ORCID Profile
0000-0001-7464-6187
Current Organisation
Garvan Institute of Medical Research
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Publisher: American Association for Cancer Research (AACR)
Date: 11-08-2023
DOI: 10.1158/2767-9764.23932418.V1
Abstract: Supplementary Video 1B shows representative images of a naltrindole-treated spheroid. Scale bar: 200 µm, number at top left: days post-implantation.
Publisher: American Association for Cancer Research (AACR)
Date: 17-07-2023
DOI: 10.1158/2767-9764.23696146
Abstract: Supplementary Table 1 shows Cell type isolated, age (days) and number of cells sorted for each spheroid included in the study
Publisher: American Association for Cancer Research (AACR)
Date: 17-07-2023
DOI: 10.1158/2767-9764.23696152.V1
Abstract: Supplementary Figure 8 shows a summary of GSEA pathway analyses using Hallmarks gene set showing top 25 enriched pathways for each comparison
Publisher: American Association for Cancer Research (AACR)
Date: 11-08-2023
DOI: 10.1158/2767-9764.CRC-22-0305
Abstract: Solid cancer cells escape the primary tumor mass by transitioning from an epithelial-like state to an invasive migratory state. As they escape, metastatic cancer cells employ interchangeable modes of invasion, transitioning between fibroblast-like mesenchymal movement to amoeboid migration, where cells display a rounded morphology and navigate the extracellular matrix in a protease-independent manner. However, the gene transcripts that orchestrate the switch between epithelial, mesenchymal, and amoeboid states remain incompletely mapped, mainly due to a lack of methodologies that allow the direct comparison of the transcriptomes of spontaneously invasive cancer cells in distinct migratory states. Here, we report a novel single-cell isolation technique that provides detailed three-dimensional data on melanoma growth and invasion, and enables the isolation of live, spontaneously invasive cancer cells with distinct morphologies and invasion parameters. Via the expression of a photoconvertible fluorescent protein, compact epithelial-like cells at the periphery of a melanoma mass, elongated cells in the process of leaving the mass, and rounded amoeboid cells invading away from the mass were tagged, isolated, and subjected to single-cell RNA sequencing. A total of 462 differentially expressed genes were identified, from which two candidate proteins were selected for further pharmacologic perturbation, yielding striking effects on tumor escape and invasion, in line with the predictions from the transcriptomics data. This work describes a novel, adaptable, and readily implementable method for the analysis of the earliest phases of tumor escape and metastasis, and its application to the identification of genes underpinning the invasiveness of malignant melanoma. This work describes a readily implementable method that allows for the isolation of in idual live tumor cells of interest for downstream analyses, and provides the single-cell transcriptomes of melanoma cells at distinct invasive states, both of which open avenues for in-depth investigations into the transcriptional regulation of the earliest phases of metastasis.
Publisher: American Association for Cancer Research (AACR)
Date: 17-07-2023
DOI: 10.1158/2767-9764.23696149
Abstract: Supplementary Figure 9 shows Escaped cell analyses for naltrindole and sulfasalazine spheroid assays
Publisher: American Association for Cancer Research (AACR)
Date: 11-08-2023
DOI: 10.1158/2767-9764.23932466.V1
Abstract: Supplementary Figure 1 shows data generation for the photoconvertible spheroid assay
Publisher: American Association for Cancer Research (AACR)
Date: 11-08-2023
DOI: 10.1158/2767-9764.23932472.V1
Abstract: Inhibition of OPRD1 reduces melanoma spheroid growth and invasion. b A, /b Representative maximum intensity projections of imaged hemispheroids for vehicle- and naltrindole-treated spheroids, scale bar 200 μm. b B, /b Relative spheroid volume over time for vehicle- and naltrindole-treated cells. Shaded areas represent SEM. b C, /b Cumulative number of escaped cells over time normalized to spheroid surface area. B and C, Datapoints represent the mean of eight replicates.
