expression in HCC, which were 0.82 (95%CI: 0.74-0.90), the sensitivity were 61.0%, and the specificity were 88.1% and 85.7%. The combined analysis of Dwater-p, ΔHU140 keV and irregular tumor morphology had an increased AUC of 0.88 (95%CI: 0.81-0.95) in assessment high Ki-67 expression, with the sensitivity of 84.7% and the specificity of 78.6%. Conclusions: Dynamic enhanced spectral CT parameters were positively correlated with the Ki-67 expression in HCC. Spectral CT provides a non-invasive method to evaluate the proliferation status of HCC cells, and the efficiency could be improved by multi-parameter analysis combining spectral CT parameters and morphologic features.

High expression of ARPC1B correlates with immune infiltration and poor outcomes in glioblastoma

ObjectiveTo investigate the role of ARPC1B in GBM and its prognostic value. MethodsmRNA and protein expression of ARPC1B in GBM was analyzed using the TCGA; TIMER2 and the HPA databases, and protein expression differences were detected using immunohistochemistry. K-M analysis and Cox regression analysis were performed on high and low ARPC1B expression groups in the TCGA database. The relationship between immune cells and ARPC1B expression was explored using the TIMER2 database. GO and KEGG analyses were conducted to investigate the functions of ARPC1B-related genes in GBM. ResultsARPC1B was highly expressed in both GBM tissues and cell lines, and it was demonstrated as a prognostic biomarker for GBM. ARPC1B expression levels showed associations with immune cell populations within the GBM microenvironment. ConclusionARPC1B can regulating immune infiltration in the GBM microenvironment, indicating its potential as a novel therapeutic target for GBM.

Correlation between IL3 signaling pathway-related genes and immune checkpoint inhibitor efficacy in patients with renal cell carcinoma.

There is a lack of effective biomarkers that predict immunotherapy efficacy in clear cell renal cell carcinoma(KIRC). We aimed to identify biomarkers that would predict the efficacy of KIRC treatment with immune checkpoint inhibitors (ICIs). Cohort data of KIRC patients with somatic mutations, mRNA expression and survival data from The Cancer Genome Atlas (TCGA) database and immunotherapy cohort and Genomics of Drug Sensitivity in Cancer (GDSC) database were analyzed and divided into interleukin 3 (IL3) pathway-related genes high expression (IL3-High) and IL3 pathway-related genes low expression (IL3-Low) groups according to pathway expression status to assess the relationship between the IL3 pathway-related genes activation status and the prognosis of KIRC patients treated with ICIs. The data were validated by immunohistochemistry experiments, and possible mechanisms of action were explored at the level of gene mutation landscape, immune microenvironment characteristics, transcriptome and copy number variation(CNV) characteristicsRESULTS: The IL3 pathway-related genes was an independent predictor of the efficacy of ICIs in KIRC patients, and the IL3-High group had a longer overall survival (OS); KIRC patients in the IL3-High group had increased levels of chemokines, cytolysis, immune checkpoint gene expression and abundant immunity. The IL3-Low group had poor immune cell infiltration and significant downregulation of complement activation, cytophagy, B-cell activation, and humoral immune response pathways. The high group was more sensitive to targeted drugs of some signaling pathways, and its efficacy in combining these drugs with immunity has been predicted in the published literature. The IL3 pathway-related genes can be used as a predictor of the efficacy of ICIs in KIRC. The IL3 pathway-related genes may affect the therapeutic efficacy of ICIs by affecting the expression of immune-related molecules, immune cell infiltration, and the level of immune response pathways.

An immune-related prognostic gene ULBP2 is correlated with immunosuppressive tumor microenvironment and immunotherapy in breast cancer

Breast cancer (BC) is one of the major dangerous tumors threatening women's lives. We here aimed to sort out prognostic immune-related genes by univariate Cox regression analysis and build a model of immune-related genes for forecasting the prognosis of BC patients. We identified UL16 binding protein 2 (ULBP2) as a valuable gene for study in the model using related databases and algorithms analysis. We found the stromal and immune cells scores were higher in ULBP2 high expression group and ULBP2 was related to kinds of immune cells, most importantly had negative correlation with CD8+ T cell. Notably, ULBP2 was positively correlated with tumor mutational burden (TMB) and had relationship with many immune checkpoints. Correlation analysis revealed that ULBP2 expression was closely linked to the clinicopathological characters and negatively related to BC patient survival. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis showed the functional enrichment of differential genes related to ULBP2. Gene Set Enrichment Analysis (GSEA) indicated pathway enrichment in ULBP2 high and low expression groups. In short, this study comprehensively investigated the potential function of ULBP2 in BC, which might make ULBP2 to be an important therapeutic target for BC.

DJ-1: A Potential Biomarker Related to Prognosis, Chemoresistance, and Expression of Microenvironmental Chemokine in HR-Positive Breast Cancer.

