+{"Images": [{"Name": "i0", "Description": "MYCN-Amplified Ependymoma ", "Path": "https://s3.amazonaws.com/www.cycif.org/gonzalez-castro-2026/p211-ependymoma", "Width": 30506, "Height": 24438, "MaxLevel": 5}], "Header": "This sample is from a 55-year-old female who presented with progressive radiating lower back pain. An MRI of the spine was obtained demonstrating two homogeneously enhancing intradural and extramedullary lesions, one at C3 and the other spanning the T12 to L2 spinal levels. Pathologic evaluation revealed a densely cellular glial neoplasm with brisk mitotic activity, microvascular proliferation, perivascular pseudorosettes, and focal necrosis, histologically consistent with ependymoma. Molecular profiling revealed MYCN amplification and variants of uncertain significance (VUS) in PDGFRA (c.1786+7T>C) and APC (c.5399A>G). The overall features were consistent with a diagnosis of Spinal ependymoma, MYCN-amplified, CNS W.H.O. grade 3. \n\nThe patient subsequently experienced intracranial dissemination following standard therapy. HER2 immunostaining was performed on specimen from the patient\u2019s initial operation, motivated by recent systematic profiling of antibody-drug conjugate (ADC) targets in central nervous system tumors [1], showing 2+ to 3+ staining . Multiplexed immunofluorescence imaging confirmed HER2 expression across both quiescent tumor regions and highly proliferative zones. Notably, we found that while HER2 is broadly expressed, EGFR and Ki-67 co-localize to highly proliferative compartments and are inversely correlated with CD44 expression.\n\n[1] Coy et al., Neuro Oncol. 2024 Mar 4;26(3):458-472. \n\n \n\n### **Metadata about this sample**\n### Clinical Context:\nMultiply recurrent MYCN-amplified spinal ependymoma with intracranial dissemination, treated with HER2-directed ADC following failure of standard therapies. \n \n### Diagnosis \n**Primary Diagnosis:** Ependymoma \n**WHO Classification:** WHO grade 2 ependymoma \n**Molecular subtype:** MYCN-amplified spinal ependymoma \n**Anatomic Site(s):** Cervical (C3) and thoracolumbar spine (T12\u2013L2) \n**Age at Diagnosis:** 55 years \n**Tumor Grade:** WHO grade 3 \n**Stage at Diagnosis:** Not applicable (primary CNS tumor) \n\n### Demographics \n**Species:** Homo sapiens \n**Sex:** Female \n**Vital Status:** Deceased\n\n### Therapy \n**Initial Disease Status:** Primary diagnosis \n**Pre-operative Therapy:** None \n**Post-operative Therapy:** \n - Radiotherapy \n - Chemotherapy \n - HER2-directed antibody\u2013drug conjugate (trastuzumab deruxtecan) \n\n### Follow-Up\n**Progression or Recurrence:** Yes \n**Last Known Disease Status:** Deceased (medical complications including pneumonia and pulmonary embolism) \n**Age at Last Follow-Up:** ~57 years \n**Days to Progression:** Not specified \n\n### Biospecimen \n**Acquisition Method:** Surgical resection \n**Specimen Type:** FFPE tissue section \n\n### Molecular Testing \n**Molecular Analysis Method:** Targeted next-generation sequencing \n**Genotype Available:** Yes \n**Key Genetic Features:** MYCN amplification \n\n### Imaging \n**Imaging Assay Type:** CyCIF \n**Fixative:** Formalin \n**Microscope:** RareCyte CyteFinder \n**Objective:** 20\u00d7 / 0.75 NA \n**Pixel Binning:** Yes \n\n### **Publication and Data Availability** \n\n### Associated Publication: \n\nClinical responses to trastuzumab deruxtecan in molecularly defined ependymoma (Submitted, 2026)\n\n### Attribution: \n\n**Please cite the publication and underlying data as:** \nGonzalez Castro LN, Wu Y-C, Lin J-R, Santiago-Ribeiro M-J, Laurenge A, Carpentier A, Coy S, Lin NU, Mokhtari K, Ligon KL, Touat M, Santagata S.\nMYCN-amplified spinal ependymoma with HER2 expression (CyCIF).\nHarvard Dataverse, 2026. DOI: XXXXX \n\n**Please cite this Minerva Story as:** \nGonzalez Castro LN, Wu Y-C, Lin J-R, Santiago-Ribeiro M-J, Laurenge A, Carpentier A, Coy S, Lin NU, Mokhtari K, Ligon KL, Touat M, Santagata S. MYCN-Amplified Ependymoma. 2026, http://127.0.0.1:2020/\n\n### Associated Identifiers\n| ID Type | ID | \n| **LSP Slide ID** | LSP33564 | \n| **Patient LSP ID** | LSP34837 | \n| **Block LSP ID** | LSP34839 | \n| **Section LSP ID** | LSP62883 |\n", "Rotation": 0, "Layout": {"Grid": [["i0"]]}, "Stories": [{"Name": "", "Description": "", "Waypoints": [{"Name": "HER2 [low power]", "Description": "At low power, HER2 showed strong and diffuse expression, with some focal intra-tumoral heterogeneity. ", "Arrows": [], "Overlays": [], "Group": "HER expression", "Masks": [], "ActiveMasks": [], "Zoom": 0.5124375000000001, "Pan": [0.5323996420379489, 0.5018396283404194]}, {"Name": "HER2 [high power]", "Description": "A higher power view demonstrated strong and diffuse membranous HER2 expression, with apparent circumferential expression on tumor cells. ", "Arrows": [], "Overlays": [], "Group": "HER expression", "Masks": [], "ActiveMasks": [], "Zoom": 19.106650856155806, "Pan": [1.0598520498567126, 0.3258180048606602]}, {"Name": "EGFR-positive proliferative nodules", "Description": "Multiplexed imaging showed that while HER2 was broadly expressed across much of the tumor landscape, EGFR and Ki-67 demonstrated focal expression in discrete tumor nodules, indicating that they exhibited a distinct proliferative phenotype. In contrast, CD44 was enriched in adjacent non-nodular regions, demonstrating an inverse spatial relationship with both EGFR and Ki-67.", "Arrows": [], "Overlays": [], "Group": "Overview", "Masks": [], "ActiveMasks": [], "Zoom": 3.7195798982428863, "Pan": [1.0601042607240088, 0.30466946990293853]}, {"Name": "Nodule proliferation", "Description": "There was a strong correlation between EGFR and Ki67 labeling in proliferative areas, and these regions showed low CD44 expression. ", "Arrows": [], "Overlays": [], "Group": "Proliferation", "Masks": [], "ActiveMasks": [], "Zoom": 10.96988859362404, "Pan": [1.0983206556619831, 0.29917580928774234]}, {"Name": "Signaling", "Description": "Examination of intracellular signaling proteins via multiplexed imaging showed distinctive patterning of effectors such as pERK, indicative of focal activation of the MAPK signaling pathway. ", "Arrows": [], "Overlays": [], "Group": "Signaling