Publisher: American Association for Cancer Research (AACR)
Date: 11-08-2023
DOI: 10.1158/2767-9764.23932424.V1
Abstract: Supplementary Table 6 shows Sorting events for each cell included in the final data set
Publisher: American Association for Cancer Research (AACR)
Date: 17-07-2023
DOI: 10.1158/2767-9764.23696140
Abstract: Supplementary Table 3 shows Differentially expressed genes for escaping vs. epithelial analysis
Publisher: American Association for Cancer Research (AACR)
Date: 17-07-2023
DOI: 10.1158/2767-9764.23696143
Abstract: Supplementary Table 2 shows Transcript counts for common housekeeping genes, with cells excluded from the study due to low housekeeping expression highlighted in yellow
Publisher: American Association for Cancer Research (AACR)
Date: 11-08-2023
DOI: 10.1158/2767-9764.23932460
Abstract: Supplementary Figure 3 shows validation of OPRD1 and SLC19A1 protein expression
Publisher: American Association for Cancer Research (AACR)
Date: 11-08-2023
DOI: 10.1158/2767-9764.23932451
Abstract: Supplementary Figure 6 shows an Amoeboid cell cohort gallery
Publisher: American Association for Cancer Research (AACR)
Date: 11-08-2023
DOI: 10.1158/2767-9764.23932430.V1
Abstract: Supplementary Table 4 shows Differentially expressed genes for amoeboid vs. escaping analysis
Publisher: American Association for Cancer Research (AACR)
Date: 11-08-2023
DOI: 10.1158/2767-9764.23932454
Abstract: Supplementary Figure 5 shows an Escaping cell cohort gallery
Publisher: American Association for Cancer Research (AACR)
Date: 11-08-2023
DOI: 10.1158/2767-9764.23932427.V1
Abstract: Supplementary Table 5 shows Differentially expressed genes for epithelial vs. amoeboid analysis
Publisher: American Association for Cancer Research (AACR)
Date: 11-08-2023
DOI: 10.1158/2767-9764.23932412
Abstract: Supplementary Video 2B shows representative images of a Sulfasalazine-treated spheroid. Scale bar: 200 µm, number at top left: days post-implantation.
Publisher: American Association for Cancer Research (AACR)
Date: 11-08-2023
DOI: 10.1158/2767-9764.23932457.V1
Abstract: Supplementary Figure 4 shows an Epithelial cohort gallery
Publisher: American Association for Cancer Research (AACR)
Date: 11-08-2023
DOI: 10.1158/2767-9764.23932457
Abstract: Supplementary Figure 4 shows an Epithelial cohort gallery
Publisher: American Association for Cancer Research (AACR)
Date: 11-08-2023
DOI: 10.1158/2767-9764.23932415
Abstract: Supplementary Video 2A shows representative images of a vehicle-treated spheroid. Scale bar: 200 µm, number at top left: days post-implantation.
Publisher: American Association for Cancer Research (AACR)
Date: 11-08-2023
DOI: 10.1158/2767-9764.23932415.V1
Abstract: Supplementary Video 2A shows representative images of a vehicle-treated spheroid. Scale bar: 200 µm, number at top left: days post-implantation.
Publisher: American Association for Cancer Research (AACR)
Date: 17-07-2023
DOI: 10.1158/2767-9764.23696143.V1
Abstract: Supplementary Table 2 shows Transcript counts for common housekeeping genes, with cells excluded from the study due to low housekeeping expression highlighted in yellow
Publisher: American Association for Cancer Research (AACR)
Date: 11-08-2023
DOI: 10.1158/2767-9764.23932418
Abstract: Supplementary Video 1B shows representative images of a naltrindole-treated spheroid. Scale bar: 200 µm, number at top left: days post-implantation.
Publisher: American Association for Cancer Research (AACR)
Date: 11-08-2023
DOI: 10.1158/2767-9764.23932469.V1
Abstract: SSZ treatment slows spheroid growth but induces tumor escape by elongated cells. b A, /b Representative maximum intensity projections of imaged hemispheroids for vehicle- and SSZ-treated spheroids. Scale bar, 200 μm. b B, /b Relative spheroid volume for vehicle- and SSZ-treated spheroids. b C, /b Cumulative escaped cells per surface area of imaged hemispheroid. b D, /b Mean escaped cell sphericity per day. B–D, Shaded areas represent SEM.
Publisher: American Association for Cancer Research (AACR)
Date: 17-07-2023
DOI: 10.1158/2767-9764.23696158
Abstract: Supplementary Figure 6 shows an Amoeboid cell cohort gallery
Publisher: American Association for Cancer Research (AACR)
Date: 17-07-2023
DOI: 10.1158/2767-9764.23696128.V1
Abstract: Supplementary Video 1A shows representative daily images of a vehicle-treated spheroid. Scale bar: 200 µm, number at top left: days post-implantation.
Publisher: American Association for Cancer Research (AACR)
Date: 11-08-2023
DOI: 10.1158/2767-9764.23932448.V1
Abstract: Supplementary Figure 7 shows metadata for isolated cells
Publisher: American Association for Cancer Research (AACR)
Date: 17-07-2023
DOI: 10.1158/2767-9764.23696149.V1
Abstract: Supplementary Figure 9 shows Escaped cell analyses for naltrindole and sulfasalazine spheroid assays
Publisher: American Association for Cancer Research (AACR)
Date: 17-07-2023
DOI: 10.1158/2767-9764.23696119
Abstract: Supplementary Video 2B shows representative images of a Sulfasalazine-treated spheroid. Scale bar: 200 µm, number at top left: days post-implantation.