DJ-1 is significantly elevated in various malignancies. However, the clinical significance of DJ-1 in hormone receptor (HR)-positive (HR+) breast cancer remains unclear. We evaluated DJ-1 expression in different databases and validated in vitro assay by RT-PCR and western blot among HR+ breast cancer. The correlations between DJ-1 level and tumor-immune were calculated. Mutational landscape, enriched signaling pathways, and drug sensitivity analyses were also assessed between DJ-1 high and low-expression groups. DJ-1 was upregulated in HR+ breast cancer, and high DJ-1 expression was significantly linked with poor prognosis. DJ-1 was correlated with the expression and function of different immune cells. The low DJ-1 group showed sensitivity to paclitaxel and docetaxel, while the high-expression group showed sensitivity to doxorubicin. CTLA4 and PD-L1 were more sensitive in high-DJ-1 group. It is involved in a range of pathways and might behave as a novel biomarker of prognostic value for the immune environment and drug sensitivity in HR+ breast cancer.

Comprehensive analysis of m6A reader YTHDF2 prognosis, immune infiltration, and related regulatory networks in hepatocellular carcinoma

BackgroundN6-Methyladenosine (m6A) RNA modification is the most prevalent internal modification pattern in eukaryotic mRNAs and plays critical roles in diverse physiological and pathological processes. However, the expression of m6A regulator YTHDF2, its prognostic value, its biological function, its correlation with tumor microenvironment (TME) immune infiltrates, and related regulatory networks in hepatocellular carcinoma (HCC) remain determined. MethodsTCGA, GTEx, and GEO databases were used to investigate the expression profile of YTHDF2 in HCC. We performed differentially expressed genes (DEGs) analysis and constructed a PPI network to explore the biological processes of YTHDF2 in HCC. Kaplan-Meier curves and Cox regression analysis were used to assess the prognostic value of YTHDF2 and then a clinical prognostic nomogram was constructed. Additionally, ssGSEA was performed to assess the correlation between YTHDF2 and immune infiltration levels. The TISIDB database was applied to explore the expression of YTHDF2 in immune and molecular subtypes of HCC. GSEA identifies the YTHDF2-related signaling pathways. Finally, we utilized miRNet and starBase database to construct regulatory networks for HCC based on lncRNA-miRNA and miRNA-YTHDF2 interactions. ResultsYTHDF2 was significantly upregulated in HCC tumor tissues compared with the adjacent normal tissues. HCC patients in the high YTHDF2 expression group had poorer survival. Multivariate Cox analysis suggested that YTHDF2 may be a new independent prognostic indicator for HCC patients, with the prognostic nomogram exhibiting satisfactory results. YTHDF2 expression was significantly correlated with TME immune cell-infiltrating characteristics. Strong correlations were also shown in immune subtypes, molecular subtypes and immune checkpoints. Further analysis revealed that the combination of YTHDF2 expression and immune cell score was considerably associated with survival outcome in HCC patients. GESA analysis demonstrated that high YTHDF2 expression is associated with multiple biological processes and oncogenic pathways. Moreover, 14 possible regulatory networks were constructed, which are associated with HCC progression. ConclusionOur findings revealed that YTHDF2 may serve as a promising prognostic biomarker for HCC and may regulate the tumor immune microenvironment to provide effective therapeutic strategies.

Expression and Clinical Significance of Exosome Component 4 in Newly Diagnosed Patients with Diffuse Large B-Cell Lymphoma

To explore the expression of Exosome Component 4（EXOSC4） in the tissues of newly diagnosed patients with diffuse large B-cell lymphoma (DLBCL) and its clinical significance. The expression of EXOSC4 protein in the tissues of 181 newly diagnosed DLBCL patients was analyzed by immunohistochemical staining. Clinical data were collected. The correlation between EXOSC4 protein expression in the tissues of newly diagnosed DLBCL patients and clinical features were analyzed and its prognostic significance. The positive rate of EXOSC4 protein expression was 68.51% in the tissues of 181 newly diagnosed DLBCL patients. These patients were divided into two groups, with 44 cases in high expression group and 137 cases in low expression group. There were no significant differences in age, gender, B symptoms, serum lactate dehydrogenase (LDH) level, Eastern Cooperative Oncology Group (ECOG) score, Ann Arbor stage, extranodal disease, International Prognostic Index (IPI) score, National Comprehensive Cancer Network IPI (NCCN-IPI) score, and cell origin between the two groups (P>0.05). Cox multivariate regression analysis showed that high EXOSC4 protein expression in tissues was an independent poor prognostic factor for OS and PFS in newly diagnosed DLBCL patients (all P<0.05). K-M survival analysis showed that newly diagnosed DLBCL patients with high EXOSC4 protein expression had significantly shorter overall survival (OS) and progression free survival (PFS) than those patients with low EXOSC4 protein expression (all P<0.05). High EXOSC4 protein expression in tissues of newly diagnosed DLBCL patients is an independent poor prognostic factor for survival.

Identification of enhancer RNA AC005515.1 as a novel biomarker for prognosis in esophageal cancer and predictors of immunotherapy response.

