CRISPR-Cas9-Edited Tacrolimus-Resistant Antiviral T Cells for Advanced Adoptive Immunotherapy in Transplant Recipients.
Viral infections, such as with cytomegalovirus (CMV), remain a major risk factor for mortality and morbidity of transplant recipients because of their requirement for lifelong immunosuppression (IS). Antiviral drugs often cause toxicity and sometimes fail to control disease. Thus, regeneration of the antiviral immune response by adoptive antiviral T cell therapy is an attractive alternative. Our recent data, however, show only short-term efficacy in some solid organ recipients, possibly because of malfunction in transferred T cells caused by ongoing IS. We developed a vector-free clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9-based good manufacturing practice (GMP)-compliant protocol that efficiently targets and knocks out the gene for the adaptor protein FK506-binding protein 12 (FKBP12), required for the immunosuppressive function of tacrolimus. This was achieved by transient delivery of ribonucleoprotein complexes into CMV-specific T cells by electroporation. We confirmed the tacrolimus resistance of our gene-edited T cell products in vitro and demonstrated performance comparable with non-tacrolimus-treated unmodified T cells. The alternative calcineurin inhibitor cyclosporine A can be administered as a safety switch to shut down tacrolimus-resistant T cell activity in case of adverse effects. Furthermore, we performed safety assessments as a prerequisite for translation to first-in-human applications.
['Amini|Leila|L|', 'Wagner|Dimitrios Laurin|DL|', 'Rössler|Uta|U|', 'Zarrinrad|Ghazaleh|G|', 'Wagner|Livia Felicitas|LF|', 'Vollmer|Tino|T|', 'Wendering|Désirée Jacqueline|DJ|', 'Kornak|Uwe|U|', 'Volk|Hans-Dieter|HD|', 'Reinke|Petra|P|', 'Schmueck-Henneresse|Michael|M|']
D064113:CRISPR-Cas Systems D060467:Disease Resistance D004351:Drug Resistance D000072669:Gene Editing D005818:Genetic Engineering D006801:Humans D007166:Immunosuppressive Agents D016219:Immunotherapy, Adoptive D013601:T-Lymphocytes D016559:Tacrolimus D066027:Transplant Recipients
immunotherapy CMV immunosuppression CRISPR-Cas9 adoptive T-cell-therapy drug-resistance solid-organ-transplantation vector-free gene-editing