Publisher: American Association for Cancer Research (AACR)
Date: 17-07-2023
DOI: 10.1158/2767-9764.23696170.V1
Abstract: Supplementary Figure 2 shows clustering of 44 single cells and their gene expression heat map based on 230 differentially expressed genes
Publisher: The Company of Biologists
Date: 03-2020
DOI: 10.1242/JCS.238014
Abstract: It has become increasingly evident that T cell functions are subject to translational control in addition to transcriptional regulation. Here, by using live imaging of CD8+ T cells isolated from the Lifeact-EGFP mouse, we show that T cells exhibit a gain in fluorescence intensity following engagement of cognate tumour target cells. The GFP signal increase is governed by Erk1/2-dependent distal T cell receptor (TCR) signalling and its magnitude correlates with IFN-γ and TNF-α production, which are hallmarks of T cell activation. Enhanced fluorescence was due to increased translation of Lifeact-EGFP protein, without an associated increase in its mRNA. Activation-induced gains in fluorescence were also observed in naïve and CD4+ T cells from the Lifeact-EGFP reporter, and were readily detected by both flow cytometry and live cell microscopy. This unique, translationally controlled reporter of effector T cell activation simultaneously enables tracking of cell morphology, F-actin dynamics and activation state in in idual migrating T cells. It is a valuable addition to the limited number of reporters of T cell dynamics and activation, and opens the door to studies of translational activity and heterogeneities in functional T cell responses in situ.
Publisher: American Association for Cancer Research (AACR)
Date: 11-08-2023
DOI: 10.1158/2767-9764.23932475.V1
Abstract: Gene expression analysis for epithelial, escaping, and amoeboid cells. b A, /b Top: 462 genes were DE, 144 for escaping versus epithelial (Esc-v-Epi), 22 for amoeboid versus escaping (Amo-v-Esc), and 422 for amoeboid versus epithelial (Amo-v-Epi). Bottom: Venn diagram of the number of DE genes for the three pairwise comparisons. b B, /b Volcano plot for Esc-v-Epi DE analysis, with genes enriched in “escaping” cells shown as positive on the i x /i -axis. b C, /b Volcano plot for Amo-v-Esc DE analysis, with genes enriched in “amoeboid” cells shown as positive on the i x /i -axis. b D, /b Volcano plot for Amo-v-Epi DE analysis, with genes enriched in “amoeboid” cells shown as positive on the i x /i -axis. For B–D, significantly DE genes with an adjusted i P /i value 0.05 are shown in light blue, with no minimum value for log sub /sub FC. b E, /b Expression of EMT markers for epithelial, leaving, and amoeboid cells shown via dot plot. b F, /b Expression of cancer stem cell markers for epithelial, leaving, and amoeboid cells shown via dot plot.
Publisher: American Association for Cancer Research (AACR)
Date: 17-07-2023
DOI: 10.1158/2767-9764.23696152
Abstract: Supplementary Figure 8 shows a summary of GSEA pathway analyses using Hallmarks gene set showing top 25 enriched pathways for each comparison
Publisher: American Association for Cancer Research (AACR)
Date: 17-07-2023
DOI: 10.1158/2767-9764.23696155
Abstract: Supplementary Figure 7 shows metadata for isolated cells
Publisher: American Association for Cancer Research (AACR)
Date: 11-08-2023
DOI: 10.1158/2767-9764.23932481.V1
Abstract: Single-cell tagging and isolation based on morphologic and positional data. b A, /b Sphericity and distance to spheroid values are generated for each invading cell surrounding the spheroid. Left panel depicts maximum intensity projections for three invading cells (Fluo) of different sphericities, following cell surface segmentation (Fluo + Surf), and following surface pseudocoloring according to sphericity value (Surf + Pseudo). Right panels illustrate the smoothed spheroid surface (red) computed to calculate the nearest distance (dashed arrow) to the spheroid edge from in idual cells. b B, /b Left: The three photoconverted cell types “Epithelial,” “Escaping,” and “Amoeboid.” Middle: Representative images of the photoconversion of the three cell types and corresponding flow cytometry scatter plots of Dendra2 red versus green fluorescence obtained during single-cell sorting. Right: The number of cells photoconverted and recovered via single-cell sorting for escaping and amoeboid cells. b C, /b Sorting yields expressed as a percentage of photoconverted cells for escaping and amoeboid cells.
Publisher: American Association for Cancer Research (AACR)
Date: 11-08-2023
DOI: 10.1158/2767-9764.23932463
Abstract: Supplementary Figure 2 shows clustering of 44 single cells and their gene expression heat map based on 230 differentially expressed genes
Publisher: American Association for Cancer Research (AACR)
Date: 11-08-2023
DOI: 10.1158/2767-9764.23932433.V1
Abstract: Supplementary Table 3 shows Differentially expressed genes for escaping vs. epithelial analysis
Publisher: American Association for Cancer Research (AACR)
Date: 11-08-2023
DOI: 10.1158/2767-9764.23932421
Abstract: Supplementary Video 1A shows representative daily images of a vehicle-treated spheroid. Scale bar: 200 µm, number at top left: days post-implantation.
