Journal: JCI Insight

Article Title: Collectin-11 promotes cancer cell proliferation and tumor growth

doi: 10.1172/jci.insight.159452

Figure Lengend Snippet: Based on our findings and literature interpretation, we propose that CL-11 in the tumor (locally synthesized and circulation derived) can contribute to melanoma tumor growth via 2 pathways: (a) CL-11–induced activation of mitogenic kinase signaling and stimulation of tumor cell proliferation, possibly through interaction with ErbB receptors (e.g., EGFR, HER2) on melanoma cells, and(b) CL-11–dependent promotion of immunosuppressive TME, characterized by a high proportion of myeloid cells and a low proportion of lymphocytes, lack of cytotoxic T cell infiltrating in the tumor core, increased angiogenesis, low levels of intratumor expression of cytokines/chemokines with antitumor properties (e.g., Ifng, Nos2, Il12, Ccl5, Cxcl9 ), and MΦ reprogramming to a M2 phenotype.

Article Snippet: The following antibodies were used in signaling pathway studies: anti–phospho-ERK1/2 (Thr202/Tyr204, 197G2), anti–phospho-Akt (Ser473, D9E), anti–phospho-JNK (Thr183/Tyr185, 81E11), anti–phospho-HER1 (Tyr1068, D7A5), anti–phospho-HER2 (Tyr1221/1222, 6B12), anti–phospho-HER3 (Tyr1289, D1B5), and anti-ERK1/2, anti-Akt, anti-JNK, anti-HER1(D38B1), anti-HER2 (D8F12), anti-HER3 (D22C5), and anti-HER4 (111B2) (all from Cell Signaling Technology).

Techniques: Synthesized, Derivative Assay, Activation Assay, Expressing

Journal: Cancer Cell

Article Title: A living biobank of patient-derived ductal carcinoma in situ mouse-intraductal xenografts identifies risk factors for invasive progression

doi: 10.1016/j.ccell.2023.04.002

Figure Lengend Snippet: A living biobank of MIND-PDX models of DCIS retaining histological and molecular features of primary lesions (A) Schematic overview of the generation and characterization of 130 MIND-PDX models of DCIS. (B) Pie chart of the grade distribution of the primary DCIS lesions. (C) Pie chart of the DCIS take rate in MIND-PDX models. (D) Examples of H&E-stained sections of the different growth patterns observed in primary DCIS lesions (top row) or DCIS-MIND outgrowths (middle row). Bottom row: human-specific Ku80 staining showing that DCIS-MIND outgrowths have a human origin. (E) Distribution of growth patterns of primary DCIS lesions and the corresponding DCIS-MIND outgrowths. A black line indicates concordance between primary and PDX, whereas an orange line indicates a discordance between primary and PDX. (F) Growth pattern analyses between an early (3–6 months) and late (12 months) time points. (G) Examples of immunohistochemistry for ER, PR, HER2, and Ki67 expression in DCIS-MIND lesions (top row) vs. matched primary DCIS lesions (bottom row). (H) Distribution of molecular subtypes (luminal A: ER + , PR +/− , HER2 − , Ki67 < 20%; luminal B: ER + , PR +/− , HER2 – , Ki67 ≥ 20%; HER2 + : ER +/− , PR +/− , and HER2 + ; basal: ER − , PR − , HER2 − ) of primary DCIS lesions and the corresponding DCIS-MIND lesions. A black line indicates concordance between primary and PDX, whereas an orange line indicates a discordance between primary and PDX. See also Figure S1 and Table S1 .

Article Snippet: Phospho-HER2/ErbB2 Antibody (Clone Tyr1248) , Cell Signaling , Cat# 2247; RRID: AB_331725.

