Fig. 2: Mesenchymal neuroblastomas show characteristics of senescence. | Cell Death & Disease

Fig. 2: Mesenchymal neuroblastomas show characteristics of senescence.

From: High content-imaging drug synergy screening identifies specific senescence-related vulnerabilities of mesenchymal neuroblastomas

Fig. 2

A Left: Comparison of phenotypic features measured with Cell Profiler-based image analysis of cells stained with Hoechst (Nuclei), CellMask (Cell membrane), Lysotracker (functional lysosomes) and LAMP1 (Lysosomal membrane protein) of mesenchymal and adrenergic cells. Feature measurements are the mean of six replicates. Right: Exemplary images of the mesenchymal (Gi-M-EN) and adrenergic (IMR-32) phenotype. Scale bar 100 µm. Nuclei are colored in blue, Cell membranes in red and lysosomes in green. B Segmentation based on cellular and lysosomal staining and feature extraction from segmented objects in the high-content imaging pipeline. Scale bar 50 µm C ssGSEA of senescence secretory phenotype-associated pathways in correlation with microscopically determined lysoscore (green color). calculated from the sum of three lysosomal measurements normalized to the Gi-M-EN cell line. Gene sets: SIR: Pribluda_SIR.gense. SASP_Pri: Pribluda_SASP.genes. Biocarta_Inflammation. GO_Inflammation. Biocarta_nfkb. OIS_Acosta. OIS_Kuilman. SASP_Coppe2008. DDSR_Rodier. SMS_Kuilman. WP_SPHINGOLIPID_METABOLISM_IN_SENESCENCE. D Pearson’s correlation between mes/adr score and lysosomal score of 24 neuroblastoma cell lines. E Uniform manifold approximation and projection (UMAP) embedding of single cell data of the cell line SK-N-SH published by Jansky et al. 2021. Neuroblastoma subtypes are annotated on the left plot based on Jansky et al. 2021. Expression of SASP-associated genes is shown in the middle plot; TFE3 expression is shown on the right. F Pearson’s correlation of mes/adr score and gene log2 expression of LAMP2 and TFE3. G Correlation of the lysosomal score and the DSSasym for MEK inhibitors. H Correlation of the MPAS score and the lysosomal score. I Left: Schematic description of the workflow for processing, preculturing and drug sensitivity profiling of patient-derived fresh tissue samples obtained through the INFORM program. Right: Heatmap representing the ssGSEA scores for gene sets linked to senescence-associated secretome signaling in five INFORM fresh tissue patient samples, DSSasym scores for three MEKi in the bottom annotation.

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