Critical role of wnk1 in myc-dependent early mouse thymocyte development

Robert Köchl; Lesley Vanes; Miriam Llorian Sopena; Probir Chakravarty; Harald Hartweger; Kathryn Fountain; Andrea White; Jennifer Cowan; Graham Anderson; Victor L.J. Tybulewicz

doi:10.7554/eLife.56934

Critical role of wnk1 in myc-dependent early mouse thymocyte development

Robert Köchl^*, Lesley Vanes, Miriam Llorian Sopena, Probir Chakravarty, Harald Hartweger, Kathryn Fountain, Andrea White, Jennifer Cowan, Graham Anderson, Victor L.J. Tybulewicz

^*Corresponding author for this work

Immunology and Immunotherapy

Research output: Contribution to journal › Article › peer-review

4 Citations (Scopus)

Abstract

WNK1, a kinase that controls kidney salt homeostasis, also regulates adhesion and migration in CD4⁺ T cells. Wnk1 is highly expressed in thymocytes, and since migration is important for thymocyte maturation, we investigated a role for WNK1 in mouse thymocyte development. We find that WNK1 is required for the transition of double negative (DN) thymocytes through the b-selection checkpoint and subsequent proliferation and differentiation into double positive (DP) thymocytes. Furthermore, we show that WNK1 negatively regulates LFA1-mediated adhesion and positively regulates CXCL12-induced migration in DN thymocytes. Despite this, migration defects of WNK1-deficient thymocytes do not account for the developmental arrest. Instead, we show that in DN thymocytes WNK1 transduces pre-TCR signals via OXSR1 and STK39 kinases, and the SLC12A2 ion co-transporter that are required for post-transcriptional upregulation of MYC and subsequent proliferation and differentiation into DP thymocytes. Thus, a pathway regulating ion homeostasis is a critical regulator of thymocyte development.

Original language	English
Article number	e56934
Pages (from-to)	1-32
Number of pages	32
Journal	eLife
Volume	9
DOIs	https://doi.org/10.7554/eLife.56934
Publication status	Published - Oct 2020

Bibliographical note

Funding Information:
We thank George Kassiotis, Andreas Wack and Edina Schweighoffer for critical reading of the manu-script, Rob de Bruin, Daniel Pennington and Nital Sumaria for advice and Richard Mitter for help with bioinformatics analysis. We thank the Flow Cytometry, Advanced Sequencing, Cell Services and Biological Research Facility of the Francis Crick Institute for flow cytometry, RNA sequencing, provi-sion of cell lines and animal husbandry respectively. We thank Chou-Long Huang, Dario Alessi and Sung-Sen Yang for mouse strains and Warren Pear and Michael Tomasson for retroviral vectors. VLJT was supported by the Biotechnology and Biological Sciences Research Council (grant BB/ L00805X/1), by the UK Medical Research Council (Programme U117527252) and by the Francis Crick Institute which receives its core funding from Cancer Research UK (FC001194), the UK Medical Research Council (FC001194), and the Wellcome Trust (FC001194). GA was supported by the UK Medical Research Council (MR/N000919/1).

Funding Information:
We thank George Kassiotis, Andreas Wack and Edina Schweighoffer for critical reading of the manuscript, Rob de Bruin, Daniel Pennington and Nital Sumaria for advice and Richard Mitter for help with bioinformatics analysis. We thank the Flow Cytometry, Advanced Sequencing, Cell Services and Biological Research Facility of the Francis Crick Institute for flow cytometry, RNA sequencing, provision of cell lines and animal husbandry respectively. We thank Chou-Long Huang, Dario Alessi and Sung-Sen Yang for mouse strains and Warren Pear and Michael Tomasson for retroviral vectors. VLJT was supported by the Biotechnology and Biological Sciences Research Council (grant BB/ L00805X/1), by the UK Medical Research Council (Programme U117527252) and by the Francis Crick Institute which receives its core funding from Cancer Research UK (FC001194), the UK Medical Research Council (FC001194), and the Wellcome Trust (FC001194). GA was supported by the UK Medical Research Council (MR/N000919/1).

Publisher Copyright:
© Köchl et al.

