Journal: Journal for Immunotherapy of Cancer

Article Title: BTN1A1 is a novel immune checkpoint mutually exclusive to PD-L1

doi: 10.1136/jitc-2023-008303

Figure Lengend Snippet: Summary of butyrophilin 1A1 staining results in tissue microarray scores for patients with different solid and hematological tumor types

Article Snippet: Squamous non-small cell lung cancer , AMSBIO LC1921c , 84% , 10% , 6%.

Techniques: Staining, Microarray

Journal: Journal for Immunotherapy of Cancer

Article Title: BTN1A1 is a novel immune checkpoint mutually exclusive to PD-L1

doi: 10.1136/jitc-2023-008303

Figure Lengend Snippet: Summary of butyrophilin 1A1 staining results in tissue microarray scores for patients with different solid and hematological tumor types

Article Snippet: Squamous non-small cell lung cancer , AMSBIO LC1921c , 84% , 10% , 6%.

Techniques: Staining, Microarray

Journal: Journal for Immunotherapy of Cancer

Article Title: BTN1A1 is a novel immune checkpoint mutually exclusive to PD-L1

doi: 10.1136/jitc-2023-008303

Figure Lengend Snippet: Summary of butyrophilin 1A1 staining results in tissue microarray scores for patients with different solid and hematological tumor types

Article Snippet: Adenocarcinoma non-small cell lung cancer , AMSBIO LC1912c , 45% , 20% , 35%.

Techniques: Staining, Microarray

Journal: Journal for Immunotherapy of Cancer

Article Title: BTN1A1 is a novel immune checkpoint mutually exclusive to PD-L1

doi: 10.1136/jitc-2023-008303

Figure Lengend Snippet: Summary of butyrophilin 1A1 staining results in tissue microarray scores for patients with different solid and hematological tumor types

Article Snippet: Squamous non-small cell lung cancer , AMSBIO LC1921c , 84% , 10% , 6%.

Techniques: Staining, Microarray

Journal: Journal for Immunotherapy of Cancer

Article Title: Rationally targeted anti-VISTA antibody that blockades the C-C’ loop region can reverse VISTA immune suppression and remodel the immune microenvironment to potently inhibit tumor growth in an Fc independent manner

doi: 10.1136/jitc-2021-003382

Figure Lengend Snippet: VISTA is predominantly expressed on myeloid-derived cells in healthy tissues and several solid tumors, including TNBC and NSCLC, show significant expression of VISTA and its binding partner VSIG3. (A) Immune cell cluster identification by RNA single-cell analysis of 10X Genomics data comprising 68,000 healthy PBMCs. (B and C) Uniform Manifold Approximation and Projection (UMAP) and summary plot of single-cell RNA-seq data for VISTA expression in different cell clusters, where cDC refers to classical dendritic cells, pDC to plasmacytoid dendritic cells and HSPC to hematopoietic stem and progenitor cells. (D) Representative immunohistochemical staining for healthy spleen, bone marrow, breast and lung TMAs (n=99) stained with HMBD-002-mIgG2a at 0.02 mg/mL; magnification 200×. (E) Representative immunohistochemical staining for VISTA in TNBC (n=126) and NSCLC TMAs (n=140) stained with HMBD-002-mIgG2a at 0.02 mg/mL; magnification 200× (top) and % of patients with negative (0), low (1), moderate (2) or high (3) staining intensity (bottom). (F) Representative immunohistochemical staining for VSIG3 in TNBC (n=126) and NSCLC TMAs (n=140) stained with anti-VSIG3 at 0.003 mg/mL, magnification 200× (top) and % of patients with negative (0), low (1), moderate (2) or high (3) staining intensity (bottom). HCC, hepatocellular carcinoma; IHC, immunohistochemistry; NSCLC, non-small cell lung cancer; scRNA-seq, single-cell RNA sequencing; TMA, tissue microarrays; TNBC, triple negative breast cancer; VISTA, V-domain Ig suppressor of T cell activation; VSIG3, V-Set and Immunoglobulin domain containing 3.

Article Snippet: Tissue microarrays (TMA) of patients with triple negative breast cancer (TNBC), non-small cell lung cancer (NSCLC), mesothelioma and liver cancer (USBiomax, #BR1301, #LC1401, #MS481d, #LV8013a) comprising formalin-fixed paraffin-embedded (FFPE) tissues were stained for VISTA (HMBD-002-mIgG2a; dilution 1:800) and VSIG3 (LS Biosciences #LS-C338858; dilution 1:300) and normal TMA (USBiomax #FDA999q) was also stained for VISTA (HMBD-002-mIgG2a).

Techniques: Derivative Assay, Expressing, Binding Assay, Single-cell Analysis, RNA Sequencing Assay, Immunohistochemical staining, Staining, Immunohistochemistry, Activation Assay

Journal: Journal of Cellular and Molecular Medicine

Article Title: Identification of natural compounds tubercidin and lycorine HCl against small‐cell lung cancer and BCAT1 as a therapeutic target

doi: 10.1111/jcmm.17246

Figure Lengend Snippet: Clinical relevance of BCAT1 in SCLC. Expression of EIF4G1 in formalin‐fixed paraffin‐embedded (FFPE) SCLC and normal lung tissue arrays were determined using immunohistochemistry (IHC). (A) The IHC images from representative cases. (B–D) The percentage of DAB stained pixels were determined by analysing the raw images with the QuPath software (version 0.2.3). The nested graphs show expressional difference among these groups. ** p < 0.01 (vs the normal lung tissue group, the IA group or the T1 group respectively); ns: not significant

Article Snippet: The SCLC formalin‐fixed, paraffin‐embedded (FFPE) tissue arrays, which contained 80 cases (Cat. #LC818c), and normal lung tissue arrays, which contained 24 cases (Cat. #LCN241), were purchased from US Biomax.

