Journal: Thoracic Cancer

Article Title: Tumor suppressor effect of an antibody on xenotransplanted sarcomatoid mesothelioma cells

doi: 10.1111/1759-7714.14591

Figure Lengend Snippet: Representative immunofluorescent staining of cultured mesothelioma cells using AX10 antibody (a), immunohistochemical staining (b), and secondary antibody‐drug conjugate assay in vitro (c). (a) AX10 immunoreactivity in MPM‐1, −2, and −3 cells, representing sarcomatoid, epithelioid, and biphasic type mesothelioma, respectively. All MPM‐1, −2, and −3 cells exhibited AX10 antibody immunoreactivity at the cell surface. The staining was analyzed using a Guava easyCyte cell analyzer and accompanying software to obtain a one‐parameter log histogram. (b) AX10 immunoreactivity in various mesothelioma tissue specimens. Weak or no AX10 immunoreactivity was detected in five out of 10 epithelioid mesothelioma tissues (a). One out of five biphasic mesotheliomas exhibited AX10 immunoreactivity in spindle sarcomatoid components (arrow) but weak immunoreactivity in epithelioid components (arrowhead) (b). Five out of six sarcomatoid mesothelioma tissues exhibited strong AX10 immunoreactivity (c). Little AX10 immunoreactivity was detected in normal human tissues. No significant AX10 immunoreactivity was detected in the lung (d) (pleural mesothelial cells; insert) tissue specimens. Weak AX10 immunoreactivity was detected in myofibrous cells in the uterus (e). We did not detect any significant AX10 immunoreactivity in the brain, liver, or kidney, whereas strong AX10 immunoreactivity was observed in a nonmelanocytic (hypomelanocytic) melanoma tissue sample that was supplementally included in the microarray (f) (staining without AX10 antibody; insert). (c) MPM‐1 sarcomatoid mesothelioma cells were incubated with AX10 at 10, 100, and 1000 ng/mL followed by incubation with anti‐murine IgG (Fc) antibody conjugated to duocarmycin. Representative staining with Annexin V‐PI is presented. Note the dose‐dependent Annexin V‐positive and PI‐negative apoptotic MPM‐1 cells in the presence of AX10 antibody

Article Snippet: Tissue microarrays composed of mesothelioma (Cat. No. MS801b) and Food and Drug Administration (FDA) normal organ tissue arrays (Cat. No. NBP2‐78057) were purchased from US Biomax and Novus Biologicals, respectively.

Techniques: Staining, Cell Culture, Immunohistochemical staining, In Vitro, Software, Microarray, Incubation

Journal: Thoracic Cancer

Article Title: Tumor suppressor effect of an antibody on xenotransplanted sarcomatoid mesothelioma cells

doi: 10.1111/1759-7714.14591

Figure Lengend Snippet: AX10 does not affect cell proliferation, but significantly decreases Matrigel invasion activity of MPM‐1 sarcomatoid mesothelioma cells in vitro. (a) Representative cell proliferation assay. At 24 h, the cell number was 1.80 ± 0.10 (mock) and 1.77 ± 0.06 (AX10). Respective numbers at 48 h were 2.40 ± 0.10 (mock) and 2.37 ± 0.12 (AX10), while at 72 h they were 3.90 ± 0.20 (mock) and 4.20 ± 0.61 (AX10). The data represent means ± SD from triplicate assays (Student's t ‐test, p > 0.5). (b) AX10 significantly reduced Matrigel invasion activity of MPM‐1 cells (Student's t ‐test, p < 0.01). The number of invading cells was 59.7 ± 7.02 (mock) and 10.3 ± 1.52 (AX10) at 24 h, and 210.7 ± 11.4 (mock) and 15.0 ± 3.00 (AX10) at 48 h. Data from triplicate assays are expressed as means ± SD ( n = 3). (c) Cells that migrated to the lower surface of the membrane are shown (48 h). Original magnification, ×100

Article Snippet: Tissue microarrays composed of mesothelioma (Cat. No. MS801b) and Food and Drug Administration (FDA) normal organ tissue arrays (Cat. No. NBP2‐78057) were purchased from US Biomax and Novus Biologicals, respectively.

Techniques: Activity Assay, In Vitro, Proliferation Assay, Membrane

Journal: Thoracic Cancer

Article Title: Tumor suppressor effect of an antibody on xenotransplanted sarcomatoid mesothelioma cells

doi: 10.1111/1759-7714.14591

Figure Lengend Snippet: Inhibitory effect of AX10 on MPM‐1 xenotransplanted sarcomatoid mesothelioma cell proliferation. (a) Inoculation of AX10 antibody delayed the growth of xenotransplanted MPM‐1 sarcomatoid mesothelioma tumors. On day 0, SCID‐NOD mice were subcutaneously implanted with MPM‐1 cells. The following day, day 3, the mice were administered AX10 antibody or vehicle only by intraperitoneal injection and weekly thereafter as indicated by arrows. Values are represented as means ± standard error for n = 5 mice. Statistical significance was measured by a two‐sided unpaired Student's t ‐test (* p < 0.01). (b) On day 42, the xenotransplanted tumors were excised to determine their weight. Total tumor weights are represented as means ± standard error for n = 5 mice. Statistical significance was measured by a two‐sided unpaired Student's t ‐test ( p < 0.01). (c) Gross and histological appearance of a representative xenotransplanted tumor. Arrowhead indicates the tumor without AX10 antibody, while the arrow indicates the small tumor remaining following weekly AX10 injection. Note the elimination of tumor cells, which were histologically replaced by regenerative muscle in mice inoculated with AX10 antibody. Scale bar indicates 100 μm