The enhancer RNA (eRNA) signature shows excellent promise in the prognostic role of many malignancies, but its value has not been fully explored in esophageal cancer (ESCA). We comprehensively analyzed 33 oncogene expression matrices and clinical data from The Cancer Genome Atlas (TCGA) and identified ESCA prognostic-related key eRNAs by Kaplan-Meier and co-expression analysis. We also investigated the prognostic role of the key eRNA using a series of bioinformatics approaches, including immune infiltration, immune function, immune subtypes, and the tumor microenvironment. Finally, the tumor immune dysfunction and exclusion (TIDE) score was used to predict the immune response to immune checkpoint blockade (ICB) therapy. We identified eRNA AC005515.1, AC012368.1, AP003469.2, Clorf61, and WDFY3-AS2 were associated with the prognosis of ESCA. AC005515.1 is a critical prognostic-related eRNA in ESCA and was significantly co-expressed with immune checkpoint genes (CTLA4, CD274, etc.). In the pan-cancer analysis, AC005515.1 was also associated with the prognosis of seven cancers, including kidney renal papillary cell carcinoma and low-grade brain glioma. It was also found to be co-expressed with immune checkpoint genes in these tumors. Moreover, high expression of AC005515.1 was associated with CD8+ T cells and M1 macrophages infiltration, and the AC005515.1 high-expression group had a higher TIDE score in ESCA. Overall, eRNA AC005515.1 is associated with the local immune environment of ESCA and may become a new biomarker of ESCA prognosis and immunotherapy response.

Abstract A033: Secretogranin V as a Potential Biomarker for Esophageal Squamous Cell Carcinoma

Abstract Background: Oesophageal squamous cell carcinoma (ESCC) remains one of the most poorly diagnosed and deadly cancers worldwide. Identification of biomarkers to accurately predict the risk of recurrence and survival after surgery is therefore crucial to improving patient outcomes. This study examined the expression of Secretogranin V (SCG5) and its correlation with the prognosis of patients with ESCC. Methods: Transcription levels of SCG5 were evaluated in 22 cell lines of ESCC. The biological roles of SCG5 in cell invasion, proliferation, and migration were verified by small interfering RNA- mediated knockdown experiments. The expression of SCG5 was measured in 165 ESCC tissues by using quantitative reverse-transcription (qRT)-PCR, and its association with clinicopathological parameters was analyzed. Results: SCG5 mRNA expression levels varied widely in ESCC cell lines. Knockdown of SCG5 expression significantly suppressed cell invasion, proliferation, and migration of ESCC cells in vitro. Analysis of clinical specimens revealed that the expression of SCG5 mRNA was overexpressed in the ESCC compared to the adjacent normal oesophageal tissues. The high SCG5 expression group had significantly shorter overall and disease-free survival times. In the multivariable analysis, the high expression of SCG5 was determined to be an independent poor prognostic factor. Conclusion: SCG5 may have a significant role as a diagnostic and prognostic biomarker for ESCC. Citation Format: Mohammad Hussain Hamrah. Secretogranin V as a Potential Biomarker for Esophageal Squamous Cell Carcinoma [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Tumor Immunology and Immunotherapy; 2023 Oct 1-4; Toronto, Ontario, Canada. Philadelphia (PA): AACR; Cancer Immunol Res 2023;11(12 Suppl):Abstract nr A033.

ASF1B acted as a prognostic biomarker for stomach adenocarcinoma.

Stomach adenocarcinoma (STAD) has a high mortality rate due to the lack of highly sensitive biomarkers. Therefore, the search for potential tumor markers is of great value. ASF1B is a prognostic marker for a variety of tumors, while the prognostic value and immune microenvironment of ASF1B in STAD remain unclear, and to be determined. Kaplan-Meier analysis was performed to analyze the prognostic role of ASF1B in STAD. Functional enrichment of ASF1B was explored with GO and KEGG pathway analysis. We also explored the correlation between ASF1B expression and immune infiltration in STAD. ASF1B was significantly upregulated in STAD tissues and high expression of ASF1B indicated a poor overall survival, progression-free survival, and first progression rate in STAD. The functional enrichment analysis of ASF1B and related genes showed high enrichment in the cell cycle and DNA repair, and the ASF1B high expression group was also mainly enriched in pathways such as the cell cycle. Analysis of tumor immune infiltration showed that ASF1B expression was significantly associated with the majority of immune cell infiltration in STAD. Moreover, STAD patients with high ASF1B expression had a higher tumor mutation burden score, microsatellite instability score, PD-1 immunophenoscore, and immune checkpoint expression. Our results suggest that ASF1B was an independent prognostic factor for STAD as well as a potential target for immunotherapy.

