eprintid: 10203183
rev_number: 14
eprint_status: archive
userid: 699
dir: disk0/10/20/31/83
datestamp: 2025-02-28 07:53:03
lastmod: 2025-02-28 07:53:03
status_changed: 2025-02-28 07:53:03
type: thesis
metadata_visibility: show
sword_depositor: 699
creators_name: Joseph, Megan Daisy
title: Illuminating the role of the protein tyrosine phosphatase PTPN22 in actin remodelling by super-resolution microscopy: insights into T cell synapse formation
ispublished: unpub
divisions: UCL
divisions: B02
divisions: C08
divisions: D09
note: Copyright © The Author 2025.   Original content in this thesis is licensed under the terms of the Creative Commons Attribution-NonCommercial 4.0 International (CC BY-NC 4.0) Licence (https://creativecommons.org/licenses/by-nc/4.0/).  Any third-party copyright material present remains the property of its respective owner(s) and is licensed under its existing terms.  Access may initially be restricted at the author’s request.
abstract: Protein Tyrosine Phosphatase Non-Receptor Type 22 (PTPN22) is a protein tyrosine phosphatase
operating within the T cell signalling cascade. PTPN22 dephosphorylates key signalling proteins,
in combination with the binding partner c-SRC kinase (Csk), which leads to downregulation of T
cell activation. Associated with over 15 autoimmune diseases, PTPN22 R620W is one of the most
common autoimmune disease related mutations. Recently, studies have uncovered a novel binding
partner of PTPN22, proline-serine-threonine phosphatase interacting protein 1 (PSTPIP-1). Whilst
decades of research have focused on understanding the consequences of PTPN22 mutation in T cell
activation, the effects of the PTPN22-PSTPIP-1 interaction are poorly understood. PSTPIP-1 regulates F-actin remodelling via association with Wiskott-Aldrich syndrome protein (WASp). WASp
promotes the formation of F-actin foci following T cell receptor (TCR) microcluster formation,
amplifying downstream TCR signals. Mutations within PSTPIP-1 are associated with autoinflammatory disorders and immunodeficiencies. These mutations correlate with defects in F-actin
formation and lie within the PTPN22 binding domain, suggesting that PTPN22/PSTPIP-1 interactions regulate actin remodelling in T cells and thus T cell signalling. Using both live and fixed
multicolour 3D super-resolution imaging on a Spinning Disk Super Resolution by Optical Pixel
Reassignment (SoRa) microscope, this thesis describes a series of experiments exploiting Jurkat T
cells deficient in PTPN22 expression and expressing T cell receptors reactive to ligands of different
affinity. A key finding is that Jurkat T cells lacking PTPN22 exhibit aberrant actin remodelling
along with disrupted PSTPIP-1 and TCR clustering upon activation. The nanoscale organisation
of PSTPIP-1 and TCR clusters at different Z-planes was visualised using single molecule localization DNA-PAINT microscopy on the SoRa microscope, coupled with quantitative PAINT analysis.
These results uncover a novel pathway involving PTPN22 and PSTPIP-1 in T cell signalling, providing new insight into the molecular mechanisms that underpin susceptibility to a wide range of
autoimmune diseases.
date: 2025-01-28
date_type: published
oa_status: green
full_text_type: other
thesis_class: doctoral_open
thesis_award: Ph.D
language: eng
primo: open
primo_central: open_green
verified: verified_manual
elements_id: 2351976
lyricists_name: Joseph, Megan
lyricists_id: MDJOS56
actors_name: Joseph, Megan
actors_id: MDJOS56
actors_role: owner
full_text_status: public
pagerange: 1-1
pages: 263
institution: UCL (University College London)
department: London Centre for Nanotechnology
thesis_type: Doctoral
citation:        Joseph, Megan Daisy;      (2025)    Illuminating the role of the protein tyrosine phosphatase PTPN22 in actin remodelling by super-resolution microscopy: insights into T cell synapse formation.                   Doctoral thesis  (Ph.D), UCL (University College London).     Green open access   
 
document_url: https://discovery.ucl.ac.uk/id/eprint/10203183/1/Joseph-10203183_thesis.pdf