Publisher: American Association for Cancer Research (AACR)
Date: 11-08-2023
DOI: 10.1158/2767-9764.23932466
Abstract: Supplementary Figure 1 shows data generation for the photoconvertible spheroid assay
Publisher: American Association for Cancer Research (AACR)
Date: 11-08-2023
DOI: 10.1158/2767-9764.23932460.V1
Abstract: Supplementary Figure 3 shows validation of OPRD1 and SLC19A1 protein expression
Publisher: American Association for Cancer Research (AACR)
Date: 11-08-2023
DOI: 10.1158/2767-9764.23932454.V1
Abstract: Supplementary Figure 5 shows an Escaping cell cohort gallery
Publisher: American Association for Cancer Research (AACR)
Date: 11-08-2023
DOI: 10.1158/2767-9764.23932424
Abstract: Supplementary Table 6 shows Sorting events for each cell included in the final data set
Publisher: American Association for Cancer Research (AACR)
Date: 17-07-2023
DOI: 10.1158/2767-9764.23696167.V1
Abstract: Supplementary Figure 3 shows validation of OPRD1 and SLC19A1 protein expression
Publisher: American Association for Cancer Research (AACR)
Date: 11-08-2023
DOI: 10.1158/2767-9764.23932469
Abstract: SSZ treatment slows spheroid growth but induces tumor escape by elongated cells. b A, /b Representative maximum intensity projections of imaged hemispheroids for vehicle- and SSZ-treated spheroids. Scale bar, 200 μm. b B, /b Relative spheroid volume for vehicle- and SSZ-treated spheroids. b C, /b Cumulative escaped cells per surface area of imaged hemispheroid. b D, /b Mean escaped cell sphericity per day. B–D, Shaded areas represent SEM.
Publisher: American Association for Cancer Research (AACR)
Date: 11-08-2023
DOI: 10.1158/2767-9764.23932427
Abstract: Supplementary Table 5 shows Differentially expressed genes for epithelial vs. amoeboid analysis
Publisher: American Association for Cancer Research (AACR)
Date: 11-08-2023
DOI: 10.1158/2767-9764.23932412.V1
Abstract: Supplementary Video 2B shows representative images of a Sulfasalazine-treated spheroid. Scale bar: 200 µm, number at top left: days post-implantation.
Publisher: American Association for Cancer Research (AACR)
Date: 17-07-2023
DOI: 10.1158/2767-9764.23696146.V1
Abstract: Supplementary Table 1 shows Cell type isolated, age (days) and number of cells sorted for each spheroid included in the study
Publisher: American Association for Cancer Research (AACR)
Date: 17-07-2023
DOI: 10.1158/2767-9764.23696167
Abstract: Supplementary Figure 3 shows validation of OPRD1 and SLC19A1 protein expression
Publisher: American Association for Cancer Research (AACR)
Date: 17-07-2023
DOI: 10.1158/2767-9764.23696125.V1
Abstract: Supplementary Video 1B shows representative images of a naltrindole-treated spheroid. Scale bar: 200 µm, number at top left: days post-implantation.
Publisher: American Association for Cancer Research (AACR)
Date: 17-07-2023
DOI: 10.1158/2767-9764.23696125
Abstract: Supplementary Video 1B shows representative images of a naltrindole-treated spheroid. Scale bar: 200 µm, number at top left: days post-implantation.
Publisher: American Association for Cancer Research (AACR)
Date: 17-07-2023
DOI: 10.1158/2767-9764.23696173.V1
Abstract: Supplementary Figure 1 shows data generation for the photoconvertible spheroid assay
Publisher: American Association for Cancer Research (AACR)
Date: 17-07-2023
DOI: 10.1158/2767-9764.23696128
Abstract: Supplementary Video 1A shows representative daily images of a vehicle-treated spheroid. Scale bar: 200 µm, number at top left: days post-implantation.
Publisher: American Association for Cancer Research (AACR)
Date: 11-08-2023
DOI: 10.1158/2767-9764.23932445.V1
Abstract: Supplementary Figure 8 shows a summary of GSEA pathway analyses using Hallmarks gene set showing top 25 enriched pathways for each comparison
Publisher: American Association for Cancer Research (AACR)
Date: 11-08-2023
DOI: 10.1158/2767-9764.23932439.V1
Abstract: Supplementary Table 1 shows Cell type isolated, age (days) and number of cells sorted for each spheroid included in the study
Publisher: Wiley
Date: 22-11-2019
Publisher: American Association for Cancer Research (AACR)
Date: 17-07-2023
DOI: 10.1158/2767-9764.23696161
Abstract: Supplementary Figure 5 shows an Escaping cell cohort gallery
Publisher: American Association for Cancer Research (AACR)
Date: 17-07-2023
DOI: 10.1158/2767-9764.23696119.V1
Abstract: Supplementary Video 2B shows representative images of a Sulfasalazine-treated spheroid. Scale bar: 200 µm, number at top left: days post-implantation.