Clinical significance of WT1 in the evaluation of therapeutic effect and prognosis of non-M3 acute myeloid leukemia

ABSTRACT To explore the clinical significance and prognosis of acute myeloid leukemia (AML) patients with WT1 mutations.In total, the clinical data of 269 adult patients with non-M3 AML were considered retrospectively. From these patients, 153 carried WT1 mutation whereas 116 were negative. WT1 mutation positive patients were further divided into WT1 low expression and high expression groups base on the expression level of WT1 by qPCR at diagnosis (cut off: 170500). Survival and therapeutic effect analysis were performed for the above patients with different interfering factors such as co-mutations, the extent of WT1 log reduction and the chemotherapy regimens. Patients with high WT1 expression have higher rate of relapse. We can accurately identify patients with inferior outcomes when we take the following factors into consideration: the WT1 expression level at diagnosis; different prognostic factors including co-mutations (especially NPM1 and FLT3-ITD); the log reduction of WT1 after induction therapy and the risk of stratification. Idarubicin + Cytarabine (IA) regimen could reduce the expression level of WT1 after treatment, and Allo-HSCT played an important role in improving the prognosis of patients with WT1 high expression and patients with WT1 negativity. Among the relapsed patients, there existed a rising trend of WT1-MRD in advance than MFC-MRD and that of patients with continuous complete remission (CR). Different clinical background should be taken into consideration when we judge the prognosis and therapeutic effect of patients with WT1 mutations. In addition, WT1 may be an optional MRD marker, which needs regular monitoring.

PBRM1 mutation and WDR72 expression as potential combinatorial biomarker for predicting the response to Nivolumab in patients with ccRCC: a tumor marker prognostic study.

Immune checkpoint therapy (ICT) provides a new idea for the treatment of advanced clear cell renal cell carcinoma (ccRCC), which can bring significant benefits to patients. However, the clinical application of ICT is limited because of the lack of predictive biomarkers to select potential responders. This study aims to propose a new biomarker to predict the response to Nivolumab in patients with ccRCC. The genes that significantly improve the prognosis of ccRCC were retrieved from The Cancer Genome Atlas (TCGA) database. The genomic and clinical data were from patients that had been registered in prospective clinical trials (CheckMate 009, CheckMate 010 and CheckMate 025). TCGA, Gene Expression Omnibus (GEO), and The Human Protein Atlas database were used to analyze the gene and protein expression of WD repeat-containing protein 72 (WDR72) in ccRCC. Gene Ontology (GO) & The Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Set Enrichment Analysis (GSEA) were performed to dig relevant mechanisms of WDR72. Single sample gene set enrichment analysis (ssGSEA) was conducted to evaluate the role of WDR72 in immune infiltration. Cell proliferation assay, FAO and ATP quantification were used to explore and verify the molecular mechanisms. The expression of WDR72, FOXP3, CD8, and CPT1A was examined by IHC in 20 advanced ccRCC tissue samples at the Urology Department of our hospital. The MethSurv was used to identify PBRM1 and WDR72 gene methylation and its effect on prognosis of ccRCC. WDR72 is the most significant gene for improving overall survival (OS) in ccRCC. In all three checkmates, OS and progression free survival (PFS) were found to be significantly higher in WDR72 high expression group than that in WDR72 low expression group (P=0.040 and P=0.012, respectively), and similar conclusions could be drawn from the PBRM1-mutation (MUT) compared with the PBRM1-wildtype (WT) (P=0.007 and P=0.006, respectively). What's more, high expression of WDR72 plus PBRM1-MUT as a combinatorial biomarker showed improved OS (HR=0.388, P=0.0026) and PFS (HR=0.39, P=0.0066) compared to low expression of WDR72 plus PBRM1-WT. Functional enrichment analysis showed that WDR72 was closely positively related to fatty acid degradation and fatty acid beta oxidation pathway in ccRCC. In vitro experiments showed that high expression of WDR72 can promote fatty acids oxidation and inhibit the proliferation of ccRCC cells. Immune analysis revealed that WDR72 high expression was associated with decreased infiltration of Treg cells and low ssGSEA score of check-point. IHC results showed that WDR72 was negatively correlated with FOXP3 expression (r=-0.506, P=0.023) and positively correlated with CPT1A expression (r=0.529, P=0.017). The present study indicated that high expression of WDR72 may indicate a good prognosis of patients treated with Nivolumab and WDR72 expression combined with PBRM1 mutation could be more persuasive to predict the response for ICT in ccRCC patients.

High PD-L1 Expression May be Associated with Worse Overall Prognosis in Acute Myeloid Leukemia