Techniques: Staining, Immunohistochemistry, Expressing

Journal: Cancer Cell

Article Title: A living biobank of patient-derived ductal carcinoma in situ mouse-intraductal xenografts identifies risk factors for invasive progression

doi: 10.1016/j.ccell.2023.04.002

Figure Lengend Snippet: DCIS-MIND models retain mutational and transcriptional features of the primary DCIS lesions (A) Oncoprint showing the mutational landscape of the primary DCIS lesions, including amplifications, single-nucleotide variants, and insertion-deletions (indels) for the top mutated genes in our breast cancer gene panel. Annotations for each model includes ER, PR, and HER2 status. (B) Oncoprint showing amplifications, single-nucleotide variants, and insertion-deletions (indels) in cancer genes in primary DCIS lesions and corresponding DCIS-MIND lesions for the top mutated genes. (C) Unsupervised clustering of DCIS-MIND lesions based on PAM50 genes, showing clustering of luminal, HER2 + , and basal-like DCIS lesions. Annotations include origin (primary or PDX) and molecular subtype based on PAM50 or IHC. (D) Unsupervised clustering of DCIS-MIND lesions based on 90 informative genes resulting in three DCIS subtypes proposed by Strand et al. See also Figure S2 and Tables S2 and .

Article Snippet: Phospho-HER2/ErbB2 Antibody (Clone Tyr1248) , Cell Signaling , Cat# 2247; RRID: AB_331725.

Techniques:

Journal: Cancer Cell

Article Title: A living biobank of patient-derived ductal carcinoma in situ mouse-intraductal xenografts identifies risk factors for invasive progression

doi: 10.1016/j.ccell.2023.04.002

Figure Lengend Snippet: Comparison of clinical biomarkers related to invasive progression of DCIS (A) Whole-mount analysis (top row), H&E staining (middle row), and human-specific Ku80 staining (bottom row) of DCIS-MIND lesions showing non-invasive growth (left panel) or invasive growth (right panel). Cells of human origin are marked with Ku80 (green), myoepithelial cells are marked with alpha-smooth muscle actin (αSMA) (magenta). Blue arrows/circles indicate invasive cells. (B) Pie chart of the percentage of DCIS-MIND lesions with non-invasive growth, micro-invasion, or invasive progression. (C) Odds ratio table showing risk scores (univariate and multivariate linear regression models) for association between common clinical parameters, DCIS subtypes as proposed by Strand et al., and multiple invasive recurrence classifiers for invasive progression of DCIS-MIND lesions, identifying HER2 overexpression and solid growth patterns as independent risk factors. Molecular subtype and growth pattern parameters are based on DCIS-MIND characteristics while the other parameters are based on patient characteristics. See also Figure S3 .

Article Snippet: Phospho-HER2/ErbB2 Antibody (Clone Tyr1248) , Cell Signaling , Cat# 2247; RRID: AB_331725.

Techniques: Staining, Over Expression

Journal: Cancer Cell

Article Title: A living biobank of patient-derived ductal carcinoma in situ mouse-intraductal xenografts identifies risk factors for invasive progression

doi: 10.1016/j.ccell.2023.04.002

Figure Lengend Snippet: HER2 overexpression is a driver for invasive progression of DCIS (A) Representative IHC images of p-ERK and p-AKT. (B) Plotted H scores for p-ERK and p-AKT expression in HER2 + DCIS and HER2 – DCIS controls. (C) Tumor-free survival curves of responsive HER2 + DCIS models treated with vehicle (C) or herceptin (T) with log rank test. ∗∗∗p < 0.001, ∗∗p < 0.01, ∗p < 0.05. (D) Tumor-free survival curves of non-responsive HER2 + DCIS models treated with vehicle (C) or herceptin (T) with log rank test. ∗∗∗p < 0.001, ∗∗p < 0.01, ∗p < 0.05. (E) Representative whole-mount images and Ku80-stained sections of DCIS-injected mammary glands from DCIS063 mice treated with vehicle (left panels) or herceptin (right panels). (F) Schematic representation of the experimental setup for lentiviral overexpression of HER2 in HER2 – DCIS cells. (G) Western blot showing expression of HER2 and phospho-HER2 in parental 293T cells and 293T cells transduced with the HER2-GFP lentivirus. (H) Representative whole-mount images of intraductally injected mammary glands, showing non-invasive replacement growth of non-transduced HER2 – DCIS cells (left panel) and invasive expansive growth of the same DCIS cells transduced with the HER2-GFP lentivirus (right panel). Inset shows lesion without αSMA marker indicating GFP expression (green). Cells of human origin are marked with Ku80 (gray), myoepithelial cells are marked with αSMA (magenta). Inset shows lesion without αSMA marker indicating HER2-GFP expression (green). See also Figure S7 .

