A novel knockout mouse model to assess the impact of one-copy loss of Hnrnpk in CD4 + T cells in chronically inflamed skin as a prelude to CTCL

Abstract Cutaneous T-cell lymphomas (CTCLs), particularly Mycosis fungoides (MF), frequently exhibit deletions and reduced expression of HNRNPK in CD4 + T cells. To enable in vivo studies, we developed a conditional Hnrnpk knockout mouse that thrives, facilitating the investigation of HNRNPK’s role...

Full description

Saved in:
Bibliographic Details
Main Authors: Yixin Luo, Maarten Vermeer, Margot M. Linssen, Fenna J. de Bie, Marloe Pijnacker-Verspuij, Conny Brouwers, Jill Claassens, Frank R. de Gruijl, Peter Hohenstein, Cornelis P. Tensen
Format: Article
Language:English
Published: Nature Portfolio 2025-04-01
Series:Scientific Reports
Subjects:
Online Access:https://doi.org/10.1038/s41598-025-98640-6
Tags: Add Tag
No Tags, Be the first to tag this record!
_version_ 1849713568358858752
author Yixin Luo
Maarten Vermeer
Margot M. Linssen
Fenna J. de Bie
Marloe Pijnacker-Verspuij
Conny Brouwers
Jill Claassens
Frank R. de Gruijl
Peter Hohenstein
Cornelis P. Tensen
author_facet Yixin Luo
Maarten Vermeer
Margot M. Linssen
Fenna J. de Bie
Marloe Pijnacker-Verspuij
Conny Brouwers
Jill Claassens
Frank R. de Gruijl
Peter Hohenstein
Cornelis P. Tensen
author_sort Yixin Luo
collection DOAJ
description Abstract Cutaneous T-cell lymphomas (CTCLs), particularly Mycosis fungoides (MF), frequently exhibit deletions and reduced expression of HNRNPK in CD4 + T cells. To enable in vivo studies, we developed a conditional Hnrnpk knockout mouse that thrives, facilitating the investigation of HNRNPK’s role in CTCL onset. We generated mice with a floxed Hnrnpk allele, then crossbred them with Cd4CreER T 2 mice to generate Hnrnpk flox Cd4CreER T 2 mice, all in BL6 background. PCR confirmed the targeted deletion of Hnrnpk in CD4 + T cells after tamoxifen i.p. injection. Skin allergic reactions were induced with oxazolone, and Cre was activated in skin-infiltrating CD4 + T cells using tamoxifen topically after the first allergic skin reaction. The mice exhibited no immediately obvious phenotype. Flow cytometry and histopathological analysis were conducted on blood and skin samples collected throughout the experiment. Following 20 weeks of sustained allergic reactions, inflammation persisted over 20 weeks after challenges ceased, demonstrating early CTCL characteristics such as chronic skin inflammation, CD3 + CD4 + T cell infiltration, and stable peripheral blood parameters. This mouse model provides experimental access to the complex microenvironment and immune responses involved in early inflammatory stages, providing opportunities for further research into the role of HNRNPK in CTCL and the development of effective therapeutic interventions for this challenging malignancy.
format Article
id doaj-art-df512cd63be4446eaefb3de66cf5b3ae
institution DOAJ
issn 2045-2322
language English
publishDate 2025-04-01
publisher Nature Portfolio
record_format Article
series Scientific Reports
spelling doaj-art-df512cd63be4446eaefb3de66cf5b3ae2025-08-20T03:13:55ZengNature PortfolioScientific Reports2045-23222025-04-0115111110.1038/s41598-025-98640-6A novel knockout mouse model to assess the impact of one-copy loss of Hnrnpk in CD4 + T cells in chronically inflamed skin as a prelude to CTCLYixin Luo0Maarten Vermeer1Margot M. Linssen2Fenna J. de Bie3Marloe Pijnacker-Verspuij4Conny Brouwers5Jill Claassens6Frank R. de Gruijl7Peter Hohenstein8Cornelis P. Tensen9Department of Dermatology, Leiden University Medical CenterDepartment of Dermatology, Leiden University Medical CenterTransgenic Facility Leiden, Central Animal Facility, Leiden University Medical CenterDepartment of Dermatology, Leiden University Medical CenterExperimental Animal Facility, Leiden University Medical CenterTransgenic Facility Leiden, Central Animal Facility, Leiden University Medical CenterTransgenic Facility Leiden, Central Animal Facility, Leiden University Medical CenterDepartment of Dermatology, Leiden University Medical CenterTransgenic Facility Leiden, Central Animal Facility, Leiden University Medical CenterDepartment of Dermatology, Leiden University Medical CenterAbstract Cutaneous T-cell lymphomas (CTCLs), particularly Mycosis fungoides (MF), frequently exhibit deletions and reduced expression of HNRNPK in CD4 + T cells. To enable in vivo studies, we developed a conditional Hnrnpk knockout mouse that thrives, facilitating the investigation of HNRNPK’s role in CTCL onset. We generated mice with a floxed Hnrnpk allele, then crossbred them with Cd4CreER T 2 mice to generate Hnrnpk flox Cd4CreER T 2 mice, all in BL6 background. PCR confirmed the targeted deletion of Hnrnpk in CD4 + T cells after tamoxifen i.p. injection. Skin allergic reactions were induced with oxazolone, and Cre was activated in skin-infiltrating CD4 + T cells using tamoxifen topically after the first allergic skin reaction. The mice exhibited no immediately