Techniques: Expressing, Formalin-fixed Paraffin-Embedded, Immunohistochemistry, Staining, Software

Journal: Molecular cancer therapeutics

Article Title: Protein phosphatase 2A as a therapeutic target in small cell lung cancer

doi: 10.1158/1535-7163.MCT-21-0013

Figure Lengend Snippet: (A) Scatter plot shows an upregulation of the PP2A-A subunit in the tumor samples (p=0.0144). A Mann-Whitney U test was used for comparison between the normal and SCLC samples. (B) IHC for PP2A was conducted on TMA tissue sections and images were captured at 4x or 20x using a 3D-Histech PANNORAMIC SCAN whole slide scanner (3D-Histech, Budapest, Hungary). PP2A subunit A positively immunostained the cytoplasm and nucleus of normal lung and tumor tissue but was highly upregulated in tumor tissue. TMAs were scored in normal (n=24) and tumor (n=79) cores on a scale from 0 (no staining/no protein expression) to 3+ (strong staining/high protein expression). (C) Summary bar graph of the average PP2A subunit staining. IHC staining intensity of normal and tumor cores. There was a statistically significant difference between normal and tumor tissue (***, p<0.001). Student’s t test was used for comparison between the normal and SCLC samples. (D) In order to compare the expression of PP2A subunits A and C, cell lysates from seven SCLC cell lines and HBEC 3KT (non-malignant cell line) were subjected to western blotting (n=3 biological replicates). (E) PP2A activity was determined using a serine/threonine phosphatase activity assay (Millipore) after 24 h exposure to cantharidin (10 µM) and LB100 (5 µM) (n=3 biological replicates). ***, p<0.001, results were analyzed by ANOVA with Tukey post-test. (F) The inset showed reduction of PP2A subunit Aα in H524 cells as well as inhibition of cell proliferation due to PP2A subunit Aα knockdown (n=3 biological replicates). p<0.05, Student’s t test was used for comparison between the groups. LB100 alone or in combination with carboplatin inhibited proliferation and colony formation in SCLC cells. The Cell Counting Kit-8 assay detected cell H524 and H69 cell viability. (n=3 biological replicates). (G, H) Cells were treated with LB100, carboplatin and etoposide, as a single treatment or in combination, at constant ratio. The combination index (CI) was calculated using Chou-Talalay method to find synergism between LB100 with carboplatin and etoposide (CompuSyn software: www.combosyn.com). **, p<0.01, ANOVA with Tukey post-test was used for comparison between LB100, LB100/carboplatin and LB100/etoposide. Colony formation assays were used to count the ability of H524 (I) and H69 (J) cells to form colonies. Drug concentrations are listed for two assays with H524 and H69 respectively: LB100 (2.5 µM; 20 µM), carboplatin (4 µM; 20 µM), etoposide (3 µM; 30 µM), LB100/carboplatin (2.5&4 µM; 20&20 µM) and LB100/etoposide (2.5&3 µM; 20&30 µM). Representative images of colonies at 4x are shown under the graph (n=2). *p<0.05; **, p<0.01; ***, p<0.001; ****, p<0.0001. Results were analyzed by ANOVA with Tukey post-test.

Article Snippet: Tissue Microarray Small cell lung cancer TMAs were from US Biomax Inc. (Rockville, MD; LC818).

Techniques: MANN-WHITNEY, Comparison, Staining, Expressing, Immunohistochemistry, Western Blot, Activity Assay, Phosphatase Assay, Inhibition, Knockdown, Cell Counting, Software

Journal: Molecular cancer therapeutics

Article Title: Protein phosphatase 2A as a therapeutic target in small cell lung cancer

doi: 10.1158/1535-7163.MCT-21-0013

Figure Lengend Snippet: (A) Significant changes were observed for signal transduction and metabolic pathways. (B) MicroArray analysis showed that overnight treatment with 20 µM treatment with LB100 inhibited utilization of carbon substrate sources. Table includes 10 carbon sources affected by LB100 (n=3). (C) LB100 significantly inhibited two carbon substrates utilization by H69 cells. P < 0.001 (***) for control (untreated cells) vs. LB100. Results were analyzed by ANOVA with Tukey post-test. (D) Amplex Red Glucose/Oxidase assay kit was used to measure glucose level in cell culture media. Glucose level was significantly higher in cell culture medium from cells treated with LB100 (20 µM). Glucose concentration detected in initial medium and counted as 100%. Subtracting final medium level of glucose from initial glucose medium concentration yielded % glucose in the medium with cells. Level of glucose dropped in control with cells and in LB100 treated groups (p < 0.0001 (****). Results were analyzed by ANOVA with Tukey post-test, (n=3 biological replicates). Effect of LB100 on MET phosphorylation. (E) H524 and H69 cells were treated overnight with LB100 (H524 – 5 µM and H69 – 20 µM) following by stimulation with 100 ng/ml HGF in 10 min. Cells were collected and lysed for WB analysis with pMET and total MET antibody. Pan-actin was used as loading control (n=3 biological replicates). (F) H524 cell lysates (control, LB100, carboplatin and combination (LB100/carboplatin) were analyzed by western blots to check phosphorylation status of MET at Ser985 and Tyr1234/1235. Actin was used as a loading control (n=3 biological replicates).

Article Snippet: Tissue Microarray Small cell lung cancer TMAs were from US Biomax Inc. (Rockville, MD; LC818).

Techniques: Transduction, Microarray, Control, Glucose Oxidase Assay, Cell Culture, Concentration Assay, Western Blot