Publisher: American Association for Cancer Research (AACR)
Date: 17-07-2023
DOI: 10.1158/2767-9764.23696131.V1
Abstract: Supplementary Table 6 shows Sorting events for each cell included in the final data set
Publisher: American Association for Cancer Research (AACR)
Date: 17-07-2023
DOI: 10.1158/2767-9764.23696164
Abstract: Supplementary Figure 4 shows an Epithelial cohort gallery
Publisher: American Association for Cancer Research (AACR)
Date: 11-08-2023
DOI: 10.1158/2767-9764.23932481
Abstract: Single-cell tagging and isolation based on morphologic and positional data. b A, /b Sphericity and distance to spheroid values are generated for each invading cell surrounding the spheroid. Left panel depicts maximum intensity projections for three invading cells (Fluo) of different sphericities, following cell surface segmentation (Fluo + Surf), and following surface pseudocoloring according to sphericity value (Surf + Pseudo). Right panels illustrate the smoothed spheroid surface (red) computed to calculate the nearest distance (dashed arrow) to the spheroid edge from in idual cells. b B, /b Left: The three photoconverted cell types “Epithelial,” “Escaping,” and “Amoeboid.” Middle: Representative images of the photoconversion of the three cell types and corresponding flow cytometry scatter plots of Dendra2 red versus green fluorescence obtained during single-cell sorting. Right: The number of cells photoconverted and recovered via single-cell sorting for escaping and amoeboid cells. b C, /b Sorting yields expressed as a percentage of photoconverted cells for escaping and amoeboid cells.
Publisher: American Association for Cancer Research (AACR)
Date: 17-07-2023
DOI: 10.1158/2767-9764.23696122
Abstract: Supplementary Video 2A shows representative images of a vehicle-treated spheroid. Scale bar: 200 µm, number at top left: days post-implantation.
Publisher: American Association for Cancer Research (AACR)
Date: 11-08-2023
DOI: 10.1158/2767-9764.23932472
Abstract: Inhibition of OPRD1 reduces melanoma spheroid growth and invasion. b A, /b Representative maximum intensity projections of imaged hemispheroids for vehicle- and naltrindole-treated spheroids, scale bar 200 μm. b B, /b Relative spheroid volume over time for vehicle- and naltrindole-treated cells. Shaded areas represent SEM. b C, /b Cumulative number of escaped cells over time normalized to spheroid surface area. B and C, Datapoints represent the mean of eight replicates.
Publisher: American Association for Cancer Research (AACR)
Date: 17-07-2023
DOI: 10.1158/2767-9764.23696164.V1
Abstract: Supplementary Figure 4 shows an Epithelial cohort gallery
Publisher: American Association for Cancer Research (AACR)
Date: 11-08-2023
DOI: 10.1158/2767-9764.23932430
Abstract: Supplementary Table 4 shows Differentially expressed genes for amoeboid vs. escaping analysis
Publisher: American Association for Cancer Research (AACR)
Date: 11-08-2023
DOI: 10.1158/2767-9764.23932478.V1
Abstract: Clustering of single cells and their gene expression heat map based on 462 DE genes. b A, /b UMAP clustering of 35 cells based on expression of 462 DE genes. b B, /b Unsupervised hierarchical clustering of 35 cells based on the expression of 462 DE genes. Cells initially erge into two groups, epithelial (L1) and invasive (R1), before further erging into escaping (R2) and amoeboid (R3). b C, /b Heat map of 462 DE genes for single cells arranged according to the unsupervised clustering in B.
Publisher: American Association for Cancer Research (AACR)
Date: 11-08-2023
DOI: 10.1158/2767-9764.23932475
Abstract: Gene expression analysis for epithelial, escaping, and amoeboid cells. b A, /b Top: 462 genes were DE, 144 for escaping versus epithelial (Esc-v-Epi), 22 for amoeboid versus escaping (Amo-v-Esc), and 422 for amoeboid versus epithelial (Amo-v-Epi). Bottom: Venn diagram of the number of DE genes for the three pairwise comparisons. b B, /b Volcano plot for Esc-v-Epi DE analysis, with genes enriched in “escaping” cells shown as positive on the i x /i -axis. b C, /b Volcano plot for Amo-v-Esc DE analysis, with genes enriched in “amoeboid” cells shown as positive on the i x /i -axis. b D, /b Volcano plot for Amo-v-Epi DE analysis, with genes enriched in “amoeboid” cells shown as positive on the i x /i -axis. For B–D, significantly DE genes with an adjusted i P /i value 0.05 are shown in light blue, with no minimum value for log sub /sub FC. b E, /b Expression of EMT markers for epithelial, leaving, and amoeboid cells shown via dot plot. b F, /b Expression of cancer stem cell markers for epithelial, leaving, and amoeboid cells shown via dot plot.