ASJC Scopus subject areas

General Neuroscience
General Biochemistry,Genetics and Molecular Biology
General Immunology and Microbiology

Access to Document

10.7554/eLife.56934

Cite this

@article{9650f129e49e4b1f8204dba4586fdbd1,

title = "Critical role of wnk1 in myc-dependent early mouse thymocyte development",

abstract = "WNK1, a kinase that controls kidney salt homeostasis, also regulates adhesion and migration in CD4+ T cells. Wnk1 is highly expressed in thymocytes, and since migration is important for thymocyte maturation, we investigated a role for WNK1 in mouse thymocyte development. We find that WNK1 is required for the transition of double negative (DN) thymocytes through the b-selection checkpoint and subsequent proliferation and differentiation into double positive (DP) thymocytes. Furthermore, we show that WNK1 negatively regulates LFA1-mediated adhesion and positively regulates CXCL12-induced migration in DN thymocytes. Despite this, migration defects of WNK1-deficient thymocytes do not account for the developmental arrest. Instead, we show that in DN thymocytes WNK1 transduces pre-TCR signals via OXSR1 and STK39 kinases, and the SLC12A2 ion co-transporter that are required for post-transcriptional upregulation of MYC and subsequent proliferation and differentiation into DP thymocytes. Thus, a pathway regulating ion homeostasis is a critical regulator of thymocyte development.",

author = "Robert K{\"o}chl and Lesley Vanes and Sopena, {Miriam Llorian} and Probir Chakravarty and Harald Hartweger and Kathryn Fountain and Andrea White and Jennifer Cowan and Graham Anderson and Tybulewicz, {Victor L.J.}",

note = "Funding Information: We thank George Kassiotis, Andreas Wack and Edina Schweighoffer for critical reading of the manu-script, Rob de Bruin, Daniel Pennington and Nital Sumaria for advice and Richard Mitter for help with bioinformatics analysis. We thank the Flow Cytometry, Advanced Sequencing, Cell Services and Biological Research Facility of the Francis Crick Institute for flow cytometry, RNA sequencing, provi-sion of cell lines and animal husbandry respectively. We thank Chou-Long Huang, Dario Alessi and Sung-Sen Yang for mouse strains and Warren Pear and Michael Tomasson for retroviral vectors. VLJT was supported by the Biotechnology and Biological Sciences Research Council (grant BB/ L00805X/1), by the UK Medical Research Council (Programme U117527252) and by the Francis Crick Institute which receives its core funding from Cancer Research UK (FC001194), the UK Medical Research Council (FC001194), and the Wellcome Trust (FC001194). GA was supported by the UK Medical Research Council (MR/N000919/1). Funding Information: We thank George Kassiotis, Andreas Wack and Edina Schweighoffer for critical reading of the manuscript, Rob de Bruin, Daniel Pennington and Nital Sumaria for advice and Richard Mitter for help with bioinformatics analysis. We thank the Flow Cytometry, Advanced Sequencing, Cell Services and Biological Research Facility of the Francis Crick Institute for flow cytometry, RNA sequencing, provision of cell lines and animal husbandry respectively. We thank Chou-Long Huang, Dario Alessi and Sung-Sen Yang for mouse strains and Warren Pear and Michael Tomasson for retroviral vectors. VLJT was supported by the Biotechnology and Biological Sciences Research Council (grant BB/ L00805X/1), by the UK Medical Research Council (Programme U117527252) and by the Francis Crick Institute which receives its core funding from Cancer Research UK (FC001194), the UK Medical Research Council (FC001194), and the Wellcome Trust (FC001194). GA was supported by the UK Medical Research Council (MR/N000919/1). Publisher Copyright: {\textcopyright} K{\"o}chl et al.",

year = "2020",

month = oct,

doi = "10.7554/eLife.56934",

language = "English",

volume = "9",

pages = "1--32",

journal = "eLife",

issn = "2050-084X",

publisher = "eLife Sciences Publications",

}

TY - JOUR

T1 - Critical role of wnk1 in myc-dependent early mouse thymocyte development

AU - Köchl, Robert

AU - Vanes, Lesley

AU - Sopena, Miriam Llorian

AU - Chakravarty, Probir

AU - Hartweger, Harald

AU - Fountain, Kathryn

AU - White, Andrea

AU - Cowan, Jennifer

AU - Anderson, Graham

AU - Tybulewicz, Victor L.J.