Background: PD-L1 expression has been explored in solid tumours and in classical Hodgkin's lymphoma, however its expression in acute leukemias has been minimally studied. PD-L1 over-expression in AML blasts has previously been associated with worse outcomes in patients with mutated FTL3 genes, and positively correlated with TP53 expression. Over-expression of PD-L1 may contribute to failed response to hypo-methylating agents, leading to the rationale behind combination therapy with immune-checkpoint inhibitors. Despite evidence that the PD-1/PDL-1 axis may promote myeloid neoplasms by modulating immune escape, clinical trials studying the role of immunotherapy in MDS/AML have produced disappointing results. The lack of evidence to support the role of PD-1-targeted therapy in AML may be due to lack of identification of the subset of patients who may benefit from immunotherapy. We previously demonstrated that PD-L1 expression is significantly enhanced in patients with JAK2/STAT mutations compared to patients with wild-type (WT) JAK2/STAT. Here, we investigate the clinical correlate of our findings and compare clinical outcomes of AML patients with respect to PD-L1 expression. Methods: Immunohistochemical staining for PD-L1 was previously performed on bone marrow biopsies of 31 AML patients, including 11 patients with JAK2/STAT mutations (nine with JAK2 V617F exon 14 mutations, one with JAK2 I540-E543delinsRG exon 12 mutation and one with STAT5B N642H mutation) and 20 patients with WT JAK2/STAT. PD-L1 CPS was calculated by summation of all PD-L1-stained cells divided by total number of tumour cells, and multiplied by 100. Retrospective chart review was performed to extract data regarding patient demographics, cytogenetics, mutations, treatment course, as well as clinical outcomes. Low and high PD-L1 expression was defined as CPS score &amp;lt;10 and &amp;gt; 20 respectively. Kaplan-Meier analysis was used for analysing overall survival (OS) and event-free survival (EFS), with statistical significance determined using a 2-tailed log-rank test. EFS was defined as duration of time between diagnosis and the first occurrence of either going off protocol induction without achieving complete remission (CR), relapse from CR, or death due to any cause. Patients who were alive at the time of study analysis were censored at date of last follow-up. Results: A total 15 patients demonstrated high PD-L1 expression (lowest PD-L1 CPS 20), including four patients with WT JAK2/STAT and all 11 patients with mutant JAK2/STAT. 16 patients demonstrated low PD-L1 expression (highest PD-L1 CPS 6). Only three patients in the low PD-L1 expression group never achieved complete remission compared to eight patients in the high PD-L1 expression group. Median OS in the low PD-L1 expression group was significantly longer than median OS in the high PD-L1 expression group (29.1 months [95% CI 1.4-7.2] vs 9.3 months [95% CI 0.1-0.7]; HR 2.4 [95% CI 1.1-5.6], p = 0.0253, Figure 1). Similarity, EFS was significantly longer in the low PD-L1 expression group (13.1 months [95% CI 1.3-6.7] vs. 4.4 months [95% CI 0.1-0.8]; HR 2.4 [95% CI 1.1-5.6], p = 0.0253). When analysis was separated by European LeukemiaNet (ELN) risk stratification, significant differences in OS and EFS were re-demonstrated in patients with favourable ELN risk, but not in patients with intermediate or adverse risks. When wild-type JAK2/STAT patients were analyzed separately, there was no significant differences between OS and EFS with respect to PD-L1 expression. Conclusions: We previously demonstrated that PD-L1 is significantly up-regulated in AML patients who carry mutant JAK2/STAT when compared to WT JAK2. Using a proposed PD-L1 CPS cut-off score of 20, we demonstrated that high PD-L1 expression was associated with significantly worse outcomes with respect to OS and EFS. These results may indicate a therapeutic role of immunotherapy in the treatment of AML, particularly in patients with JAK2/STAT mutations.

Assessment of Ki-67 proliferation index in prognosis prediction in patients with nonmetastatic clear cell renal cell carcinoma and tumor thrombus

PurposeTo determine the optimal cut-off value of Ki-67 for predicting the survival of patients with clear cell renal cell carcinoma (ccRCC) and tumor thrombus and to explore the correlation between Ki-67 expression and pathological features. Patients and MethodsWe retrospectively analyzed Ki-67 immunohistochemical staining of ccRCC and tumor thrombus resected from February 2006 to February 2022. The survival rate was evaluated using the Kaplan-Meier method. The optimal cut-off value of the Ki-67 expression for predicting survival was determined by the minimum P-value method. Clinicopathological data were compared based on Ki-67 status (low versus high expression). Univariate and multivariate Cox regression analysis was used to explore independent predictors. ResultsA total of 202 patients (median age, 58 years [IQR, 52–65 years], 147 men) with ccRCC and tumor thrombus were included in the study. The optimal cut-off value of Ki-67 for predicting survival was 30%. 159 (78.7%) and 43 (21.3%) patients were included in the low-expression and high-expression groups. Patients with Ki-67 high expression had significantly worse recurrence-free survival (P < 0.001) and cancer-specific survival (P < 0.001). Ki-67 high expression was associated with adverse pathological features, including tumor necrosis, ISUP nuclear grade, sarcomatoid differentiation, perirenal fat invasion, renal pelvis invasion, and inferior vena cava wall invasion (all P < 0.050). Ki-67 expression ≥ 30% (P = 0.016), tumor side (P = 0.003), diabetes (P = 0.040), blood loss (P = 0.016), inferior vena cava wall invasion (P = 0.016), and sarcomatoid differentiation (P = 0.014) were independent predictors of cancer-specific survival. ConclusionThe optimal cut-off level of Ki-67 in predicting the prognosis of ccRCC and tumor thrombus was 30%. The high expression of Ki-67 was associated with the aggressive pathological phenotype and poor prognosis.