Publisher: American Association for Cancer Research (AACR)
Date: 11-08-2023
DOI: 10.1158/2767-9764.23932433
Abstract: Supplementary Table 3 shows Differentially expressed genes for escaping vs. epithelial analysis
Publisher: American Association for Cancer Research (AACR)
Date: 11-08-2023
DOI: 10.1158/2767-9764.23932484.V1
Abstract: Methodologic pipeline for single-cell isolation of live escaping tumor cells. b , /b A clonal population of tumor cells are expanded from a single cell. b , /b Photoconvertible cells are seeded in ULA plates to generate spheroids. b , /b Spheroids are embedded within a 3D collagen matrix and imaged over time by light microscopy. b , /b Image analysis and extraction of quantitative metrics can be used to select cells of interest. b , /b Single cells of interest are photoconverted using a 405 nm laser. b , /b Collagen matrix is enzymatically digested to provide a solution of suspended single cells. b , /b Cells are sorted via FACS. b , /b Cells can be further cultured or immediately analyzed.
Publisher: American Association for Cancer Research (AACR)
Date: 11-08-2023
DOI: 10.1158/2767-9764.23932436.V1
Abstract: Supplementary Table 2 shows Transcript counts for common housekeeping genes, with cells excluded from the study due to low housekeeping expression highlighted in yellow
Publisher: American Association for Cancer Research (AACR)
Date: 11-08-2023
DOI: 10.1158/2767-9764.23932478
Abstract: Clustering of single cells and their gene expression heat map based on 462 DE genes. b A, /b UMAP clustering of 35 cells based on expression of 462 DE genes. b B, /b Unsupervised hierarchical clustering of 35 cells based on the expression of 462 DE genes. Cells initially erge into two groups, epithelial (L1) and invasive (R1), before further erging into escaping (R2) and amoeboid (R3). b C, /b Heat map of 462 DE genes for single cells arranged according to the unsupervised clustering in B.
Publisher: American Association for Cancer Research (AACR)
Date: 11-08-2023
DOI: 10.1158/2767-9764.23932436
Abstract: Supplementary Table 2 shows Transcript counts for common housekeeping genes, with cells excluded from the study due to low housekeeping expression highlighted in yellow
Publisher: American Association for Cancer Research (AACR)
Date: 11-08-2023
DOI: 10.1158/2767-9764.23932451.V1
Abstract: Supplementary Figure 6 shows an Amoeboid cell cohort gallery
Publisher: American Association for Cancer Research (AACR)
Date: 11-08-2023
DOI: 10.1158/2767-9764.23932439
Abstract: Supplementary Table 1 shows Cell type isolated, age (days) and number of cells sorted for each spheroid included in the study
Publisher: American Association for Cancer Research (AACR)
Date: 17-07-2023
DOI: 10.1158/2767-9764.23696122.V1
Abstract: Supplementary Video 2A shows representative images of a vehicle-treated spheroid. Scale bar: 200 µm, number at top left: days post-implantation.
Publisher: American Association for Cancer Research (AACR)
Date: 17-07-2023
DOI: 10.1158/2767-9764.23696134
Abstract: Supplementary Table 5 shows Differentially expressed genes for epithelial vs. amoeboid analysis
Publisher: American Association for Cancer Research (AACR)
Date: 11-08-2023
DOI: 10.1158/2767-9764.23932442.V1
Abstract: Supplementary Figure 9 shows Escaped cell analyses for naltrindole and sulfasalazine spheroid assays
Publisher: American Association for Cancer Research (AACR)
Date: 11-08-2023
DOI: 10.1158/2767-9764.23932421.V1
Abstract: Supplementary Video 1A shows representative daily images of a vehicle-treated spheroid. Scale bar: 200 µm, number at top left: days post-implantation.