Article Snippet: Phospho-HER2/ErbB2 Antibody (Clone Tyr1248) , Cell Signaling , Cat# 2247; RRID: AB_331725.

Techniques: Over Expression, Expressing, Staining, Injection, Western Blot, Transduction, Marker

Journal: Cancer Cell

Article Title: A living biobank of patient-derived ductal carcinoma in situ mouse-intraductal xenografts identifies risk factors for invasive progression

doi: 10.1016/j.ccell.2023.04.002

Figure Lengend Snippet:

Article Snippet: Phospho-HER2/ErbB2 Antibody (Clone Tyr1248) , Cell Signaling , Cat# 2247; RRID: AB_331725.

Techniques: Labeling, Recombinant, Electron Microscopy, Plasmid Preparation, Blocking Assay, Software

Journal: Cancer Cell

Article Title: A living biobank of patient-derived ductal carcinoma in situ mouse-intraductal xenografts identifies risk factors for invasive progression

doi: 10.1016/j.ccell.2023.04.002

Figure Lengend Snippet: A living biobank of MIND-PDX models of DCIS retaining histological and molecular features of primary lesions (A) Schematic overview of the generation and characterization of 130 MIND-PDX models of DCIS. (B) Pie chart of the grade distribution of the primary DCIS lesions. (C) Pie chart of the DCIS take rate in MIND-PDX models. (D) Examples of H&E-stained sections of the different growth patterns observed in primary DCIS lesions (top row) or DCIS-MIND outgrowths (middle row). Bottom row: human-specific Ku80 staining showing that DCIS-MIND outgrowths have a human origin. (E) Distribution of growth patterns of primary DCIS lesions and the corresponding DCIS-MIND outgrowths. A black line indicates concordance between primary and PDX, whereas an orange line indicates a discordance between primary and PDX. (F) Growth pattern analyses between an early (3–6 months) and late (12 months) time points. (G) Examples of immunohistochemistry for ER, PR, HER2, and Ki67 expression in DCIS-MIND lesions (top row) vs. matched primary DCIS lesions (bottom row). (H) Distribution of molecular subtypes (luminal A: ER + , PR +/− , HER2 − , Ki67 < 20%; luminal B: ER + , PR +/− , HER2 – , Ki67 ≥ 20%; HER2 + : ER +/− , PR +/− , and HER2 + ; basal: ER − , PR − , HER2 − ) of primary DCIS lesions and the corresponding DCIS-MIND lesions. A black line indicates concordance between primary and PDX, whereas an orange line indicates a discordance between primary and PDX. See also Figure S1 and Table S1 .

Article Snippet: HER2/ErbB2 Rabbit mAb (Clone 29D8) , Cell Signaling , Cat# 2165; RRID: AB_10692490.