Targeting Myeloid Epithelial Tyrosine Kinase (MERTK) Receptor in Acute Myeloid Leukemia Using a Novel Antibody Drug Conjugate, Rgx-019-MMAE

Background: Myeloid epithelial reproductive tyrosine kinase receptor (MERTK) is a transmembrane protein receptor from the TAM (Tyro3, Axl, Mertk) family. It is known to be overexpressed in various solid and hematological cancers. Overexpression of MERTK is associated with reduced apoptosis, increased metastasis, and drug resistance in tumor cells, making it an attractive therapeutic target. RGX-019-MMAE, a novel humanized IgG1-MMAE antibody-drug conjugate (ADC) developed by Inspirna, selectively binds to MERTK on tumor cells, which results in internalization and degradation of the receptor, and induces cytotoxicity through the release of the payload, MMAE, which disrupts the mitosis process of actively dividing cells. Several monoclonal antibodies targeting MERTK have shown promising results against various solid cancers, however, the effect of targeting MERTK in AML progression via RGX-019-MMAE is not known. Here, we hypothesize MERTK is overexpressed in distinct subsets of AML and associated with leukemia progression, therefore, targeting MERTK via RGX-019-MMAE exerts an antileukemic effect. Methods: To determine the clinical/prognostic relevance of MERTK, we analyzed a gene expression dataset (OHSU) and segregated the patients into low and high MERTK mRNA expression groups based on their mean expression value. To identify the specific AML subset with the highest MERTK expression, MERTK protein expression was analyzed in 818 patients with AML by performing Reverse Phase Protein Arrays (RPPA). MERTK protein expression was also measured by flow cytometry in OCI-AML3, OCI-AML2, Kasumi-1, ThP-1, Molm-13, Molm-14, MV4-11, and U937 leukemic cell lines and in the peripheral blood or bone marrow of five AML patients. Further, we treated Kasumi-1 and OCI-AML3 AML cell lines with varying doses of RGX-019-MMAE or isotype control for 72 and 120 hours and measured its effect on AML cell viability using CellTiterGlo 2.0 assay. Likewise, we treated primary cells from 5 different AML patients and determined the anti-leukemic effect of RGX-019-MMAE in primary cells. Finally, we investigated the synergistic effect of RGX-019-MMAE with Venetoclax (BCL2 inhibitor) or 5-Azacytidine in vitro. Results: Analysis of AML OHSU dataset showed that patients with high MERTK mRNA expression had significantly worse overall survival than those having low MERTK mRNA expression (p=0.02). RPPA analysis revealed significantly higher MERTK protein expression in monocytic AML subtypes, such as M4 and M5, especially in those with PTPN11 mutation or t (9;11) translocation. Moreover, in the RPPA we noted that high expression of MERTK protein levels is significantly correlated with inferior survival. This finding suggests that MERTK could serve as a therapeutic target in AML, especially in patients with monocytic subtypes of leukemia. In addition, we observed varying degrees of MERTK protein expression in AML cell lines, with Kasumi-1 and OCI-AML3 expressing the highest amount of MERTK protein. Next, treatment of Kasumi-1 and OCI-AML3 cell lines with varying doses of anti-MERTK ADC, RGX-019-MMAE, resulted in significantly more killing of leukemic cells than the isotype control ADC in a dose-dependent manner (p&amp;lt;0.01), suggesting that RGX-019-MMAE inhibits AML cell proliferation. Further, we validated this finding in primary AML cells with high MERTK expression. As expected, we observed that treatment with RGX-019-MMAE induced significantly greater cell death (˜80% apoptosis) in leukemic cells than the isotype control ADC. In addition, Kasumi-1 and OCl-AML3 cells treated in combination with RGX-019-MMAE and either Venetoclax or 5-Azacytidine showed a synergistic effect with increased cytotoxicity in a dose-dependent manner. These findings suggest that RGX-019-MMAE targets MERTK-expressing leukemic cells, sensitizes cells to chemotherapies, and promotes overall cytotoxicity. Conclusion: RGX-019-MMAE significantly induced cell death in AML cell lines and primary AML patient samples. RGX-019-MMAE, combined with chemotherapeutic agents and targeted therapy, produces a synergistic anti-leukemic effect. Our data indicates that MERTK is a potential therapeutic target in AML patients with monocytic subtypes of leukemia.

Prognostic and Therapeutic Implications of TP53 expression in Chronic Myelomonocytic Leukaemia: Results of a Multicentre Study

