Journal: Cancers

Article Title: An Integrated Immunometabolic Signature Predicts Prognosis and Immunotherapy Response in ccRCC and Identifies UCN -Mediated Immune Evasion as a Therapeutic Vulnerability: Evidence from In Vitro and In Vivo Studies

doi: 10.3390/cancers18091373

Figure Lengend Snippet: The process of identifying differentially expressed IMRGs and molecular subtypes in ccRCC: ( A ) IMRGs that were differentially expressed were denoted by red dots for upregulation and blue dots for downregulation. ( B ) Heatmaps were used to visually represent the top differentially expressed genes. ( C ) A heatmap of the nsNMF consensus matrix was generated to classify ccRCC into two molecular subtypes. ( D ) A PCA plot was applied to show significant differences between clusters. ( E ) The gene expression heatmap shows how the identified IMRGs were expressed across the two molecular subtypes. ( F , G ) In order to make a comparison between the two molecular subtypes, the researcher employed the Kaplan–Meier curve to assess and contrast the OS and PFS.

Article Snippet: Human ccRCC cell line 786-O (Accession Number: CVCL_1051) and mouse ccRCC cell line Renca (CVCL_2174) were obtained from American Type Culture Collection (ATCC) (Manassas, Virginia) and cultured in RPMI 1640 medium (Procell, Wuhan, China) containing 10% fetal bovine serum (Procell, China) and Penicillin–Streptomycin (Procell, China).

Techniques: Generated, Gene Expression, Comparison

Journal: Cancers

Article Title: An Integrated Immunometabolic Signature Predicts Prognosis and Immunotherapy Response in ccRCC and Identifies UCN -Mediated Immune Evasion as a Therapeutic Vulnerability: Evidence from In Vitro and In Vivo Studies

doi: 10.3390/cancers18091373

Figure Lengend Snippet: Comparison of genomic alteration landscapes between the two molecular subtypes. ( A ) Oncoplot demonstrated the 30 most frequently mutated genes in Cluster 1. ( B ) Oncoplot demonstrated the 30 most frequently mutated genes in Cluster 2. ( C ) Heatmap illustrating the co-mutated states of the commonly mutated genes in Cluster 1. ( D ) Heatmap illustrating the co-mutated states of the commonly mutated genes in Cluster 2. ( E ) The boxplot illustrates the distinct tumor mutation frequencies between Cluster 1 and Cluster 2. ( F ) The Kaplan–Meier curve shows the overall survival rates of patients with high and low tumor mutation burdens. ( G ) Multivariate Cox regression analysis of tumor mutation burden (TMB) and immunometabolic clusters. ( H ) Kaplan–Meier survival curves for ccRCC patients stratified by both TMB status (high vs. low) and immunometabolic clusters (C1 vs. C2).

Article Snippet: Human ccRCC cell line 786-O (Accession Number: CVCL_1051) and mouse ccRCC cell line Renca (CVCL_2174) were obtained from American Type Culture Collection (ATCC) (Manassas, Virginia) and cultured in RPMI 1640 medium (Procell, Wuhan, China) containing 10% fetal bovine serum (Procell, China) and Penicillin–Streptomycin (Procell, China).

Techniques: Comparison, Mutagenesis

Journal: Cancers

Article Title: An Integrated Immunometabolic Signature Predicts Prognosis and Immunotherapy Response in ccRCC and Identifies UCN -Mediated Immune Evasion as a Therapeutic Vulnerability: Evidence from In Vitro and In Vivo Studies

doi: 10.3390/cancers18091373

Figure Lengend Snippet: Assessment and confirmation of the predictive performance of the signature in ccRCC. ( A – C ) Scatter plots illustrating the survival status and IMI scores of ccRCC patients in the TCGA training group ( A ), the TCGA testing group ( B ), and the E-MATB-1980 external validation group ( C ). ( D – F ) Kaplan–Meier curves displaying the overall survival situation per IMI scores of the high-IMI group and low-IMI group in the TCGA training group ( D ), the TCGA testing group ( E ), and the E-MATB-1980 external validation group ( F ). ( G – I ) ROC curves demonstrating the predictive performance of IMI with AUC values for 1-year, 3-year, and 5-year OS in ccRCC patients from the TCGA training group ( G ), the TCGA testing group ( H ), and the E-MATB-1980 external validation group ( I ).

Article Snippet: Human ccRCC cell line 786-O (Accession Number: CVCL_1051) and mouse ccRCC cell line Renca (CVCL_2174) were obtained from American Type Culture Collection (ATCC) (Manassas, Virginia) and cultured in RPMI 1640 medium (Procell, Wuhan, China) containing 10% fetal bovine serum (Procell, China) and Penicillin–Streptomycin (Procell, China).

