BAG6 restricts pancreatic cancer progression by suppressing the release of IL33-presenting extracellular vesicles and the activation of mast cells

Bilal Alashkar Alhamwe; Viviane Ponath; Fahd Alhamdan; Bastian Dörsam; Clara Landwehr; Manuel Linder; Kim Pauck; Sarah Miethe; Holger Garn; Florian Finkernagel; Anna Brichkina; Matthias Lauth; Dinesh Kumar Tiwari; Malte Buchholz; Daniel Bachurski; Sabrina Elmshäuser; Andrea Nist; Thorsten Stiewe; Lisa Pogge von Strandmann; Witold Szymański; Vanessa Beutgen; Johannes Graumann; Julia Teply-Szymanski; Corinna Keber; Carsten Denkert; Ralf Jacob; Christian Preußer; Elke Pogge von Strandmann

doi:10.1038/s41423-024-01195-1

BAG6 restricts pancreatic cancer progression by suppressing the release of IL33-presenting extracellular vesicles and the activation of mast cells

Cell Mol Immunol. 2024 Aug;21(8):918-931. doi: 10.1038/s41423-024-01195-1. Epub 2024 Jun 28.

Authors

Bilal Alashkar Alhamwe^{1

2}, Viviane Ponath^{1

2}, Fahd Alhamdan^{3

4}, Bastian Dörsam^{1

2}, Clara Landwehr^{1

2}, Manuel Linder^{1

2}, Kim Pauck⁵, Sarah Miethe⁵, Holger Garn⁵, Florian Finkernagel^{1

6}, Anna Brichkina^{7

8}, Matthias Lauth⁷, Dinesh Kumar Tiwari⁷, Malte Buchholz⁷, Daniel Bachurski⁹, Sabrina Elmshäuser¹⁰, Andrea Nist¹⁰, Thorsten Stiewe^{10

11}, Lisa Pogge von Strandmann^{1

2}, Witold Szymański¹², Vanessa Beutgen¹², Johannes Graumann¹², Julia Teply-Szymanski¹³, Corinna Keber¹³, Carsten Denkert¹³, Ralf Jacob¹⁴, Christian Preußer^{1

2}, Elke Pogge von Strandmann^{15

16}

Affiliations

¹ Institute for Tumor Immunology, Philipps-University, 35043, Marburg, Germany.
² Core Facility Extracellular Vesicles, Philipps-University, 35043, Marburg, Germany.
³ Department of Anesthesiology, Critical Care, and Pain Medicine, Cardiac Anesthesia Division, Boston Children's Hospital, Boston, USA.
⁴ Department of Immunology and Anaesthesia, Harvard Medical School, Boston, MA, USA.
⁵ Translational Inflammation Research Division & Core Facility for Single Cell Multiomics, Philipps-University, 35043, Marburg, Germany.
⁶ Core Facility Bioinformatics, Philipps-University, 35043, Marburg, Germany.
⁷ Clinic for Gastroenterology, Endocrinology and Metabolism; Center for Tumor and Immune Biology, Philipps-University, 35043, Marburg, Germany.
⁸ Institute of Systems Immunology, Philipps-University, 35043, Marburg, Germany.
⁹ Cluster of Excellence on Cellular Stress Responses in Aging-Associated Diseases (CECAD), University of Cologne, Cologne, Germany.
¹⁰ Institute of Molecular Oncology and Genomics Core Facility, Member of the German Center for Lung Research (DZL), Philipps-University, 35043, Marburg, Germany.
¹¹ Institute of Lung Health, Justus Liebig University, 35392, Giessen, Germany.
¹² Institute of Translational Proteomics & Core Facility Translational Proteomics, Biochemical/Pharmacological Centre, Philipps-University, 35043, Marburg, Germany.
¹³ Institute of Pathology, Philipps-University Marburg and University Hospital Marburg (UKGM), Marburg, Germany.
¹⁴ Department of Cell Biology and Cell Pathology, Philipps-University, 35043, Marburg, Germany.
¹⁵ Institute for Tumor Immunology, Philipps-University, 35043, Marburg, Germany. poggevon@staff.uni-marburg.de.
¹⁶ Core Facility Extracellular Vesicles, Philipps-University, 35043, Marburg, Germany. poggevon@staff.uni-marburg.de.

Abstract

Recent studies reveal a critical role of tumor cell-released extracellular vesicles (EVs) in pancreatic cancer (PC) progression. However, driver genes that direct EV function, the EV-recipient cells, and their cellular response to EV uptake remain to be identified. Therefore, we studied the role of Bcl-2-associated-anthanogene 6 (BAG6), a regulator of EV biogenesis for cancer progression. We used a Cre recombinase/LoxP-based reporter system in combination with single-cell RNA sequencing to monitor in vivo EV uptake and tumor microenvironment (TME) changes in mouse models for pancreatic ductal adenocarcinoma (PDAC) in a Bag6 pro- or deficient background. In vivo data were validated using mouse and human organoids and patient samples. Our data demonstrated that Bag6-deficient subcutaneous and orthotopic PDAC tumors accelerated tumor growth dependent on EV release. Mechanistically, this was attributed to mast cell (MC) activation via EV-associated IL33. Activated MCs promoted tumor cell proliferation and altered the composition of the TME affecting fibroblast polarization and immune cell infiltration. Tumor cell proliferation and fibroblast polarization were mediated via the MC secretome containing high levels of PDGF and CD73. Patients with high BAG6 gene expression and high protein plasma level have a longer overall survival indicating clinical relevance. The current study revealed a so far unknown tumor-suppressing activity of BAG6 in PDAC. Bag6-deficiency allowed the release of EV-associated IL33 which modulate the TME via MC activation promoting aggressive tumor growth. MC depletion using imatinib diminished tumor growth providing a scientific rationale to consider imatinib for patients stratified with low BAG6 expression and high MC infiltration. EVs derived from BAG6-deficient pancreatic cancer cells induce MC activation via IL33/Il1rl1. The secretome of activated MCs induces tumor proliferation and changes in the TME, particularly shifting fibroblasts into an inflammatory cancer-associated fibroblast (iCAF) phenotype. Blocking EVs or depleting MCs restricts tumor growth.

Keywords: BAG6; EVs; Mast cells; Pancreatic cancer.

MeSH terms

Animals
Carcinoma, Pancreatic Ductal* / genetics
Carcinoma, Pancreatic Ductal* / immunology
Carcinoma, Pancreatic Ductal* / pathology
Cell Line, Tumor
Cell Proliferation
Disease Progression*
Extracellular Vesicles* / metabolism
Humans
Interleukin-33* / genetics
Interleukin-33* / metabolism
Mast Cells* / immunology
Mast Cells* / metabolism
Mice
Mice, Inbred C57BL
Pancreatic Neoplasms* / immunology
Pancreatic Neoplasms* / pathology
Tumor Microenvironment*

Substances

Interleukin-33
BAG6 protein, human
Bag6 protein, mouse
IL33 protein, human
Il33 protein, mouse

Abstract

MeSH terms

Substances

Grants and funding