Transformation of primary murine peritoneal mast cells by constitutive KIT activation is accompanied by loss of Cdkn2a/Arf expression

Sandro Capellmann; Roland Sonntag; Herdit Schüler; Steffen K Meurer; Lin Gan; Marlies Kauffmann; Katharina Horn; Hiltrud Königs-Werner; Ralf Weiskirchen; Christian Liedtke; Michael Huber

doi:10.3389/fimmu.2023.1154416

Transformation of primary murine peritoneal mast cells by constitutive KIT activation is accompanied by loss of Cdkn2a/Arf expression

Front Immunol. 2023 Mar 30:14:1154416. doi: 10.3389/fimmu.2023.1154416. eCollection 2023.

Affiliations

¹ Institute of Biochemistry and Molecular Immunology, Medical Faculty, Rheinisch-Westfälische Technische Hochschule (RWTH) Aachen University, Aachen, Germany.
² Department of Internal Medicine III, University Hospital, Rheinisch-Westfälische Technische Hochschule (RWTH) Aachen University, Aachen, Germany.
³ Institute of Human Genetics, Medical Faculty, Rheinisch-Westfälische Technische Hochschule (RWTH) Aachen University, Aachen, Germany.
⁴ Institute of Molecular Pathobiochemistry, Experimental Gene Therapy and Clinical Chemistry (IFMPEGKC), Medical Faculty, Rheinisch-Westfälische Technische Hochschule (RWTH) Aachen University, Aachen, Germany.
⁵ Genomics Facility, Interdisciplinary Center for Clinical Research (IZKF), Medical Faculty, Rheinisch-Westfälische Technische Hochschule (RWTH) Aachen University, Aachen, Germany.
⁶ Electron Microscopy Facility, Institute of Pathology, Medical Faculty, Rheinisch-Westfälische Technische Hochschule (RWTH) Aachen University, Aachen, Germany.

Abstract

Mast cells (MCs) are immune cells of the myeloid lineage distributed in tissues throughout the body. Phenotypically, they are a heterogeneous group characterized by different protease repertoires stored in secretory granules and differential presence of receptors. To adequately address aspects of MC biology either primary MCs isolated from human or mouse tissue or different human MC lines, like HMC-1.1 and -1.2, or rodent MC lines like L138.8A or RBL-2H3 are frequently used. Nevertheless, cellular systems to study MC functions are very limited. We have generated a murine connective tissue-like MC line, termed PMC-306, derived from primary peritoneal MCs (PMCs), which spontaneously transformed. We analyzed PMC-306 cells regarding MC surface receptor expression, effector functions and respective signaling pathways, and found that the cells reacted very similar to primary wildtype (WT) PMCs. In this regard, stimulation with MAS-related G-protein-coupled receptor member B2 (MRGPRB2) ligands induced respective signaling and effector functions. Furthermore, PMC-306 cells revealed significantly accelerated cell cycle progression, which however was still dependent on interleukine 3 (IL-3) and stem cell factor (SCF). Phenotypically, PMC-306 cells adopted an immature connective tissue-like MCs appearance. The observation of cellular transformation was accompanied by the loss of Cdkn2a and Arf expression, which are both described as critical cell cycle regulators. The loss of Cdkn2a and Arf expression could be mimicked in primary bone marrow-derived mast cells (BMMCs) by sustained SCF supplementation strongly arguing for an involvement of KIT activation in the regulation of Cdkn2a/Arf expression. Hence, this new cell line might be a useful tool to study further aspects of PMC function and to address tumorigenic processes associated with MC leukemia.

Keywords: KIT; Mrgprb2; cell cycle; cyclin-dependent kinase inhibitor; protease; transformation.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

ADP-Ribosylation Factors / metabolism
Animals
Cell Line
Connective Tissue
Cyclin-Dependent Kinase Inhibitor p16 / genetics
Cyclin-Dependent Kinase Inhibitor p16 / metabolism
Humans
Mast Cells*
Mice
Peritoneum*
Proto-Oncogene Proteins c-kit / metabolism
Stem Cell Factor / metabolism

Substances

CDKN2A protein, human
Cdkn2a protein, mouse
Cyclin-Dependent Kinase Inhibitor p16
Stem Cell Factor
Proto-Oncogene Proteins c-kit
ADP-Ribosylation Factors

Grants and funding

This work was supported by grants from the Deutsche Forschungsgemeinschaft (DFG). HU794/12-1 and HU794/14-1 to MH, LI1045/6-1 to CL, WE2554/15-1 to RW and ME3431/2-1 to SM.