Techniques: Biomarker Discovery

Journal: Cancers

Article Title: An Integrated Immunometabolic Signature Predicts Prognosis and Immunotherapy Response in ccRCC and Identifies UCN -Mediated Immune Evasion as a Therapeutic Vulnerability: Evidence from In Vitro and In Vivo Studies

doi: 10.3390/cancers18091373

Figure Lengend Snippet: Identification of expression trends of nine IMRGs. ( A ) Differences in signature gene expression between high and low IMI groups in the TCGA database. ns, not significant; * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001. ( B ) Differences in signature gene expression between normal kidney tissue samples and ccRCC samples in the TCGA database. ( C – K ) The relative expression levels of signature genes between three ccRCC cell lines (786-O, A498, ACHN) and normal renal tubular epithelial cells, HK2. ( L ) The IHC images compared the expression levels of four signature genes between normal renal tissue samples and ccRCC samples in the HPA database ( https://www.proteinatlas.org , accessed on 1 January 2024).

Article Snippet: Human ccRCC cell line 786-O (Accession Number: CVCL_1051) and mouse ccRCC cell line Renca (CVCL_2174) were obtained from American Type Culture Collection (ATCC) (Manassas, Virginia) and cultured in RPMI 1640 medium (Procell, Wuhan, China) containing 10% fetal bovine serum (Procell, China) and Penicillin–Streptomycin (Procell, China).

Techniques: Expressing, Gene Expression

Journal: Cancers

Article Title: An Integrated Immunometabolic Signature Predicts Prognosis and Immunotherapy Response in ccRCC and Identifies UCN -Mediated Immune Evasion as a Therapeutic Vulnerability: Evidence from In Vitro and In Vivo Studies

doi: 10.3390/cancers18091373

Figure Lengend Snippet: Verification of UCN promoting proliferation, migration, and invasion of ccRCC. ( A ) Knockdown of the UCN gene in 786-O and ACHN cells, relative mRNA levels in the negative control (NC) group and three siRNA knockdown groups, respectively. **** p < 0.0001 ( B ) The knockdown effect of three siRNAs on the UCN gene at the protein level in two cell lines. The uncropped blots are shown in . ( C ) The proliferation curves of CCK8 in the control group and the knockdown groups of the two cell lines. Any siRNA group has significant statistical differences from the NC group. ( D , E ) Wound-healing assays in control and knockdown groups of the two cell lines. ( F , G ) Transwell invasion assays in control and knockdown groups of the two cell lines.

Article Snippet: Human ccRCC cell line 786-O (Accession Number: CVCL_1051) and mouse ccRCC cell line Renca (CVCL_2174) were obtained from American Type Culture Collection (ATCC) (Manassas, Virginia) and cultured in RPMI 1640 medium (Procell, Wuhan, China) containing 10% fetal bovine serum (Procell, China) and Penicillin–Streptomycin (Procell, China).

Techniques: Migration, Knockdown, Negative Control, Control

Journal: Cancers

Article Title: An Integrated Immunometabolic Signature Predicts Prognosis and Immunotherapy Response in ccRCC and Identifies UCN -Mediated Immune Evasion as a Therapeutic Vulnerability: Evidence from In Vitro and In Vivo Studies

doi: 10.3390/cancers18091373

Figure Lengend Snippet: UCN regulates the immune microenvironment and promotes ccRCC progression. ( A ) Schematic illustration of the mouse xenograft tumor model experimental design. ( B – E ) Tumor growth analyses demonstrate reduced tumor volume and weight across different experimental groups, with notable suppression in sh UCN +IgG2a and sh UCN +PD-1 groups. ( F ) Gating strategy for tumor-infiltrating lymphocytes. Representative flow plots showing the identification of Live/CD45+ cells, T cells (CD3+), CD4+ and CD8+ subsets, as well as Tregs and PD-1+ cells. ( G ) Flow cytometry analysis unveils substantial alterations in immune cell subsets in the tumor immune microenvironment. ( H , I ) Representative mIHC staining of tumors (green: CD8, red: Foxp3, blue: DAPI; scale bar, 50 μm.) ( I ) The column diagram showing the counts of spots with CD8+ T cells and Tregs in tumor slides. Data presented as Mean ± SEM. One-way ANOVA was used in ( E , G , I ). * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001.

Article Snippet: Human ccRCC cell line 786-O (Accession Number: CVCL_1051) and mouse ccRCC cell line Renca (CVCL_2174) were obtained from American Type Culture Collection (ATCC) (Manassas, Virginia) and cultured in RPMI 1640 medium (Procell, Wuhan, China) containing 10% fetal bovine serum (Procell, China) and Penicillin–Streptomycin (Procell, China).

