CRISPR-Cas9-Mediated Glucocorticoid Resistance in Virus-Specific T Cells for Adoptive T Cell Therapy Posttransplantation.

Abstract

Immunosuppression posttransplantation exposes patients to an increased risk for refractory viral infections as an important cause of morbidity and mortality. Protective T cell immunity can be restored by adoptive T cell transfer, but ongoing immunosuppression limits efficacy of T cell responses. In order to deliver protection against viral pathogens and allow at the same time necessary steroid therapy, we generated glucocorticoid-resistant T cells by CRISPR-Cas9-mediated knockout of the glucocorticoid receptor in primary human virus-specific T cell products. Characterization of the T cell product revealed high efficiency of glucocorticoid receptor knockout and high purity of virus-specific T cells. This tandem T cell engineering preserved protective T cell functionality, such as cytotoxicity, CD107a degranulation, proliferative capacity, and cytokine release patterns. Virus-specific T cells with glucocorticoid receptor knockout were resistant to the suppressive effect of dexamethasone treatment on lymphocyte proliferation and cytokine secretion (tumor necrosis factor alpha [TNF-α], interleukin-4 [IL-4], IL-6, and sFas). Additionally, glucocorticoid receptor knockout cells remained sensitive to cyclosporine A treatment, thereby providing a rescue approach for patients in case of safety issues. This novel approach provides a therapeutic option for the treatment of patients with viral infections after transplantation who are receiving glucocorticoid therapy.

• Authors: Theresa Kaeuferle, Larissa Deisenberger, Lena Jablonowski, Tanja A Stief, Franziska Blaeschke, Semjon Willier, Tobias Feuchtinger
• Journal: Molecular therapy : the journal of the American Society of Gene Therapy (Mol Ther) Vol. 28 Issue 9 Pg. 1965-1973 (09 02 2020) ISSN: 1525-0024 [Electronic] United States
• PMID: 32559432 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2020 The American Society of Gene and Cell Therapy. Published by Elsevier Inc. All rights reserved.

Chemical References

• Cytokines
• Glucocorticoids
• Receptors, Glucocorticoid
• Viral Matrix Proteins
• cytomegalovirus matrix protein 65kDa
• Cyclosporine

Topics

• Adoptive Transfer (methods)
• CRISPR-Cas Systems
• Cell Engineering (methods)
• Cell Proliferation (genetics)
• Cells, Cultured
• Cyclosporine (pharmacology)
• Cytokines (metabolism)
• Drug Resistance (genetics)
• Gene Knockout Techniques (methods)
• Glucocorticoids (therapeutic use)
• Hematopoietic Stem Cell Transplantation (adverse effects)
• Humans
• Lymphocyte Activation (immunology)
• Receptors, Glucocorticoid (deficiency, genetics)
• T-Lymphocytes (drug effects, immunology, metabolism)
• Viral Matrix Proteins (immunology)
• Virus Diseases (etiology, immunology, therapy)