Thetumor suppressor TP53 is among the most mutated genes in cancer, including many hematological malignancies in which it usually imparts more aggressive, chemo-refractory, and poor prognosis diseases. TP53 mutations are, though, remarkably rare in chronic myelomonocytic leukemia (CMML): affecting 2.4% of cases in a recent study ( Gurney et al, Leukemia 2023). We found pathogenic TP53 mutations ( TP53MT) in only 1.1% of 640 genotyped UK CMML patients. As expected, TP53MT conferred worse leukemia-free (LFS: median 23 v 28 mo, p=0.031) and overall (OS: 26 v 29 mo, p=0.015) survival compared with TP53WTcases. The clinical relevance in CMML of TP53 expression, however, remains unexplored. To address this we studied 92 TP53WT CMML patients (discovery cohort) from the National Taiwan University Hospital (Taipei, Taiwan) with available RNA-seq on presentation bone marrow (BM), each with extensive longitudinal clinical, laboratory and outcome data. Validation cohorts comprised 33 RNA-sequenced patients from The Christie (Manchester, UK) and 24 from Hospital Morales Meseguer (Murcia, Spain). For all cohorts, RNA-seq data from healthy controls (HCs) were available. Patients in the discovery cohort were divided into low and high TP53 expression groups, determined by the maximally selected rank statistics. TP53low patients (n=15) displayed significantly lower expression than HCs, while TP53high (n=77) had expression levels comparable to HCs. Thus, although most CMML patients are TP53WT, a subset has down-regulated TP53 expression and potential for altered p53 (and downstream) functions. TP53low and TP53high patients did not differ in clinical features or concurrent genetic alterations, although in all cohorts TP53low patients showed a trend towards poorer response to hypomethylating agents (HMA). TP53 low patients displayed significantly worse outcomes than TP53high patients (LFS 7 v 19 mo, p=0.002; OS 11 v 26 mo, p=0.001). Moreover, TP53 expression further stratified prognosis within different CMML-specific prognostic scoring system (CPSS)-molecular risk groups (Fig 1) . Time-dependent ROC analysis indicated TP53 expression as complementary to current risk stratification systems. In multivariable analysis, low TP53 expression remained prognostically detrimental. Survival analyses were consistent in both validation cohorts. GSEA and IPA analyses revealed that compared with HCs or TP53high, TP53low cells exhibited depleted expression of p53-dependent pathways, including MYC targets, E2F targets, G2/M checkpoints, and DNA repair. Interestingly, these are all among the most upregulated pathways in TP53MT (v TP53WT) samples across multiple cancers in TCGA data ( Donehower et al, Cell Rep 2019), indicating distinct roles for p53 in regulating these pathways in a TP53WT context, and that the driving biology of TP53low CMML is distinct from (and not functionally equivalent to) that of oncogenic TP53 mutations. TP53low patients demonstrated significantly enhanced TNF-alpha and inflammatory response signals, highlighting other distinctive pathobiology of the TP53low subgroup. In line with our clinical observation, single-sample GSEA showed significant enrichment of HMA resistance signatures in TP53low patients, potentially contributing to the dismal prognosis in this group. We next investigated whether this could be exploited therapeutically. We treated primary CMML BM cells ex vivo with azacitidine and NSC-207895, an MDMX inhibitor with p53 activating properties, observing clear and substantial synergy for combination therapy in samples from 10 of 11 patients. There was a trend towards inverse correlation between TP53 expression and empirical synergy scores (r=-0.35), suggesting potential for p53 activation to enhance HMA sensitivity in CMML, with broad efficacy, perhaps preferentially in adverse risk TP53low cases. In summary, we have identified and validated, across three cohorts from three countries, that a subgroup of CMML patients display aberrantly low TP53 expression, associated with HMA resistance and worse survival. Transcriptomic analyses revealed distinctive biology driving this novel TP53low subgroup (Fig 2), mandating further mechanistic study. Importantly we report consistent and substantial synergy for combining pharmacological p53 activation with HMA, presenting a novel targeted approach to improve HMA response for this unmet clinical need.

Complete pathological remission and tertiary lymphoid structures are associated with the efficacy of resectable NSCLC receiving neoadjuvant chemoimmunotherapy: A double-center retrospective study

ABSTRACT This study aimed to investigate the relationship between complete pathological remission (PCR), tertiary lymphoid structure (TLS) maturation and expression and clinical outcomes of patients with resectable non-small cell lung cancer (NSCLC) receiving neoadjuvant chemoimmunotherapy. Totally 80 patients with resectable NSCLC (stage IB-IIIB) receiving neoadjuvant chemoimmunotherapy were analyzed. We used the Kaplan-Meier method to plot survival curves and the log-rank test to compare differences. Among all patients included, 45 patients (56.25%) achieved major pathological response (MPR), including 30 patients (37.50%) with PCR. The proportion of patients diagnosed with stage IB, II, IIIA and IIIB was 1.25%, 10.00%, 52.50% and 36.25%, respectively. We divided patients into PCR group and non-PCR group respectively according to whether they achieved PCR. We found that patients achieving PCR had significantly improved disease-free survival (DFS) (mDFS: NR vs. 20.24 months, P = .020). TLS expression was low in 43 cases (53.75%) and high in 37 cases (46.25%). TLS maturation was low in 55 cases (68.75%) and high in 25 cases (31.25%). The DFS of patients with TLS high-maturation (34.07 vs. 22.30 months, P = .024) and TLS high-expression (34.07 vs. 22.30 months, P = .041) was significantly longer. In most subgroups, the PCR, TLS high-maturation and TLS high-expression group respectively achieved a better clinical outcome relative to the non-PCR, TLS low-maturation and TLS low-expression group. In patients with resectable NSCLC receiving neoadjuvant chemoimmunotherapy, the acquirement of PCR may predict better DFS. In addition, high expression and maturation of TLS may be prognostic factors.