Techniques: Staining, Immunohistochemistry, Expressing

Journal: Cancer Cell

Article Title: A living biobank of patient-derived ductal carcinoma in situ mouse-intraductal xenografts identifies risk factors for invasive progression

doi: 10.1016/j.ccell.2023.04.002

Figure Lengend Snippet: DCIS-MIND models retain mutational and transcriptional features of the primary DCIS lesions (A) Oncoprint showing the mutational landscape of the primary DCIS lesions, including amplifications, single-nucleotide variants, and insertion-deletions (indels) for the top mutated genes in our breast cancer gene panel. Annotations for each model includes ER, PR, and HER2 status. (B) Oncoprint showing amplifications, single-nucleotide variants, and insertion-deletions (indels) in cancer genes in primary DCIS lesions and corresponding DCIS-MIND lesions for the top mutated genes. (C) Unsupervised clustering of DCIS-MIND lesions based on PAM50 genes, showing clustering of luminal, HER2 + , and basal-like DCIS lesions. Annotations include origin (primary or PDX) and molecular subtype based on PAM50 or IHC. (D) Unsupervised clustering of DCIS-MIND lesions based on 90 informative genes resulting in three DCIS subtypes proposed by Strand et al. See also Figure S2 and Tables S2 and .

Article Snippet: HER2/ErbB2 Rabbit mAb (Clone 29D8) , Cell Signaling , Cat# 2165; RRID: AB_10692490.

Techniques:

Journal: Cancer Cell

Article Title: A living biobank of patient-derived ductal carcinoma in situ mouse-intraductal xenografts identifies risk factors for invasive progression

doi: 10.1016/j.ccell.2023.04.002

Figure Lengend Snippet: Comparison of clinical biomarkers related to invasive progression of DCIS (A) Whole-mount analysis (top row), H&E staining (middle row), and human-specific Ku80 staining (bottom row) of DCIS-MIND lesions showing non-invasive growth (left panel) or invasive growth (right panel). Cells of human origin are marked with Ku80 (green), myoepithelial cells are marked with alpha-smooth muscle actin (αSMA) (magenta). Blue arrows/circles indicate invasive cells. (B) Pie chart of the percentage of DCIS-MIND lesions with non-invasive growth, micro-invasion, or invasive progression. (C) Odds ratio table showing risk scores (univariate and multivariate linear regression models) for association between common clinical parameters, DCIS subtypes as proposed by Strand et al., and multiple invasive recurrence classifiers for invasive progression of DCIS-MIND lesions, identifying HER2 overexpression and solid growth patterns as independent risk factors. Molecular subtype and growth pattern parameters are based on DCIS-MIND characteristics while the other parameters are based on patient characteristics. See also Figure S3 .

Article Snippet: HER2/ErbB2 Rabbit mAb (Clone 29D8) , Cell Signaling , Cat# 2165; RRID: AB_10692490.

Techniques: Staining, Over Expression

Journal: Cancer Cell

Article Title: A living biobank of patient-derived ductal carcinoma in situ mouse-intraductal xenografts identifies risk factors for invasive progression

doi: 10.1016/j.ccell.2023.04.002

Figure Lengend Snippet: HER2 overexpression is a driver for invasive progression of DCIS (A) Representative IHC images of p-ERK and p-AKT. (B) Plotted H scores for p-ERK and p-AKT expression in HER2 + DCIS and HER2 – DCIS controls. (C) Tumor-free survival curves of responsive HER2 + DCIS models treated with vehicle (C) or herceptin (T) with log rank test. ∗∗∗p < 0.001, ∗∗p < 0.01, ∗p < 0.05. (D) Tumor-free survival curves of non-responsive HER2 + DCIS models treated with vehicle (C) or herceptin (T) with log rank test. ∗∗∗p < 0.001, ∗∗p < 0.01, ∗p < 0.05. (E) Representative whole-mount images and Ku80-stained sections of DCIS-injected mammary glands from DCIS063 mice treated with vehicle (left panels) or herceptin (right panels). (F) Schematic representation of the experimental setup for lentiviral overexpression of HER2 in HER2 – DCIS cells. (G) Western blot showing expression of HER2 and phospho-HER2 in parental 293T cells and 293T cells transduced with the HER2-GFP lentivirus. (H) Representative whole-mount images of intraductally injected mammary glands, showing non-invasive replacement growth of non-transduced HER2 – DCIS cells (left panel) and invasive expansive growth of the same DCIS cells transduced with the HER2-GFP lentivirus (right panel). Inset shows lesion without αSMA marker indicating GFP expression (green). Cells of human origin are marked with Ku80 (gray), myoepithelial cells are marked with αSMA (magenta). Inset shows lesion without αSMA marker indicating HER2-GFP expression (green). See also Figure S7 .

