PTPN22 R620W gene editing in T cells enhances low avidity TCR responses
- Seattle Children's Research Institute, United States
- Benaroya Research Institute, United States
- University of Washington, United States
Abstract
A genetic variant in the gene PTPN22 (R620W, rs2476601) is strongly associated with increased risk for multiple autoimmune diseases and linked to altered TCR regulation and T cell activation. Here, we utilize Crispr/Cas9 gene editing with donor DNA repair templates in human cord blood-derived, naive T cells to generate PTPN22 risk edited (620W), non-risk edited (620R) or knock out T cells from the same donor. PTPN22 risk edited cells exhibited increased activation marker expression following non-specific TCR engagement, findings that mimicked PTPN22 KO cells. Next, using lentiviral delivery of T1D patient-derived TCRs against the pancreatic autoantigen, islet-specific glucose-6 phosphatase catalytic subunit-related protein (IGRP), we demonstrate that loss of PTPN22 function led to enhanced signaling in T cells expressing a lower avidity self-reactive TCR, but not a high avidity TCR. In this setting, loss of PTPN22 mediated enhanced proliferation and Th1 skewing. Importantly, expression of the risk variant in association with a lower avidity TCR also increased proliferation relative to PTPN22 non-risk T cells. Together, these findings suggest that, in primary human T cells, PTPN22 rs2476601 contributes to autoimmunity risk by permitting increased TCR signaling and activation in mildly self-reactive T cells, thereby potentially expanding the self-reactive T cell pool and skewing this population toward an inflammatory phenotype.
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Prism files containing the numerical data and statistical tests used to generate all figures has been provided
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Author details
Funding
National Institute of Diabetes and Digestive and Kidney Diseases (DP3DK111802)
- Warren Anderson
- Fariba Barahmand-pour-Whitman
- Peter S Linsley
- Karen Cerosaletti
- Jane H Buckner
- David J Rawlings
Seattle Children's Foundation (Children's Guild Association Endowed Chair in Pediatric Immunology)
- David J Rawlings
Seattle Children's Research Institute (Center for Immunity and Immunotherapies)
- David J Rawlings
Seattle Children's Research Institute (Program for Cell and Gene Therapy)
- David J Rawlings
Seattle Children's Research Institute (Hansen Investigator in Pediatric Innovation Endowment)
- David J Rawlings
Benaroya Family Gift Fund
- David J Rawlings
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Ethics
Human subjects: Study Approval (Human Subjects)For gene editing experiments using adult PBMCs, human donor leukopaks were purchased from the Fred Hutchinson Cancer Research Center, which were obtained from consenting donors under an IRB-approved protocol and cryopreserved. For gene editing experiments using umbilical cord blood derived PBMCs, cord units were purchased from the Bloodworks Northwest, which were obtained with consent under an IRB-approved protocol and cryopreserved. After collection, all samples were de-identified for the protection of human blood donors.
Copyright
© 2023, Anderson et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
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PTPN22 R620W gene editing in T cells enhances low avidity TCR responses
eLife 12:e81577.
https://doi.org/10.7554/eLife.81577
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