pathway", "Masks": [], "ActiveMasks": [], "Zoom": 3.645091982738807, "Pan": [1.02209441358314, 0.30924586925578024]}, {"Name": "Epigenetics", "Description": "Multiplexed imaging showed patterning of epigenetic markers, with variable levels of H3K27me3, H3K27ac, and H3K4me3 in proliferative EGFR-positive tumor nodules. ", "Arrows": [], "Overlays": [], "Group": "Epigenetic markers ", "Masks": [], "ActiveMasks": [], "Zoom": 5.248541317823894, "Pan": [1.0605242604586747, 0.3135018012450722]}, {"Name": "Cilia", "Description": "Upon closer examination, we observed the presence of cilia on ependymoma tumor cells. Staining with gTUB highlighted the basal body, marking the site where the cilium originates, while ARL13b staining clearly visualized the ciliary structures themselves (arrows). Numerous primary and multiciliated cells were observed in proliferative nodules and adjacent CD44-positive regions. ", "Arrows": [{"Text": "", "HideArrow": false, "Point": [1.0906310743614525, 0.28289110803082845], "Angle": 60}, {"Text": "", "HideArrow": false, "Point": [1.0920688040042263, 0.28175761349667994], "Angle": 60}, {"Text": "", "HideArrow": false, "Point": [1.0938718975470694, 0.2829455339484354], "Angle": 60}, {"Text": "", "HideArrow": false, "Point": [1.0951729532799444, 0.2832071592860244], "Angle": 60}, {"Text": "", "HideArrow": false, "Point": [1.0896936270697564, 0.2825881687514808], "Angle": 60}], "Overlays": [], "Group": "Cillia", "Masks": [], "ActiveMasks": [], "Zoom": 88.38975694444447, "Pan": [1.0903712602639621, 0.2827737813220676]}, {"Name": "Additional Nodules", "Description": "Numerous proliferative nodules were observed in this patient sample. A separate region of the tumor was shown, again demonstrating strong expression of EGFR and Ki-67 in nodular regions, with CD44 predominantly expressed in adjacent regions. ", "Arrows": [{"Text": "", "HideArrow": false, "Point": [1.1083672276773968, 0.5429861233799362], "Angle": 60}, {"Text": "", "HideArrow": false, "Point": [0.9809225258223961, 0.5558757006105767], "Angle": 60}, {"Text": "", "HideArrow": false, "Point": [0.9053240371020215, 0.5022419823319342], "Angle": 60}], "Overlays": [], "Group": "Overview", "Masks": [], "ActiveMasks": [], "Zoom": 1.341228261931011, "Pan": [0.9255169707748778, 0.43098422600560443]}]}], "Channels": [{"Rendered": false, "Name": "Hoechst_1", "Path": "Hoechst-1_0__Hoechst-1"}, {"Rendered": false, "Name": "Ki67", "Path": "Ki67_9__Ki67"}, {"Rendered": false, "Name": "HER2", "Path": "HER2_11__HER2"}, {"Rendered": false, "Name": "CD44", "Path": "CD44_20__CD44"}, {"Rendered": false, "Name": "CD31", "Path": "CD31_22__CD31"}, {"Rendered": false, "Name": "EGFR", "Path": "EGFR_24__EGFR"}, {"Rendered": false, "Name": "PCNA", "Path": "PCNA_39__PCNA"}, {"Rendered": false, "Name": "SOX2", "Path": "SOX2_45__SOX2"}, {"Rendered": false, "Name": "SMA", "Path": "SMA_17__SMA"}, {"Rendered": false, "Name": "FOXJ1", "Path": "FOXJ1_6__FOXJ1"}, {"Rendered": false, "Name": "gTUB", "Path": "gTUB_14__gTUB"}, {"Rendered": false, "Name": "H2ax", "Path": "H2ax_10__H2ax"}, {"Rendered": false, "Name": "pERK