27787a361288f268ad20e9aecda8ae16 pmc_9139899.pdf 8c1bbbcc9cefeb9e8e11be95c1655d2551eea0c8 pmc_9139899.pdf a7a038dcd4ab4ab0ba9df88a833a08e17dde3f82bc43a767fe9dfcf243d48c6e pmc_9139899.pdf Title: GDF15 Is an Eribulin Response Biomarker also Required for Survival of DTP Breast Cancer Cells Subject: Drug tolerant persister (DTP) cells enter into a reversible slow-cycling state after drug treatment. We performed proteomic characterization of the breast cancer (BC) DTP cell secretome after eribulin treatment. We showed that the growth differentiation factor 15 (GDF15) is a protein significantly over-secreted upon eribulin treatment. The biomarker potential of GDF15 was confirmed in 3D-cell culture models using BC cells lines and PDXs, as well as in a TNBC in vivo model. We also found that GDF15 is required for survival of DTP cells. Direct participation of GDF15 and its receptor GFRAL in eribulin-induction of DTPs was established by the enhanced cell killing of DTPs by eribulin seen under GDF15 and GFRAL loss of function assays. Finally, we showed that combination therapy of eribulin plus an anti-GDF15 antibody kills BC-DTP cells. Our results suggest that targeting GDF15 may help eradicate DTP cells and block the onset of acquired resistance. Keywords: drug tolerance; drug tolerant persister (DTP); eribulin; growth differentiation factor 15 (GDF15); breast cancer (BC); secretome Author: Chiara Bellio, Marta Emperador, Pol Castellano, Albert Gris-Oliver, Francesc Canals, Alex Sánchez-Pla, Esther Zamora, Joaquín Arribas, Cristina Saura, Violeta Serra, Josep Tabernero, Bruce A. Littlefield and Josep Villanueva Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Tue May 24 08:38:07 2022 CEST ModDate: Thu May 26 07:53:04 2022 CEST Custom Metadata: no Metadata Stream: no Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 23 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 4459954 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- SSNIUI+VnURWPalladioL-Bold Type 1 Custom yes yes no 10 0 OPZDHV+VnURWPalladioL Type 1 Custom yes yes no 15 0 OQIKBX+URWPalladioL-Bold Type 1 Custom yes yes no 19 0 LDRNOB+URWPalladioL-Roma Type 1 Custom yes yes no 24 0 FERLAG+URWPalladioL-Ital Type 1 Custom yes yes no 28 0 LCJEVJ+TimesNewRomanPS-BoldItalicMT Type 1C WinAnsi yes yes no 47 0 CAMXTK+EURM10 Type 1 Builtin yes yes no 69 0 QMDDXF+CMSY10 Type 1 Builtin yes yes no 76 0 WBVKDJ+URWPalladioL-BoldItal Type 1 Custom yes yes no 116 0 GHCAME+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 151 0 GHCAMG+PalatinoLinotype TrueType WinAnsi yes yes no 154 0 GHCAMI+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 157 0 GHCAME+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 171 0 GHCAMG+PalatinoLinotype TrueType WinAnsi yes yes no 174 0 GHCAMI+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 177 0 Jhove (Rel. 1.28.0, 2023-05-18) Date: 2024-05-18 03:24:58 CEST RepresentationInformation: pmc_9139899.pdf ReportingModule: PDF-hul, Rel. 1.12.4 (2023-03-16) LastModified: 2024-05-18 02:37:09 CEST Size: 4459954 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 400 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: section.1 Item: Title: Material and Methods Destination: section.2 Children: Item: Title: Cell Culture and Breast Cancer PDX Tumor Cells Isolation Destination: subsection.2.1 Item: Title: Three-Dimensional Cell Culture Organoids Destination: subsection.2.2 Item: Title: Drug Treatment (Eribulin, Vinorelbine and Paclitaxel) CellTiter, Cell Counting and Crystal Violet Staining Destination: subsection.2.3 Item: Title: Immunofluorescence and Confocal Microscopy Destination: subsection.2.4 Item: Title: Western Blotting Destination: subsection.2.5 Item: Title: Cell Proliferation Assays: Cell Cycle and PKH26 Staining Destination: subsection.2.6 Item: Title: Secretome Collection Destination: subsection.2.7 Item: Title: Liquid Chromatography Mass Spectrometry Analysis (LC MS) and Protein Identification Destination: subsection.2.8 Item: Title: Protein Identification and Quantitative Differential Analysis Destination: subsection.2.9 Item: Title: siRNA Experiment Destination: subsection.2.10 Item: Title: Anti-GDF15 Experiment Destination: subsection.2.11 Item: Title: In Vivo Experiment Destination: subsection.2.12 Item: Title: Statistical Analysis Destination: subsection.2.13 Item: Title: Results Destination: section.3 Children: Item: Title: Eribulin Treatment Leads to Formation of a Drug-Tolerant Persister (DTP) Subpopulation in Breast Cancer Cell Lines Destination: subsection.3.1 Item: Title: Eribulin-Induced DTP Cells Display a Specific Secretome Destination: subsection.3.2 Item: Title: Secretomes Induced by MTAs in Breast Cancer Cells Are Linked to Their Tubulin-Binding Mechanism Destination: subsection.3.3 Item: Title: GDF15 Secretion Is Strongly Associated with the DTP State Destination: subsection.3.4 Item: Title: GDF15 Is Required for Maintenance of the DTP State in Cells Responding to Eribulin Destination: subsection.3.5 Item: Title: Discussion Destination: section.4 Item: Title: Conclusions Destination: section.5 Item: Title: References Destination: section.5 Info: Title: GDF15 Is an Eribulin Response Biomarker also Required for Survival of DTP Breast Cancer Cells Author: Chiara Bellio, Marta Emperador, Pol Castellano, Albert Gris-Oliver, Francesc Canals, Alex Sánchez-Pla, Esther Zamora, Joaquín Arribas, Cristina Saura, Violeta Serra, Josep Tabernero, Bruce A. Littlefield and Josep Villanueva Subject: Drug tolerant persister (DTP) cells enter into a reversible slow-cycling state after drug treatment. We performed proteomic characterization of the breast cancer (BC) DTP cell secretome after eribulin treatment. We showed that the growth differentiation factor 15 (GDF15) is a protein significantly over-secreted upon eribulin treatment. The biomarker potential of GDF15 was confirmed in 3D-cell culture models using BC cells lines and PDXs, as well as in a TNBC in vivo model. We also found that GDF15 is required for survival of DTP cells. Direct participation of GDF15 and its receptor GFRAL in eribulin-induction of DTPs was established by the enhanced cell killing of DTPs by eribulin seen under GDF15 and GFRAL loss of function assays. Finally, we showed that combination therapy of eribulin plus an anti-GDF15 antibody kills BC-DTP cells. Our results suggest that targeting GDF15 may help eradicate DTP cells and block the onset of acquired resistance. 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