title: Blocking TCR restimulation induced necroptosis in adoptively transferred T cells improves tumor control.
abstract: Advancements in adoptive cell transfer therapy (ACT) has led to the use of T cells engineered with tumor specific T cell receptors, which after rapid expansion can be obtained in sufficient numbers for treating patients. However, due to massive proliferation these cells are close to replicative senescence, exhibit exhausted phenotype, and also display increased susceptibility to activation induced cell death. We have previously shown that tumor reactive T cells undergo caspase-independent cell death upon TCR restimulation with cognate antigen, which involves reactive oxygen species and c-jun N-terminal kinase. Herein, we show that a large fraction of the human melanoma epitope tyrosinase reactive TCR transduced T cells that exhibit effector memory (TEM) phenotype and undergo programmed necrosis, or necroptosis, upon TCR restimulation. As compared to the T central memory (TCM) subsets, the TEM subset displayed an increased expression of genes involved in necroptotic cell death, and a necrotic phenotype upon TCR restimulation as confirmed by electron microscopy. Higher expression of receptor-interacting kinases (RIPK) that mediate necroptosis was also observed in the TEM fraction. Further, the TEM cells were rescued from undergoing necroptosis when pretreated with necroptotic inhibitor NecroX2 before TCR restimulation. Importantly, NecroX2 pretreated tumor reactive T cells also exhibited better tumor control and increased in vivo persistence when adoptively-transferred to treat subcutaneously established murine melanoma B16-F10. Thus, it is likely that the outcome of ACT could be vastly improved by interfering with the necroptotic cell death pathway in activated tumor reactive T cells used in immunotherapy.
authors.affiliation: Department of Surgery, Medical University of South Carolina, Charleston, SC, USA.
authors.forename: Pravin
authors.initials: P
authors.lastname: Kesarwani
authors.affiliation: Department of Surgery, Medical University of South Carolina, Charleston, SC, USA.
authors.forename: Paramita
authors.initials: P
authors.lastname: Chakraborty
authors.affiliation: Department of Microbiology & Immunology, Medical University of South Carolina, Charleston, SC, USA.
authors.forename: Radhika
authors.initials: R
authors.lastname: Gudi
authors.affiliation: Department of Surgery, Medical University of South Carolina, Charleston, SC, USA.
authors.forename: Shilpak
authors.initials: S
authors.lastname: Chatterjee
authors.affiliation: Department of Surgery, Loyola University, Maywood, IL, USA.
authors.forename: Gina
authors.initials: G
authors.lastname: Scurti
authors.affiliation: Department of Surgery, Medical University of South Carolina, Charleston, SC, USA.
authors.forename: Kyle
authors.initials: K
authors.lastname: Toth
authors.affiliation: Department of Surgery, Loyola University, Maywood, IL, USA.
authors.forename: Patt
authors.initials: P
authors.lastname: Simms
authors.affiliation: Department of Surgery, Medical University of South Carolina, Charleston, SC, USA.
authors.forename: Mahvash
authors.initials: M
authors.lastname: Husain
authors.affiliation: Department of Public Health, Medical University of South Carolina, Charleston, SC, USA.
authors.forename: Kent
authors.initials: K
authors.lastname: Armeson
authors.affiliation: Department of Opthamology, Medical University of South Carolina, Charleston, SC, USA.
authors.forename: Shahid
authors.initials: S
authors.lastname: Husain
authors.affiliation: Department of Public Health, Medical University of South Carolina, Charleston, SC, USA.
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Garrett-Mayer
authors.affiliation: Department of Microbiology & Immunology, Medical University of South Carolina, Charleston, SC, USA.
authors.forename: Chethamarakshan
authors.initials: C
authors.lastname: Vasu
authors.affiliation: Department of Surgery, Loyola University, Maywood, IL, USA.
authors.forename: Michael I
authors.initials: MI
authors.lastname: Nishimura
authors.affiliation: Department of Surgery, Medical University of South Carolina, Charleston, SC, USA.
authors.forename: Shikhar
authors.initials: S
authors.lastname: Mehrotra
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000276:immunology*; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology / Q000378:metabolism / Q000637:transplantation*; D045744:Cell Line, Tumor; D006801:Humans; D016219:Immunotherapy, Adoptive / Q000379:methods*; D007371:Interferon-gamma / Q000276:immunology / Q000378:metabolism; D019041:L-Selectin / Q000276:immunology / Q000378:metabolism; D008546:Melanoma, Experimental / Q000276:immunology / Q000378:metabolism / Q000628:therapy*; D008810:Mice, Inbred C57BL; D009336:Necrosis / Q000276:immunology; D053422:Receptor-Interacting Protein Serine-Threonine Kinases / Q000276:immunology / Q000378:metabolism; D011948:Receptors, Antigen, T-Cell / Q000235:genetics / Q000276:immunology* / Q000378:metabolism
chemical_list: D011948:Receptors, Antigen, T-Cell; D019041:L-Selectin; D007371:Interferon-gamma; C506249:RIPK1 protein, human; C119705:RIPK3 protein, human; D053422:Receptor-Interacting Protein Serine-Threonine Kinases
keywords: Immune response; Immunity; Immunology and Microbiology Section; T cell; apoptosis; immunotherapy; melanoma