Techniques: Flow Cytometry, Staining

Journal: Molecular Cancer Therapeutics

Article Title: Targeted Delivery of a Potent STING Agonist Payload via an Antibody–Drug Conjugate Drives Robust Antitumor Activity in Preclinical Models

doi: 10.1158/1535-7163.MCT-25-0108

Figure Lengend Snippet: Tumor-targeted ncSTING ADCs drive antitumor activity across multiple tumor models with multiple tumor antigens. Mean tumor volume over time of ( A ) human CD228-LL2 tumor–bearing C57BL/6 mice, ( B ) murine B7-H4-EMT6 tumor–bearing C57BL/6 mice, ( C ) murine B7-H4-Renca tumor–bearing Balb/c mice, and ( D ) murine IB6-CT26 tumor–bearing Balb/c mice following treatment with three weekly intraperitoneal doses (arrows) of the indicated ADCs. Data in each panel represent a single in vivo experiment. Following a one-way ANOVA test, a Tukey post hoc multiple comparisons test was used to compare each treatment group: ****, P < 0.0001; ***, P < 0.001; **, P < 0.01; *, P < 0.05. Error bars depict SD.

Article Snippet: Renca tumor cells (ATCC, #CRL-2947, RRID:CVCL_2174; purchased in 2004) were cultured in RPMI 1640 medium (ATCC) with 10% heat-inactivated FBS, MEM nonessential amino acids (0.1 mmol/L), sodium pyruvate (1 mmol/L), and L-glutamine (2 mmol/L) and implanted subcutaneously (2 × 10 6 cells in 200 μL 25% Matrigel in RPMI 1640) into Balb/c female mice (RRID:IMSR_ENV:HSD-047).

Techniques: Activity Assay, In Vivo

Journal: Molecular Cancer Therapeutics

Article Title: Targeted Delivery of a Potent STING Agonist Payload via an Antibody–Drug Conjugate Drives Robust Antitumor Activity in Preclinical Models

doi: 10.1158/1535-7163.MCT-25-0108

Figure Lengend Snippet: Tumor-targeted STING agonist delivery via an ADC minimizes systemic cytokine induction compared with systemic delivery of the released payload. A and C, Mean tumor volume over time of Renca tumor–bearing Balb/c mice following treatment with three weekly intravenous ( A ) or intraperitoneal ( C ) doses (arrows) of the indicated ADCs or released payload. B and D, Quantification of IL-6 in the plasma 3, 6, and 24 hours following treatment as in A and C . Data in each panel represent a single in vivo experiment. Following a one-way ANOVA test, a Tukey post hoc multiple comparisons test was used to compare AUC.3 values ( A and C ) or cytokine levels ( B and D ) for each treatment group: ****, P < 0.0001; ***, P < 0.001; **, P < 0.01; *, P < 0.05. Error bars depict SD.

Article Snippet: Renca tumor cells (ATCC, #CRL-2947, RRID:CVCL_2174; purchased in 2004) were cultured in RPMI 1640 medium (ATCC) with 10% heat-inactivated FBS, MEM nonessential amino acids (0.1 mmol/L), sodium pyruvate (1 mmol/L), and L-glutamine (2 mmol/L) and implanted subcutaneously (2 × 10 6 cells in 200 μL 25% Matrigel in RPMI 1640) into Balb/c female mice (RRID:IMSR_ENV:HSD-047).

Techniques: Clinical Proteomics, In Vivo

Journal: Molecular Cancer Therapeutics

Article Title: Targeted Delivery of a Potent STING Agonist Payload via an Antibody–Drug Conjugate Drives Robust Antitumor Activity in Preclinical Models

doi: 10.1158/1535-7163.MCT-25-0108

Figure Lengend Snippet: Tumor-targeted ncSTING ADCs with a WT Fc backbone drive enhanced antitumor activity in some, but not all, tumor models. Mean tumor volume over time of ( A ) human CD228-LL2 tumor–bearing C57BL/6 mice, ( B ) murine B7-H4-EMT6 tumor–bearing C57BL/6 mice, ( C ) murine B7-H4-Renca tumor–bearing Balb/c mice, ( D ) murine IB6-CT26 tumor–bearing Balb/c mice, ( E ) MC38 tumor–bearing C57BL/6 mice, and ( F ) MC38 tumor–bearing C57BL/6 WT and STING-deficient mice following treatment with the three weekly intraperitoneal doses (arrows) or a single dose (denoted by “x1”) of the indicated ADCs. Data in each panel represent a single in vivo experiment. Following a one-way ANOVA test, a Tukey post hoc multiple comparisons test was used to compare AUC.3 values for each treatment group: ****, P < 0.0001; ***, P < 0.001; **, P < 0.01; *, P < 0.05. Error bars depict SD.

Article Snippet: Renca tumor cells (ATCC, #CRL-2947, RRID:CVCL_2174; purchased in 2004) were cultured in RPMI 1640 medium (ATCC) with 10% heat-inactivated FBS, MEM nonessential amino acids (0.1 mmol/L), sodium pyruvate (1 mmol/L), and L-glutamine (2 mmol/L) and implanted subcutaneously (2 × 10 6 cells in 200 μL 25% Matrigel in RPMI 1640) into Balb/c female mice (RRID:IMSR_ENV:HSD-047).

Techniques: Activity Assay, In Vivo