title: Noninvasive detection of tumor-infiltrating T cells by PET reporter imaging.
abstract: Adoptive transfer of tumor-reactive T cells can successfully reduce tumor burden; however, in rare cases, lethal on-target/off-tumor effects have been reported. A noninvasive method to track engineered cells with high sensitivity and resolution would allow observation of correct cell homing and/or identification of dangerous off-target locations in preclinical and clinical applications. Human deoxycytidine kinase triple mutant (hdCK3mut) is a nonimmunogenic PET reporter that was previously shown to be an effective tool to monitor whole-body hematopoiesis. Here, we engineered a construct in which hdCK3mut is coexpressed with the anti-melanoma T cell receptor F5, introduced this construct into human CD34 cells or PBMCs, and evaluated this approach in multiple immunotherapy models. Expression of hdCK3mut allowed engrafted cells to be visualized within recipient bone marrow, while accumulation of [18F]-L-FMAU in hdCK3mut-expressing T cells permitted detection of intratumoral homing. Animals that received T cells coexpressing hdCK3mut and the anti-melanoma T cell receptor had demonstrably higher signals in HLA-matched tumors compared with those in animals that received cells solely expressing hdCK3mut. Engineered T cells caused cytotoxicity in HLA/antigen-matched tumors and induced IFN-γ production and activation. Moreover, hdCK3mut permitted simultaneous monitoring of engraftment and tumor infiltration, without affecting T cell function. Our findings suggest that hdCK3mut reporter imaging can be applied in clinical immunotherapies for whole-body detection of engineered cell locations.
authors.forename: Melissa N
authors.initials: MN
authors.lastname: McCracken
authors.forename: Dimitrios N
authors.initials: DN
authors.lastname: Vatakis
authors.forename: Dhaval
authors.initials: D
authors.lastname: Dixit
authors.forename: Jami
authors.initials: J
authors.lastname: McLaughlin
authors.forename: Jerome A
authors.initials: JA
authors.lastname: Zack
authors.forename: Owen N
authors.initials: ON
authors.lastname: Witte
mesh_terms: D000818:Animals; D001853:Bone Marrow / Q000000981:diagnostic imaging; D002634:Chemotaxis, Leukocyte; D003601:Cytotoxicity Tests, Immunologic; D003842:Deoxycytidine Kinase / Q000032:analysis* / Q000235:genetics; D017930:Genes, Reporter; D005813:Genes, Synthetic; D005822:Genetic Vectors / Q000235:genetics; D006085:Graft Survival; D015789:HLA-A2 Antigen / Q000276:immunology; D018380:Hematopoietic Stem Cell Transplantation; D006412:Hematopoietic Stem Cells / Q000737:chemistry / Q000502:physiology; D006801:Humans; D007167:Immunotherapy / Q000379:methods*; D016219:Immunotherapy, Adoptive; D059425:Interferon-gamma Release Tests; D016086:Lentivirus / Q000235:genetics; D007963:Leukocytes, Mononuclear / Q000737:chemistry / Q000502:physiology / Q000637:transplantation; D016246:Lymphocytes, Tumor-Infiltrating / Q000276:immunology*; D058965:MART-1 Antigen / Q000276:immunology; D008546:Melanoma, Experimental / Q000276:immunology / Q000628:therapy*; D051379:Mice; D009154:Mutation; D049268:Positron-Emission Tomography; D011948:Receptors, Antigen, T-Cell / Q000032:analysis / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000235:genetics; D012190:Retroviridae / Q000235:genetics; D013601:T-Lymphocytes / Q000276:immunology* / Q000637:transplantation; D013950:Thymus Gland / Q000637:transplantation
chemical_list: D015789:HLA-A2 Antigen; D058965:MART-1 Antigen; D011948:Receptors, Antigen, T-Cell; D011993:Recombinant Fusion Proteins; D003842:Deoxycytidine Kinase
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