The Gustatory Sensory G-Protein GNAT3 Suppresses Pancreatic Cancer Progression in Mice
| Autor: | Megan T. Hoffman, Samantha B. Kemp, Daniel J. Salas-Escabillas, Yaqing Zhang, Nina G. Steele, Stephanie The, Daniel Long, Simone Benitz, Wei Yan, Robert F. Margolskee, Filip Bednar, Marina Pasca di Magliano, Hui-Ju Wen, Howard C. Crawford |
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| Rok vydání: | 2021 |
| Předmět: |
0301 basic medicine
KrasG12D/+ Ptf1aCreERT/+ MDSC CK19 cytokeratin 19 medicine.disease_cause Ptf1aCre/+ KCERT 0302 clinical medicine MTC metaplastic tuft cell Original Research gMDSC granulocytic myeloid-derived suppressor cell education.field_of_study KC KrasG12D/+ Gastroenterology ELISA enzyme-linked immunosorbent assay PDA pancreatic ductal adenocarcinoma CXCL2 CXCL1 DCLK1 doublecortin-like kinase 1 medicine.anatomical_structure PTOM pancreatic progenitor and tumor organoid media TAM tumor-associated macrophage 030211 gastroenterology & hepatology Tuft cell Pancreas NK natural killer IHC immunohistochemistry Population PBS phosphate-buffered saline Tuft Cell CC3 cleaved caspase 3 Biology TRPM5 transient receptor potential cation channel subfamily M member 5 ADM acinar-to-ductal metaplasia 03 medical and health sciences FBS fetal bovine serum MDSC myeloid-derived suppressor cell Pancreatic cancer UMAP Uniform Manifold Approximation and Projection medicine Humans mMDSC monocytic myeloid-derived suppressor cell education Hepatology GNAT3 α-gustducin HBSS Hank’s balanced salt solution PanIN pancreatic intraepithelial neoplasia medicine.disease WT wild-type Pancreatic Neoplasms 030104 developmental biology Cancer research BSA bovine serum albumin Cytokine secretion Carcinogenesis |
| Zdroj: | Cellular and Molecular Gastroenterology and Hepatology |
| ISSN: | 2352-345X |
| DOI: | 10.1016/j.jcmgh.2020.08.011 |
| Popis: | Background & Aims Pancreatic ductal adenocarcinoma (PDA) initiation and progression are accompanied by an immunosuppressive inflammatory response. Here, we evaluated the immunomodulatory role of chemosensory signaling in metaplastic tuft cells (MTCs) by analyzing the role of GNAT3, a gustatory pathway G-protein expressed by MTCs, during PDA progression. Methods Gnat3-null (Gnat3-/-) mice were crossbred with animals harboring a Cre-inducible KrasLSL-G12D/+ allele with either Ptf1aCre/+ (KC) or tamoxifen-inducible Ptf1aCreERT/+ (KCERT) mice to drive oncogenic KRAS expression in the pancreas. Ex vivo organoid conditioned medium generated from KC and Gnat3-/-;KC acinar cells was analyzed for cytokine secretion. Experimental pancreatitis was induced in KCERT and Gnat3-/-;KCERT mice to accelerate tumorigenesis, followed by analysis using mass cytometry and single-cell RNA sequencing. To study PDA progression, KC and Gnat3-/-;KC mice were aged to morbidity or 52 weeks. Results Ablation of Gnat3 in KC organoids increased release of tumor-promoting cytokines in conditioned media, including CXCL1 and CXCL2. Analysis of Gnat3-/-;KCERT pancreata found altered expression of immunomodulatory genes in Cxcr2 expressing myeloid-derived suppressor cells (MDSCs) and an increased number of granulocytic MDSCs, a subset of tumor promoting MDSCs. Importantly, expression levels of CXCL1 and CXCL2, known ligands for CXCR2, were also elevated in Gnat3-/-;KCERT pancreata. Consistent with the tumor-promoting role of MDSCs, aged Gnat3-/-;KC mice progressed more rapidly to metastatic carcinoma compared with KC controls. Conclusions Compromised gustatory sensing, achieved by Gnat3 ablation, enhanced the CXCL1/2–CXCR2 axis to alter the MDSC population and promoted the progression of metastatic PDA. Graphical abstract |
| Databáze: | OpenAIRE |
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