Article Snippet: Tissue microarrays composed of mesothelioma (Cat. No. MS801b) and Food and Drug Administration (FDA) normal organ tissue arrays (Cat. No. NBP2‐78057) were purchased from US Biomax and Novus Biologicals, respectively.

Techniques: Injection

Journal: Cancer Research

Article Title: Integrating AI-Powered Digital Pathology and Imaging Mass Cytometry Identifies Key Classifiers of Tumor Cells, Stroma, and Immune Cells in Non–Small Cell Lung Cancer

doi: 10.1158/0008-5472.CAN-23-1698

Figure Lengend Snippet: Multidimensional analysis of the NSCLC tumor ecosystem by IMC. A, Schematic representation of the IMC workflow on a formalin-fixed, paraffin-embedded tissue microarray. Key steps include staining with metal-tagged antibodies, spot-by-spot laser ablation, and acquisition by a mass cytometer. High dimensional images are reconstructed, processed, and segmented at both cellular and tissue level, generating data for further analyses. B, Heat map showing the mean values of key lineage markers adopted for cell cluster annotation. Proteins and cell phenotypes are ordered by hierarchical clustering with the Pearson correlation distance. Protein expression is color-coded from blue (lower) to red (higher) and scaled by column. C, Representative matched pictures of a NSCLC specimen showing pan-cytokeratin–positive tumor cells (left) and the tissue segmentation resulting from the machine learning pixel classifier (right). D, Spatial distribution and quantification of immune cell populations as the absolute cell number per mm 2 (left) or as a percentage of total immune cells (right) in the tumor and the stroma. E, Heat map showing the normalized marker expression in each macrophage cluster. Markers and cell clusters are ordered by hierarchical clustering according to Pearson correlation distance. Mean values of marker expression are represented and color-coded from blue (lower) to red (higher) and scaled by column. Color code indicates cluster identity. F and G, UMAP projections of macrophage cells ( n = 46733) from NSCLC tumors showing 20 clusters ( F ) or the cell distribution according to tissue segmentation ( G ). Each dot represents an individual cell. H, S100A8 + Mϕ infiltrate both the stroma and the tumor nests of NSCLC tissues. Representative pictures of the distribution of Mϕ (defined as CD68 + cells) and the subpopulation of S100A8 + Mϕ within tumor nests of a NSCLC tissue.

Article Snippet: Cohort 1 included 108 patients diagnosed with NSCLC, whose tumor specimen was spotted on a tissue microarray (TMA; LC2162–US Biomax), containing 209 cores.

Techniques: Formalin-fixed Paraffin-Embedded, Microarray, Staining, Cytometry, Expressing, Marker

Journal: EBioMedicine

Article Title: Loss of SLC25A11 causes suppression of NSCLC and melanoma tumor formation

doi: 10.1016/j.ebiom.2019.01.036

Figure Lengend Snippet: Expression and distribution of SLC25A11 in lung cancer and malignant melanoma tissue samples. (a) Tissue microarray was performed to determine the expression of SLC25A11 in normal lung tissues and lung cancer samples. A-1: Normal lung tissue showing no staining (×200; scale bar, 200 μm); A-2: Weak, A-3: Moderate, and A-4: Strong expression of SLC25A11 in lung cancer samples (×400; scale bar, 60 μm). (b) Percent distribution (graph) and SLC25A11 expression patterns in normal and cancer samples (Table) according to histologic subtypes. Statistical significance compared with normal lung tissues was determined by Fisher's exact test. (c) The expression patterns of SLC25A11 in malignant melanoma were determined by IHC. A-1: Weak, A-2: Moderate, A-3: Strong (×400; scale bar, 60 μm). (d) Percent SLC25A11 expression in normal skin and malignant melanoma samples. (Statistical significance was calculated by Fisher's Exact test. ***p < .001, **p < .01, *p < .05).

Article Snippet: Tissue microarray (TMA) from formalin-fixed, paraffin-embedded tissue blocks of NSCLC including normal lung tissue (CCN, CC, Super Bio Chips, Seoul, Korea) and malignant melanoma patients were purchased (ME2082b melanoma TMA: US Biomax, Bethesda, MD, USA).

Techniques: Expressing, Microarray, Staining