Publisher: American Association for Cancer Research (AACR)
Date: 17-07-2023
DOI: 10.1158/2767-9764.23696137
Abstract: Supplementary Table 4 shows Differentially expressed genes for amoeboid vs. escaping analysis
Publisher: American Association for Cancer Research (AACR)
Date: 17-07-2023
DOI: 10.1158/2767-9764.23696155.V1
Abstract: Supplementary Figure 7 shows metadata for isolated cells
Publisher: American Association for Cancer Research (AACR)
Date: 11-08-2023
DOI: 10.1158/2767-9764.23932463.V1
Abstract: Supplementary Figure 2 shows clustering of 44 single cells and their gene expression heat map based on 230 differentially expressed genes
Publisher: American Association for Cancer Research (AACR)
Date: 17-07-2023
DOI: 10.1158/2767-9764.23696170
Abstract: Supplementary Figure 2 shows clustering of 44 single cells and their gene expression heat map based on 230 differentially expressed genes
Publisher: American Association for Cancer Research (AACR)
Date: 17-07-2023
DOI: 10.1158/2767-9764.23696134.V1
Abstract: Supplementary Table 5 shows Differentially expressed genes for epithelial vs. amoeboid analysis
Publisher: American Association for Cancer Research (AACR)
Date: 17-07-2023
DOI: 10.1158/2767-9764.23696173
Abstract: Supplementary Figure 1 shows data generation for the photoconvertible spheroid assay
Publisher: American Association for Cancer Research (AACR)
Date: 17-07-2023
DOI: 10.1158/2767-9764.23696131
Abstract: Supplementary Table 6 shows Sorting events for each cell included in the final data set
Publisher: eLife Sciences Publications, Ltd
Date: 04-2020
DOI: 10.7554/ELIFE.53308
Abstract: Mechanoelectrical transduction is a cellular signalling pathway where physical stimuli are converted into electro-chemical signals by mechanically activated ion channels. We describe here the presence of mechanically activated currents in melanoma cells that are dependent on TMEM87a, which we have renamed Elkin1. Heterologous expression of this protein in PIEZO1-deficient cells, that exhibit no baseline mechanosensitivity, is sufficient to reconstitute mechanically activated currents. Melanoma cells lacking functional Elkin1 exhibit defective mechanoelectrical transduction, decreased motility and increased dissociation from organotypic spheroids. By analysing cell adhesion properties, we demonstrate that Elkin1 deletion is associated with increased cell-substrate adhesion and decreased homotypic cell-cell adhesion strength. We therefore conclude that Elkin1 supports a PIEZO1-independent mechanoelectrical transduction pathway and modulates cellular adhesions and regulates melanoma cell migration and cell-cell interactions.
Publisher: American Association for Cancer Research (AACR)
Date: 11-08-2023
DOI: 10.1158/2767-9764.23932484
Abstract: Methodologic pipeline for single-cell isolation of live escaping tumor cells. b , /b A clonal population of tumor cells are expanded from a single cell. b , /b Photoconvertible cells are seeded in ULA plates to generate spheroids. b , /b Spheroids are embedded within a 3D collagen matrix and imaged over time by light microscopy. b , /b Image analysis and extraction of quantitative metrics can be used to select cells of interest. b , /b Single cells of interest are photoconverted using a 405 nm laser. b , /b Collagen matrix is enzymatically digested to provide a solution of suspended single cells. b , /b Cells are sorted via FACS. b , /b Cells can be further cultured or immediately analyzed.
Publisher: American Association for Cancer Research (AACR)
Date: 17-07-2023
DOI: 10.1158/2767-9764.23696140.V1
Abstract: Supplementary Table 3 shows Differentially expressed genes for escaping vs. epithelial analysis
Publisher: American Association for Cancer Research (AACR)
Date: 17-07-2023
DOI: 10.1158/2767-9764.23696137.V1
Abstract: Supplementary Table 4 shows Differentially expressed genes for amoeboid vs. escaping analysis
Publisher: American Association for Cancer Research (AACR)
Date: 11-08-2023
DOI: 10.1158/2767-9764.23932442
Abstract: Supplementary Figure 9 shows Escaped cell analyses for naltrindole and sulfasalazine spheroid assays
Publisher: American Association for Cancer Research (AACR)
Date: 17-07-2023
DOI: 10.1158/2767-9764.23696161.V1
Abstract: Supplementary Figure 5 shows an Escaping cell cohort gallery
Publisher: American Association for Cancer Research (AACR)
Date: 11-08-2023
DOI: 10.1158/2767-9764.C.6745837.V2
Abstract: Solid cancer cells escape the primary tumor mass by transitioning from an epithelial-like state to an invasive migratory state. As they escape, metastatic cancer cells employ interchangeable modes of invasion, transitioning between fibroblast-like mesenchymal movement to amoeboid migration, where cells display a rounded morphology and navigate the extracellular matrix in a protease-independent manner. However, the gene transcripts that orchestrate the switch between epithelial, mesenchymal, and amoeboid states remain incompletely mapped, mainly due to a lack of methodologies that allow the direct comparison of the transcriptomes of spontaneously invasive cancer cells in distinct migratory states. Here, we report a novel single-cell isolation technique that provides detailed three-dimensional data on melanoma growth and invasion, and enables the isolation of live, spontaneously invasive cancer cells with distinct morphologies and invasion parameters. Via the expression of a photoconvertible fluorescent protein, compact epithelial-like cells at the periphery of a melanoma mass, elongated cells in the process of leaving the mass, and rounded amoeboid cells invading away from the mass were tagged, isolated, and subjected to single-cell RNA sequencing. A total of 462 differentially expressed genes were identified, from which two candidate proteins were selected for further pharmacologic perturbation, yielding striking effects on tumor escape and invasion, in line with the predictions from the transcriptomics data. This work describes a novel, adaptable, and readily implementable method for the analysis of the earliest phases of tumor escape and metastasis, and its application to the identification of genes underpinning the invasiveness of malignant melanoma. Significance: This work describes a readily implementable method that allows for the isolation of in idual live tumor cells of interest for downstream analyses, and provides the single-cell transcriptomes of melanoma cells at distinct invasive states, both of which open avenues for in-depth investigations into the transcriptional regulation of the earliest phases of metastasis. /