N1 - Funding Information: We thank George Kassiotis, Andreas Wack and Edina Schweighoffer for critical reading of the manu-script, Rob de Bruin, Daniel Pennington and Nital Sumaria for advice and Richard Mitter for help with bioinformatics analysis. We thank the Flow Cytometry, Advanced Sequencing, Cell Services and Biological Research Facility of the Francis Crick Institute for flow cytometry, RNA sequencing, provi-sion of cell lines and animal husbandry respectively. We thank Chou-Long Huang, Dario Alessi and Sung-Sen Yang for mouse strains and Warren Pear and Michael Tomasson for retroviral vectors. VLJT was supported by the Biotechnology and Biological Sciences Research Council (grant BB/ L00805X/1), by the UK Medical Research Council (Programme U117527252) and by the Francis Crick Institute which receives its core funding from Cancer Research UK (FC001194), the UK Medical Research Council (FC001194), and the Wellcome Trust (FC001194). GA was supported by the UK Medical Research Council (MR/N000919/1). Funding Information: We thank George Kassiotis, Andreas Wack and Edina Schweighoffer for critical reading of the manuscript, Rob de Bruin, Daniel Pennington and Nital Sumaria for advice and Richard Mitter for help with bioinformatics analysis. We thank the Flow Cytometry, Advanced Sequencing, Cell Services and Biological Research Facility of the Francis Crick Institute for flow cytometry, RNA sequencing, provision of cell lines and animal husbandry respectively. We thank Chou-Long Huang, Dario Alessi and Sung-Sen Yang for mouse strains and Warren Pear and Michael Tomasson for retroviral vectors. VLJT was supported by the Biotechnology and Biological Sciences Research Council (grant BB/ L00805X/1), by the UK Medical Research Council (Programme U117527252) and by the Francis Crick Institute which receives its core funding from Cancer Research UK (FC001194), the UK Medical Research Council (FC001194), and the Wellcome Trust (FC001194). GA was supported by the UK Medical Research Council (MR/N000919/1). Publisher Copyright: © Köchl et al.

PY - 2020/10

Y1 - 2020/10

N2 - WNK1, a kinase that controls kidney salt homeostasis, also regulates adhesion and migration in CD4+ T cells. Wnk1 is highly expressed in thymocytes, and since migration is important for thymocyte maturation, we investigated a role for WNK1 in mouse thymocyte development. We find that WNK1 is required for the transition of double negative (DN) thymocytes through the b-selection checkpoint and subsequent proliferation and differentiation into double positive (DP) thymocytes. Furthermore, we show that WNK1 negatively regulates LFA1-mediated adhesion and positively regulates CXCL12-induced migration in DN thymocytes. Despite this, migration defects of WNK1-deficient thymocytes do not account for the developmental arrest. Instead, we show that in DN thymocytes WNK1 transduces pre-TCR signals via OXSR1 and STK39 kinases, and the SLC12A2 ion co-transporter that are required for post-transcriptional upregulation of MYC and subsequent proliferation and differentiation into DP thymocytes. Thus, a pathway regulating ion homeostasis is a critical regulator of thymocyte development.

AB - WNK1, a kinase that controls kidney salt homeostasis, also regulates adhesion and migration in CD4+ T cells. Wnk1 is highly expressed in thymocytes, and since migration is important for thymocyte maturation, we investigated a role for WNK1 in mouse thymocyte development. We find that WNK1 is required for the transition of double negative (DN) thymocytes through the b-selection checkpoint and subsequent proliferation and differentiation into double positive (DP) thymocytes. Furthermore, we show that WNK1 negatively regulates LFA1-mediated adhesion and positively regulates CXCL12-induced migration in DN thymocytes. Despite this, migration defects of WNK1-deficient thymocytes do not account for the developmental arrest. Instead, we show that in DN thymocytes WNK1 transduces pre-TCR signals via OXSR1 and STK39 kinases, and the SLC12A2 ion co-transporter that are required for post-transcriptional upregulation of MYC and subsequent proliferation and differentiation into DP thymocytes. Thus, a pathway regulating ion homeostasis is a critical regulator of thymocyte development.

UR - http://www.scopus.com/inward/record.url?scp=85094932787&partnerID=8YFLogxK

U2 - 10.7554/eLife.56934

DO - 10.7554/eLife.56934

M3 - Article

C2 - 33051000

AN - SCOPUS:85094932787

SN - 2050-084X

VL - 9

SP - 1

EP - 32

JO - eLife

JF - eLife

M1 - e56934

ER -

Critical role of wnk1 in myc-dependent early mouse thymocyte development

Abstract

Bibliographical note

ASJC Scopus subject areas

Access to Document

Fingerprint

Cite this