obvious phenotype. Flow cytometry and histopathological analysis were conducted on blood and skin samples collected throughout the experiment. Following 20 weeks of sustained allergic reactions, inflammation persisted over 20 weeks after challenges ceased, demonstrating early CTCL characteristics such as chronic skin inflammation, CD3 + CD4 + T cell infiltration, and stable peripheral blood parameters. This mouse model provides experimental access to the complex microenvironment and immune responses involved in early inflammatory stages, providing opportunities for further research into the role of HNRNPK in CTCL and the development of effective therapeutic interventions for this challenging malignancy.https://doi.org/10.1038/s41598-025-98640-6Cutaneous T-cell lymphomasMycosis fungoidesCD4 + T cellsHNRNPKTransgenic mouse
spellingShingle Yixin Luo
Maarten Vermeer
Margot M. Linssen
Fenna J. de Bie
Marloe Pijnacker-Verspuij
Conny Brouwers
Jill Claassens
Frank R. de Gruijl
Peter Hohenstein
Cornelis P. Tensen
A novel knockout mouse model to assess the impact of one-copy loss of Hnrnpk in CD4 + T cells in chronically inflamed skin as a prelude to CTCL
Scientific Reports
Cutaneous T-cell lymphomas
Mycosis fungoides
CD4 + T cells
HNRNPK
Transgenic mouse
title A novel knockout mouse model to assess the impact of one-copy loss of Hnrnpk in CD4 + T cells in chronically inflamed skin as a prelude to CTCL
title_full A novel knockout mouse model to assess the impact of one-copy loss of Hnrnpk in CD4 + T cells in chronically inflamed skin as a prelude to CTCL
title_fullStr A novel knockout mouse model to assess the impact of one-copy loss of Hnrnpk in CD4 + T cells in chronically inflamed skin as a prelude to CTCL
title_full_unstemmed A novel knockout mouse model to assess the impact of one-copy loss of Hnrnpk in CD4 + T cells in chronically inflamed skin as a prelude to CTCL
title_short A novel knockout mouse model to assess the impact of one-copy loss of Hnrnpk in CD4 + T cells in chronically inflamed skin as a prelude to CTCL
title_sort novel knockout mouse model to assess the impact of one copy loss of hnrnpk in cd4 t cells in chronically inflamed skin as a prelude to ctcl
topic Cutaneous T-cell lymphomas
Mycosis fungoides
CD4 + T cells
HNRNPK
Transgenic mouse
url https://doi.org/10.1038/s41598-025-98640-6
work_keys_str_mv AT yixinluo anovelknockoutmousemodeltoassesstheimpactofonecopylossofhnrnpkincd4tcellsinchronicallyinflamedskinasapreludetoctcl
AT maartenvermeer anovelknockoutmousemodeltoassesstheimpactofonecopylossofhnrnpkincd4tcellsinchronicallyinflamedskinasapreludetoctcl
AT margotmlinssen anovelknockoutmousemodeltoassesstheimpactofonecopylossofhnrnpkincd4tcellsinchronicallyinflamedskinasapreludetoctcl
AT fennajdebie anovelknockoutmousemodeltoassesstheimpactofonecopylossofhnrnpkincd4tcellsinchronicallyinflamedskinasapreludetoctcl
AT marloepijnackerverspuij anovelknockoutmousemodeltoassesstheimpactofonecopylossofhnrnpkincd4tcellsinchronicallyinflamedskinasapreludetoctcl
AT connybrouwers anovelknockoutmousemodeltoassesstheimpactofonecopylossofhnrnpkincd4tcellsinchronicallyinflamedskinasapreludetoctcl
AT jillclaassens anovelknockoutmousemodeltoassesstheimpactofonecopylossofhnrnpkincd4tcellsinchronicallyinflamedskinasapreludetoctcl
AT frankrdegruijl anovelknockoutmousemodeltoassesstheimpactofonecopylossofhnrnpkincd4tcellsinchronicallyinflamedskinasapreludetoctcl
AT peterhohenstein anovelknockoutmousemodeltoassesstheimpactofonecopylossofhnrnpkincd4tcellsinchronicallyinflamedskinasapreludetoctcl
AT cornelisptensen anovelknockoutmousemodeltoassesstheimpactofonecopylossofhnrnpkincd4tcellsinchronicallyinflamedskinasapreludetoctcl
AT yixinluo novelknockoutmousemodeltoassesstheimpactofonecopylossofhnrnpkincd4tcellsinchronicallyinflamedskinasapreludetoctcl
AT maartenvermeer novelknockoutmousemodeltoassesstheimpactofonecopylossofhnrnpkincd4tcellsinchronicallyinflamedskinasapreludetoctcl
AT margotmlinssen novelknockoutmousemodeltoassesstheimpactofonecopylossofhnrnpkincd4tcellsinchronicallyinflamedskinasapreludetoctcl
AT fennajdebie novelknockoutmousemodeltoassesstheimpactofonecopylossofhnrnpkincd4tcellsinchronicallyinflamedskinasapreludetoctcl
AT marloepijnackerverspuij novelknockoutmousemodeltoassesstheimpactofonecopylossofhnrnpkincd4tcellsinchronicallyinflamedskinasapreludetoctcl
AT connybrouwers novelknockoutmousemodeltoassesstheimpactofonecopylossofhnrnpkincd4tcellsinchronicallyinflamedskinasapreludetoctcl
AT jillclaassens novelknockoutmousemodeltoassesstheimpactofonecopylossofhnrnpkincd4tcellsinchronicallyinflamedskinasapreludetoctcl
AT frankrdegruijl novelknockoutmousemodeltoassesstheimpactofonecopylossofhnrnpkincd4tcellsinchronicallyinflamedskinasapreludetoctcl
AT peterhohenstein novelknockoutmousemodeltoassesstheimpactofonecopylossofhnrnpkincd4tcellsinchronicallyinflamedskinasapreludetoctcl
AT cornelisptensen novelknockoutmousemodeltoassesstheimpactofonecopylossofhnrnpkincd4tcellsinchronicallyinflamedskinasapreludetoctcl