Publisher: American Association for Cancer Research (AACR)
Date: 11-08-2023
DOI: 10.1158/2767-9764.23932445
Abstract: Supplementary Figure 8 shows a summary of GSEA pathway analyses using Hallmarks gene set showing top 25 enriched pathways for each comparison
Publisher: American Association for Cancer Research (AACR)
Date: 17-07-2023
DOI: 10.1158/2767-9764.23696158.V1
Abstract: Supplementary Figure 6 shows an Amoeboid cell cohort gallery
Publisher: American Association for Cancer Research (AACR)
Date: 17-07-2023
DOI: 10.1158/2767-9764.C.6745837.V1
Abstract: Abstract Solid cancer cells escape the primary tumour mass by transitioning from an epithelial-like state to an invasive migratory state. As they escape, metastatic cancer cells employ interchangeable modes of invasion, transitioning between fibroblast-like mesenchymal movement to amoeboid migration, where cells display a rounded morphology and navigate the extracellular matrix in a protease-independent manner. However, the gene transcripts that orchestrate the switch between epithelial, mesenchymal and amoeboid states remain incompletely mapped, mainly due to a lack of methodologies that allow the direct comparison of the transcriptomes of spontaneously invasive cancer cells in distinct migratory states. Here, we report a novel single cell isolation technique that provides detailed 3D data on melanoma growth and invasion, and enables the isolation of live, spontaneously invasive cancer cells with distinct morphologies and invasion parameters. Via the expression of a photoconvertible fluorescent protein, compact epithelial-like cells at the periphery of a melanoma mass, elongated cells in the process of leaving the mass, and rounded amoeboid cells invading away from the mass were tagged, isolated and subjected to single cell RNA-sequencing. 462 differentially expressed genes were identified, from which two candidate proteins were selected for further pharmacological perturbation, yielding striking effects on tumour escape and invasion, in line with the predictions from the transcriptomics data. This work describes a novel, adaptable and readily implementable method for the analysis of the earliest phases of tumour escape and metastasis, and its application to the identification of genes underpinning the invasiveness of malignant melanoma. /
Publisher: American Association for Cancer Research (AACR)
Date: 11-08-2023
DOI: 10.1158/2767-9764.23932448
Abstract: Supplementary Figure 7 shows metadata for isolated cells
Publisher: American Association for Cancer Research (AACR)
Date: 11-08-2023
DOI: 10.1158/2767-9764.C.6745837
Abstract: Solid cancer cells escape the primary tumor mass by transitioning from an epithelial-like state to an invasive migratory state. As they escape, metastatic cancer cells employ interchangeable modes of invasion, transitioning between fibroblast-like mesenchymal movement to amoeboid migration, where cells display a rounded morphology and navigate the extracellular matrix in a protease-independent manner. However, the gene transcripts that orchestrate the switch between epithelial, mesenchymal, and amoeboid states remain incompletely mapped, mainly due to a lack of methodologies that allow the direct comparison of the transcriptomes of spontaneously invasive cancer cells in distinct migratory states. Here, we report a novel single-cell isolation technique that provides detailed three-dimensional data on melanoma growth and invasion, and enables the isolation of live, spontaneously invasive cancer cells with distinct morphologies and invasion parameters. Via the expression of a photoconvertible fluorescent protein, compact epithelial-like cells at the periphery of a melanoma mass, elongated cells in the process of leaving the mass, and rounded amoeboid cells invading away from the mass were tagged, isolated, and subjected to single-cell RNA sequencing. A total of 462 differentially expressed genes were identified, from which two candidate proteins were selected for further pharmacologic perturbation, yielding striking effects on tumor escape and invasion, in line with the predictions from the transcriptomics data. This work describes a novel, adaptable, and readily implementable method for the analysis of the earliest phases of tumor escape and metastasis, and its application to the identification of genes underpinning the invasiveness of malignant melanoma. Significance: This work describes a readily implementable method that allows for the isolation of in idual live tumor cells of interest for downstream analyses, and provides the single-cell transcriptomes of melanoma cells at distinct invasive states, both of which open avenues for in-depth investigations into the transcriptional regulation of the earliest phases of metastasis. /
Location: United Kingdom of Great Britain and Northern Ireland
Location: Indonesia
No related grants have been discovered for Jacqueline Tearle.