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-{"Images": [{"Name": "i0", "Description": "Cell State Dependent Effects of Bmal1 on Melanoma Immunity and Tumorigenicity", "Path": "https://s3.amazonaws.com/www.cycif.org/zang-2023", "Width": 70784, "Height": 50041, "MaxLevel": 7}], "Header": "The circadian clock regulator Bmal1 modulates tumorigenesis, but its reported effects are inconsistent. Here, we show that Bmal1 has a context-dependent role in mouse melanoma tumor growth. Loss of Bmal1 in YUMM2.1 or B16-F10 melanoma cells eliminated clock function and diminished hypoxic gene expression and tumorigenesis, which could be rescued by ectopic expression of HIF-1a in YUMM2.1 cells. By contrast, over-expressed wild-type or a transcriptionally inactive mutant Bmal1 non-canonically sequestered myosin heavy chain 9 (Myh9) to increase MRTF-SRF activity and AP-1 transcriptional signature, and shift YUMM 2.1 cells from a Sox10high to a Sox9high immune resistant, mesenchymal cell state that is found in human melanomas. Our work uncovers a link between Bmal1, Myh9, mouse melanoma cell plasticity, and tumor immunity. This connection may underlie cancer therapeutic resistance and underpin the link between the circadian clock, MRTF-SRF and the cytoskeleton. ", "Rotation": 0, "Layout": {"Grid": [["i0"]]}, "Stories": [{"Name": "", "Description": "", "Waypoints": [{"Name": "Lineage switch in dHLH-Bmal1", "Description": "Over-expressed wildtype or a transcriptionally inactive mutant Bmal1 shifts YUMM 2.1 cells from a Sox10-high to a Sox9-high immune-resistant, mesenchymal cell state that is found in human melanomas.", "Arrows": [{"Text": "YUMM2.1 EV", "HideArrow": true, "Point": [0.2643563704059749, 0.056446707562739284], "Angle": 60}, {"Text": "YUMM2.1 WT-Bmal1", "HideArrow": true, "Point": [0.7333383402450123, 0.05031171576714627], "Angle": 60}, {"Text": "YUMM2.1 dHLH-Bmal1", "HideArrow": true, "Point": [1.1964905417608742, 0.05448717286981852], "Angle": 60}], "Overlays": [], "Group": "Proliferation", "Masks": [], "ActiveMasks": [], "Zoom": 1.7196694214876034, "Pan": [1.1185984143839258, 0.36396991378914045]}, {"Name": "Ki67 in SOX10-high cells", "Description": "YUMM2.1-WT tumors grew faster than the more anti-PD1 resistant YUMM2.1-dHLH tumors. Correlating with slower tumor growth rates of YUMM2.1-dHLH tumors, the percentage of Ki67+ melanoma cells (Sox9+ or Sox10+ cells) was lower in YUMM2.1-dHLH compared to YUMM2.1-WT tumors", "Arrows": [{"Text": "YUMM2.1 EV", "HideArrow": true, "Point": [0.2643563704059749, 0.056446707562739284], "Angle": 60}, {"Text": "YUMM2.1 WT-Bmal1", "HideArrow": true, "Point": [0.7333383402450123, 0.05031171576714627], "Angle": 60}, {"Text": "YUMM2.1 dHLH-Bmal1", "HideArrow": true, "Point": [1.1964905417608742, 0.05448717286981852], "Angle": 60}], "Overlays": [], "Group": "Lineage markers", "Masks": [], "ActiveMasks": [], "Zoom": 0.35256198347107437, "Pan": [0.7072600467615198, 0.118722075830167]}]}], "Channels": [{"Rendered": false, "Name": "DNA", "Path": "DNA_0__DNA"}, {"Rendered": false, "Name": "SOX9", "Path": "SOX9_1__SOX9"}, {"Rendered": false, "Name": "SOX10", "Path": "SOX10_2__SOX10"}, {"Rendered": false, "Name": "Ki67", "Path": "Ki67_3__Ki67"}], "Masks": [], "Groups": [{"Name": "Proliferation", "Colors": ["0000ff", "ffff00", "ff00ff", "00ffff"], "Channels": ["DNA", "SOX9", "SOX10", "Ki67"], "Descriptions": ["", "", "", ""]}, {"Name": "Lineage markers", "Colors": ["0000ff", "ffff00", "ff00ff"], "Channels": ["DNA", "SOX9", "SOX10"], "Descriptions": ["", "", ""]}], "FirstViewport": {"Zoom": 0.5, "Pan": [0.6, 0.4]}, "FirstArrows": [{"Text": "YUMM2.1 EV", "HideArrow": true, "Point": [0.2643563704059749, 0.056446707562739284], "Angle": 60}, {"Text": "YUMM2.1 WT-Bmal1", "HideArrow": true, "Point": [0.7333383402450123, 0.05031171576714627], "Angle": 60}, {"Text": "YUMM2.1 dHLH-Bmal1", "HideArrow": true, "Point": [1.1964905417608742, 0.05448717286981852], "Angle": 60}]}