", "Path": "pERK_29__pERK"}, {"Rendered": false, "Name": "LaminB", "Path": "LaminB_27__LaminB"}, {"Rendered": false, "Name": "LaminA", "Path": "LaminA_44__LaminA"}, {"Rendered": false, "Name": "PDPN", "Path": "PDPN_26__PDPN"}, {"Rendered": false, "Name": "S100", "Path": "S100_19__S100"}, {"Rendered": false, "Name": "ARL13b", "Path": "ARL13b_7__ARL13b"}, {"Rendered": false, "Name": "H3K4me3", "Path": "H3K4me3_46__H3K4me3"}, {"Rendered": false, "Name": "H3K27me3", "Path": "H3K27me3_25__H3K27me3"}, {"Rendered": false, "Name": "H3K27ac", "Path": "H3K27ac_41__H3K27ac"}], "Masks": [], "Groups": [{"Name": "Overview", "Colors": ["737171", "f7f4f4", "ff0000", "0000ff", "00ffff", "00ff00"], "Channels": ["Hoechst_1", "Ki67", "HER2", "CD44", "CD31", "EGFR"], "Descriptions": ["", "", "", "", "", ""]}, {"Name": "All", "Colors": ["737171", "ff0000", "00ff00", "0000ff", "f7f4f4", "ff7800", "6703f6", "f1a073", "be8cf1", "00ffff", "12d393", "4b84ed", "e782cd", "fc06be", "fa8b08", "00d7ff", "e80ae8", "f1210b", "ff0000", "00ff00", "002eff"], "Channels": ["Hoechst_1", "HER2", "EGFR", "CD44", "Ki67", "PCNA", "SOX2", "CD31", "SMA", "FOXJ1", "gTUB", "H2ax", "pERK", "LaminB", "LaminA", "PDPN", "S100", "ARL13b", "H3K4me3", "H3K27me3", "H3K27ac"], "Descriptions": ["", "", "", "", "", "", "", "", "", "", "", "", "", "", "", "", "", "", "", "", ""]}, {"Name": "Proliferation", "Colors": ["737171", "00ffff", "ff0000", "00ff00", "0025ff"], "Channels": ["Hoechst_1", "PCNA", "Ki67", "EGFR", "CD44"], "Descriptions": ["", "", "", "", ""]}, {"Name": "Tumor", "Colors": ["777777", "ff0000", "00ffff", "00ff00", "0000ff"], "Channels": ["Hoechst_1", "HER2", "SOX2", "EGFR", "CD44"], "Descriptions": ["", "", "", "", ""]}, {"Name": "Nuclear morphology", "Colors": ["737171", "00ff00", "7b09f5"], "Channels": ["Hoechst_1", "LaminB", "LaminA"], "Descriptions": ["", "", ""]}, {"Name": "Tissue structure", "Colors": ["737171", "00ff25", "ffffff", "09e6f2", "ff0000", "1000ff"], "Channels": ["Hoechst_1", "PDPN", "CD31", "SMA", "S100", "FOXJ1"], "Descriptions": ["", "", "", "", "", ""]}, {"Name": "Epigenetic markers ", "Colors": ["737171", "00ff00", "002eff", "ff0000"], "Channels": ["Hoechst_1", "H3K27me3", "H3K27ac", "H3K4me3"], "Descriptions": ["", "", "", ""]}, {"Name": "Signaling pathway", "Colors": ["737171", "ff0000", "ffffff", "00ff00", "0001ff"], "Channels": ["Hoechst_1", "EGFR", "pERK", "CD44", "FOXJ1"], "Descriptions": ["", "", "", "", ""]}, {"Name": "HER expression", "Colors": ["737171", "00ff00"], "Channels": ["Hoechst_1", "HER2"], "Descriptions": ["", ""]}, {"Name": "Cillia", "Colors": ["737171", "ffffff", "ff0000", "00ff00"], "Channels": ["Hoechst_1", "FOXJ1", "gTUB", "ARL13b"], "Descriptions": ["", "", "", ""]}, {"Name": "sample info", "Colors": ["777777", "ff0000", "0000ff", "00ff00", "ff00ff", "00ffff"], "Channels": ["Hoechst_1", "HER2", "CD44", "EGFR", "Ki67", "CD31"], "Descriptions": ["", "", "", "", "", ""]}], "PixelsPerMicron": 1.5384615948919735, "FirstGroup": "sample info", "FirstViewport": {"Zoom": 0.42703125000000014, "Pan": [0.4972925451346908, 0.485822054262618]}}
0 commit comments