Article Snippet: HER2/ErbB2 Rabbit mAb (Clone 29D8) , Cell Signaling , Cat# 2165; RRID: AB_10692490.

Techniques: Over Expression, Expressing, Staining, Injection, Western Blot, Transduction, Marker

Journal: Cancer Cell

Article Title: A living biobank of patient-derived ductal carcinoma in situ mouse-intraductal xenografts identifies risk factors for invasive progression

doi: 10.1016/j.ccell.2023.04.002

Figure Lengend Snippet:

Article Snippet: HER2/ErbB2 Rabbit mAb (Clone 29D8) , Cell Signaling , Cat# 2165; RRID: AB_10692490.

Techniques: Labeling, Recombinant, Electron Microscopy, Plasmid Preparation, Blocking Assay, Software

Journal: Communications Biology

Article Title: CRISPR-clear imaging of melanin-rich B16-derived solid tumors

doi: 10.1038/s42003-023-04614-7

Figure Lengend Snippet: a Comparison of antibody labeling of intact melanin (+/–) tumors. HER2 is detected with an anti-HER2 antibody and stained with an Alexa-488 labeled secondary antibody (shown in white), tdTomato-expressing B16 cells (shown in purple), imaged at increasing depths. Images were acquired using a 25x objective. Scale bar 150 μm (yellow). b Overview of workflow for 3D imaging, localization and visualization of nanoparticles in melanin(–) tumors. The method consists of 2 modular steps: (1) multi-dye vessel-staining (highlighted in Fig. ) and injection of nanoparticles in tissue, and (2) clearing for high imaging quality using 3D confocal microscopy. For clarity, separate channels are shown for nanoparticles (Alexa 488), tdTomato-expressing melanin(–) B16 melanoma cells and vasculature (Alexa 647). Scale bar 300 μm. A 10x/0.45 objective was used for imaging. c 3D visualization of digital reconstruction of nanoparticles at 500 µm depth of melanin(–) tumors cleared with PACT . Purple is vasculature, cyan are reconstructed tdTomato-expressing melanin(–) B16 melanoma cells, and green represents nanoparticles. Scale bar 300 μm. White box insert on the left is a blow-up of the box on the right. Scale bar 20 μm for insert. A 10x/0.45 objective was used for imaging.

Article Snippet: Expression of HER2 and tdTomato was tested in experiments without α-MSH stimulation, with the following primary antibodies (1:1000 dilution): Anti-HER2/ErbB2 antibody (Cell Signaling, #2165), anti-tdTomato antibody (KeraFAST, EST203), using goat anti-rabbit IgG (H + L) secondary antibody-HRP (Thermo Fisher Scientific, #31460, and HRP goat anti-rat IgG antibody (Vector Laboratories, PI-9400).

Techniques: Antibody Labeling, Staining, Labeling, Expressing, Imaging, Injection, Confocal Microscopy

Journal: Investigational New Drugs

Article Title: Anticancer effect of zanubrutinib in HER2-positive breast cancer cell lines

doi: 10.1007/s10637-023-01346-7

Figure Lengend Snippet: Anticancer effects of BTK inhibitors in breast cancer cell lines

Article Snippet: The following specific antibodies were used and purchased from Cell Signaling: anti-EGFR (D38B1), anti-HER2/ErbB2 (D8F12), anti-HER3/ErbB3 (D22C5), anti-HER4/ErbB4 (111B2), anti-phospho-EGFR Y1068 (D7A5), anti-phospho-HER2/ErbB2 Y1221/1222 (6B12), anti-phospho-HER3/ErbB3 Y1289 (D1B5), anti-phospho-HER4/ErbB4 Y1284/EGFR Y1173 (21A9), anti-Akt (pan) (C67E7), anti-phospho-Akt S473 (D9E), anti-p44/42 MAPK (Erk1/2), anti-phospho-p44/42 MAPK (phospho-Erk1/2) T202/Y204), and anti-PARP (46D11).