+{"Images": [{"Name": "i0", "Description": "Cell State Dependent Effects of Bmal1 on Melanoma Immunity and Tumorigenicity", "Path": "https://s3.amazonaws.com/www.cycif.org/zang-2023", "Width": 70784, "Height": 50041, "MaxLevel": 7}], "Header": "Here we show\u00a0six tissue cores from mouse melanoma tissues. The cores represent\u00a0two replicate mice (columns)\u00a0across three mouse lines (rows):  \n1. wild-type Bmal1 (YUMM2.1-EV)  \n2. a transcriptionally inactive Bmal1 mutant lacking the basic helix-loop-helix DNA binding domain (dHLH) (YUMM2.1-dHLH-Bmal1)  \n3. empty vector (YUMM2.1-EV)  \n\nUsing multiplexed\u00a0imaging,\u00a0we found\u00a0an\u00a0increase in Sox9 and decrease in Sox10 expression in WT-Bmal1\u00a0tumors\u00a0and in dHLH-Bmal1 expressing tumors,\u00a0indicating a shift in cell lineage.  \n\n### Attribution  \n**Please cite the publication and underlying data as:**  \nZhang et al.,\u00a0Cell State Dependent Effects of Bmal1 on Melanoma Immunity and Tumorigenicity,\u00a02023,\u00a0_Nature Communications_. [DOI](www.doi/org).  \n\n**Please cite this Minerva story as:**  \nShishir M Pant, Cecily C. Ritch,\u00a0Juliann Tefft, John Hoffer. Cell State Dependent Effects of Bmal1 on Melanoma Immunity and Tumorigenicity, 2023.  \n\n### Experimental Details  \n**Mouse background:** C57BL/6  \n**Mouse Tumor Line:** YUMM2.1  \n\n## Imaging  \n**Assay:** CyCIF  \n**Fixative Type:** Formalin  \n**Microscope:** CyteFinder II HT  \n**Objective:** 20X/0.75 NA  \n\n### Associated Identifiers  \n**LSP slide ID:** LSP12583\n\n", "Rotation": 0, "Layout": {"Grid": [["i0"]]}, "Stories": [{"Name": "", "Description": "", "Waypoints": [{"Name": "Lineage switch in dHLH-Bmal1", "Description": "Over-expressed wildtype or a transcriptionally inactive mutant Bmal1 shifts YUMM 2.1 cells from a Sox10-high to a Sox9-high immune-resistant, mesenchymal cell state that is found in human melanomas.", "Arrows": [], "Overlays": [], "Group": "Lineage markers", "Masks": [], "ActiveMasks": [], "Zoom": 1.7026092089728453, "Pan": [1.1185984143839258, 0.36396991378914045]}, {"Name": "Ki67 in SOX10-high cells", "Description": "YUMM2.1-WT tumors grew faster than the YUMM2.1-dHLH tumors, which are more anti-PD1 resistant. \n\nThe percentage of Ki67+ melanoma cells (Sox9+ or Sox10+ cells) was lower in YUMM2.1-dHLH compared to YUMM2.1-WT tumors, which correlated with the slower growth rates of the YUMM2.1-dHLH tumors. ", "Arrows": [{"Text": "YUMM2.1 EV", "HideArrow": true, "Point": [0.2643563704059749, 0.056446707562739284], "Angle": 60}, {"Text": "YUMM2.1 WT-Bmal1", "HideArrow": true, "Point": [0.7333383402450123, 0.05031171576714627], "Angle": 60}, {"Text": "YUMM2.1 dHLH-Bmal1", "HideArrow": true, "Point": [1.1964905417608742, 0.05448717286981852], "Angle": 60}], "Overlays": [], "Group": "Proliferation", "Masks": [], "ActiveMasks": [], "Zoom": 8.544596262911998, "Pan": [0.2218668783813088, 0.6362344064066255]}]}], "Channels": [{"Rendered": false, "Name": "DNA", "Path": "DNA_0__DNA"}, {"Rendered": false, "Name": "SOX9", "Path": "SOX9_1__SOX9"}, {"Rendered": false, "Name": "SOX10", "Path": "SOX10_2__SOX10"}, {"Rendered": false, "Name": "Ki67", "Path": "Ki67_3__Ki67"}], "Groups": [{"Name": "Proliferation", "Colors": ["0000ff", "ffff00", "ff00ff", "00ffff"], "Channels": ["DNA", "SOX9", "SOX10", "Ki67"], "Descriptions": ["", "", "", ""]}, {"Name": "Lineage markers", "Colors": ["0000ff", "ffff00", "ff00ff"], "Channels": ["DNA", "SOX9", "SOX10"], "Descriptions": ["", "", ""]}], "Masks": [], "PixelsPerMicron": 3.0769230769230766}