PBK correlates with prognosis, immune escape and drug response in LUAD

PBK (PDZ-binding kinase) is a protein-coding gene that encodes a serine/threonine protein kinase associated with the dual-specific mitogen-activated protein kinase (MAPKK) family. Overexpression of this gene is closely linked to tumor development. In this study, we aimed to investigate the role of PBK in lung adenocarcinoma (LUAD) progression, prognosis, and immune evasion. We conducted a pan-cancer analysis of PBK to examine its expression and prognostic value. In the LUAD cohort, we analyzed PBK expression, prognosis, mutational features, and immune infiltration in groups with different PBK expression levels. We constructed a PBK-associated genomic model, integrated it into a nomogram, and compared high and low-risk subgroups. In our pan-cancer analysis, PBK was significantly upregulated, particularly in LUAD patients, and displayed poor prognosis. The high PBK expression group had many deletion mutations but still showed gene upregulation. Immune infiltration analysis indicated that PBK-triggered immune escape in the high expression group might relate to antigen presentation, dendritic cell, and CD8+ T cell infiltration. We constructed a 5-gene prognostic model and a nomogram to quantify individual survival probabilities. The PBK-associated gene prognostic model reliably predicted patient prognosis and drug response. Our findings offer new insights into PBK-induced immune escape and targeted therapy during LUAD development, providing valuable suggestions for clinical treatment approaches.

Vasari-Based Features Nomogram to Predict the Tumor-Infiltrating CD8+ T Cell Levels in Glioblastoma

Tumor-infiltrating CD8+T cells play a key role inglioblastoma (GB) development, malignant progression, and recurrence. The aim of the study was to establish nomograms based on the Visually AcceSAble Rembrandt Images (VASARI) features of multiparametric magnetic resonance imaging (MRI) to determine the expression levels of tumor-infiltrating CD8+T cells in patients with GB. Pathological and imaging data of 140 patients with GB confirmed by surgery and pathology were retrospectively analyzed. The levels of tumor-infiltrating CD8+T cells in tumor tissue samples obtained from patients were quantified using immunohistochemical staining. Patients were divided into high and low CD8 expression groups. The MRI images of patients with GBwere analyzed by two radiologists using the VASARI scoring system. A total of 25 MRI-based VASARI imaging features were evaluated by two neuroradiologists. The features with the greatest predictive power for CD8 expression levels were, cystic (OR, 3.063; 95% CI: 1.387, 6.766; P=0.006), hemorrhage (OR, 2.980; 95% CI: 1.172, 7.575; P=0.022), and ependymal extension (OR, 0.257; 95% CI: 0.114 0.581; P=0.001). A logistic regression model based on these three features showed better sample predictive performance (AUC=0.745; 95% CI: 0.665, 0.825; Sensitivity=0.527; Specificity=0.857). The VASARI feature-based nomogram model can showpromise to predict the level of infiltrative CD8 expression in GB tumors non-invasively for earlier tissue diagnosis and more aggressive treatment.

Integrating scRNA-seq to explore novel macrophage infiltration-associated biomarkers for diagnosis of heart failure

ObjectiveInflammation and immune cells are closely intertwined mechanisms that contribute to the progression of heart failure (HF). Nonetheless, there is a paucity of information regarding the distinct features of dysregulated immune cells and efficient diagnostic biomarkers linked with HF. This study aims to explore diagnostic biomarkers related to immune cells in HF to gain new insights into the underlying molecular mechanisms of HF and to provide novel perspectives for the detection and treatment of HF.MethodThe CIBERSORT method was employed to quantify 22 types of immune cells in HF and normal subjects from publicly available GEO databases (GSE3586, GSE42955, GSE57338, and GSE79962). Machine learning methods were utilized to screen for important cell types. Single-cell RNA sequencing (GSE145154) was further utilized to identify important cell types and hub genes. WGCNA was employed to screen for immune cell-related genes and ultimately diagnostic models were constructed and evaluated. To validate these predictive results, blood samples were collected from 40 normal controls and 40 HF patients for RT-qPCR analysis. Lastly, key cell clusters were divided into high and low biomarker expression groups to identify transcription factors that may affect biomarkers.ResultsThe study found a noticeable difference in immune environment between HF and normal subjects. Macrophages were identified as key immune cells by machine learning. Single-cell analysis further showed that macrophages differed dramatically between HF and normal subjects. This study revealed the existence of five subsets of macrophages that have different differentiation states. Based on module genes most relevant to macrophages, macrophage differentiation-related genes (MDRGs), and DEGs in HF and normal subjects from GEO datasets, four genes (CD163, RNASE2, LYVE1, and VSIG4) were identified as valid diagnostic markers for HF. Ultimately, a diagnostic model containing two hub genes was constructed and then validated with a validation dataset and clinical samples. In addition, key transcription factors driving or maintaining the biomarkers expression programs were identified.ConclusionThe analytical results and diagnostic model of this study can assist clinicians in identifying high-risk individuals, thereby aiding in guiding treatment decisions for patients with HF.