Techniques:

Journal: Investigational New Drugs

Article Title: Anticancer effect of zanubrutinib in HER2-positive breast cancer cell lines

doi: 10.1007/s10637-023-01346-7

Figure Lengend Snippet: Effects of BTK inhibitors on signalling pathways in HER2-positive breast cancer cell lines. Compounds were used at a 10 µM concentration for 16 h of treatment. Cells were stimulated by heregulin (HRG, 0.1 µg/mL) 30 min prior to harvesting. β-Actin served as a control for equal loading. The representative results from at least two biological replicates are shown

Article Snippet: The following specific antibodies were used and purchased from Cell Signaling: anti-EGFR (D38B1), anti-HER2/ErbB2 (D8F12), anti-HER3/ErbB3 (D22C5), anti-HER4/ErbB4 (111B2), anti-phospho-EGFR Y1068 (D7A5), anti-phospho-HER2/ErbB2 Y1221/1222 (6B12), anti-phospho-HER3/ErbB3 Y1289 (D1B5), anti-phospho-HER4/ErbB4 Y1284/EGFR Y1173 (21A9), anti-Akt (pan) (C67E7), anti-phospho-Akt S473 (D9E), anti-p44/42 MAPK (Erk1/2), anti-phospho-p44/42 MAPK (phospho-Erk1/2) T202/Y204), and anti-PARP (46D11).

Techniques: Concentration Assay

Journal: Investigational New Drugs

Article Title: Anticancer effect of zanubrutinib in HER2-positive breast cancer cell lines

doi: 10.1007/s10637-023-01346-7

Figure Lengend Snippet: Dose-dependent effect of zanubrutinib in HER2-positive BT474 and SKBR3 cell lines. Cells were treated with increasing concentrations of zanubrutinib for 16 h. Cells were stimulated by heregulin (HRG, 0.1 µg/mL) 30 min prior to harvesting. β-Actin served as a control for equal loading. The representative results from at least two biological replicates are shown

Article Snippet: The following specific antibodies were used and purchased from Cell Signaling: anti-EGFR (D38B1), anti-HER2/ErbB2 (D8F12), anti-HER3/ErbB3 (D22C5), anti-HER4/ErbB4 (111B2), anti-phospho-EGFR Y1068 (D7A5), anti-phospho-HER2/ErbB2 Y1221/1222 (6B12), anti-phospho-HER3/ErbB3 Y1289 (D1B5), anti-phospho-HER4/ErbB4 Y1284/EGFR Y1173 (21A9), anti-Akt (pan) (C67E7), anti-phospho-Akt S473 (D9E), anti-p44/42 MAPK (Erk1/2), anti-phospho-p44/42 MAPK (phospho-Erk1/2) T202/Y204), and anti-PARP (46D11).

Techniques:

Journal: Investigational New Drugs

Article Title: Anticancer effect of zanubrutinib in HER2-positive breast cancer cell lines

doi: 10.1007/s10637-023-01346-7

Figure Lengend Snippet: Heregulin rescues HER2-positive cells from G1 arrest induced by BTK inhibitors. Cells were treated for 24 h with increasing concentrations of the selected compounds, with or without activation by heregulin (HRG, 0.1 µg/ml). Results are averages of biological triplicates, the error bars represent standard deviation

Article Snippet: The following specific antibodies were used and purchased from Cell Signaling: anti-EGFR (D38B1), anti-HER2/ErbB2 (D8F12), anti-HER3/ErbB3 (D22C5), anti-HER4/ErbB4 (111B2), anti-phospho-EGFR Y1068 (D7A5), anti-phospho-HER2/ErbB2 Y1221/1222 (6B12), anti-phospho-HER3/ErbB3 Y1289 (D1B5), anti-phospho-HER4/ErbB4 Y1284/EGFR Y1173 (21A9), anti-Akt (pan) (C67E7), anti-phospho-Akt S473 (D9E), anti-p44/42 MAPK (Erk1/2), anti-phospho-p44/42 MAPK (phospho-Erk1/2) T202/Y204), and anti-PARP (46D11).

Techniques: Activation Assay, Standard Deviation

Journal: Investigational New Drugs

Article Title: Anticancer effect of zanubrutinib in HER2-positive breast cancer cell lines

doi: 10.1007/s10637-023-01346-7

Figure Lengend Snippet: (A) Ibrutinib and zanubrutinib inhibit colony formation in HER2 + SKBR3 cells, while the proliferation rate of HER2- MCF7 cells remains unaffected. Colonies were stained with crystal violet after 10 days of treatment. The numbers indicate the percentages of colonies formed (calculated from the absorbance values, numbers are averages of biological duplicates ± SD). (B) Immunoblot analysis of lysates of cells treated with 1 µM zanubrutinib and ibrutinib. β-Actin served as a control for equal loading. The representative images are shown

Article Snippet: The following specific antibodies were used and purchased from Cell Signaling: anti-EGFR (D38B1), anti-HER2/ErbB2 (D8F12), anti-HER3/ErbB3 (D22C5), anti-HER4/ErbB4 (111B2), anti-phospho-EGFR Y1068 (D7A5), anti-phospho-HER2/ErbB2 Y1221/1222 (6B12), anti-phospho-HER3/ErbB3 Y1289 (D1B5), anti-phospho-HER4/ErbB4 Y1284/EGFR Y1173 (21A9), anti-Akt (pan) (C67E7), anti-phospho-Akt S473 (D9E), anti-p44/42 MAPK (Erk1/2), anti-phospho-p44/42 MAPK (phospho-Erk1/2) T202/Y204), and anti-PARP (46D11).

Techniques: Staining, Western Blot

Journal: JCI Insight

Article Title: Collectin-11 promotes cancer cell proliferation and tumor growth

doi: 10.1172/jci.insight.159452

Figure Lengend Snippet: Based on our findings and literature interpretation, we propose that CL-11 in the tumor (locally synthesized and circulation derived) can contribute to melanoma tumor growth via 2 pathways: (a) CL-11–induced activation of mitogenic kinase signaling and stimulation of tumor cell proliferation, possibly through interaction with ErbB receptors (e.g., EGFR, HER2) on melanoma cells, and(b) CL-11–dependent promotion of immunosuppressive TME, characterized by a high proportion of myeloid cells and a low proportion of lymphocytes, lack of cytotoxic T cell infiltrating in the tumor core, increased angiogenesis, low levels of intratumor expression of cytokines/chemokines with antitumor properties (e.g., Ifng, Nos2, Il12, Ccl5, Cxcl9 ), and MΦ reprogramming to a M2 phenotype.

Article Snippet: The following antibodies were used in signaling pathway studies: anti–phospho-ERK1/2 (Thr202/Tyr204, 197G2), anti–phospho-Akt (Ser473, D9E), anti–phospho-JNK (Thr183/Tyr185, 81E11), anti–phospho-HER1 (Tyr1068, D7A5), anti–phospho-HER2 (Tyr1221/1222, 6B12), anti–phospho-HER3 (Tyr1289, D1B5), and anti-ERK1/2, anti-Akt, anti-JNK, anti-HER1(D38B1), anti-HER2 (D8F12), anti-HER3 (D22C5), and anti-HER4 (111B2) (all from Cell Signaling Technology).

Techniques: Synthesized, Derivative Assay, Activation Assay, Expressing