Novel T cells with improved in vivo anti-tumor activity generated by RNA electroporation
Received date: 16 Feb 2017
Accepted date: 20 Apr 2017
Published date: 23 Aug 2017
Copyright
The generation of T cells with maximal anti-tumor activities will significantly impact the field of T-cellbased adoptive immunotherapy. In this report, we found that OKT3/IL-2-stimulated T cells were phenotypically more heterogeneous, with enhanced anti-tumor activity in vivo and when locally administered in a solid tumor mouse model. To further improve the OKT3/IL-2-based T cell manufacturing procedure, we developed a novel T cell stimulation and expansion method in which peripheral blood mononuclear cells were electroporated with mRNA encoding a chimeric membrane protein consisting of a single-chain variable fragment against CD3 and the intracellular domains of CD28 and 4-1BB (OKT3-28BB). The expanded T cells were phenotypically and functionally similar to T cells expanded by OKT3/IL-2. Moreover, co-electroporation of CD86 and 4-1BBL could further change the phenotype and enhance the in vivo anti-tumor activity. Although T cells expanded by the coelectroporation of OKT3-28BB with CD86 and 4-1BBL showed an increased central memory phenotype, the T cells still maintained tumor lytic activities as potent as those of OKT3/IL-2 or OKT3-28BB-stimulated T cells. In different tumor mouse models, T cells expanded by OKT3-28BB RNA electroporation showed anti-tumor activities superior to those of OKT3/IL-2 T cells. Hence, T cells with both a less differentiated phenotype and potent tumor killing ability can be generated by RNA electroporation, and this T cell manufacturing procedure can be further optimized by simply co-delivering other splices of RNA, thus providing a simple and cost-effective method for generating high-quality T cells for adoptive immunotherapy.
Key words： T lymphocytes; CAR; manufacture; gene transfer; RNA electroporation
Xiaojun Liu , Shuguang Jiang , Chongyun Fang , Hua Li , Xuhua Zhang , Fuqin Zhang , Carl H. June , Yangbing Zhao . Novel T cells with improved in vivo anti-tumor activity generated by RNA electroporation[J]. Protein & Cell, 2017 , 8(7) : 514 -526 . DOI: 10.1007/s13238-017-0422-6
| 1 |
BarrettDM, ZhaoY, LiuX, JiangS, CarpenitoC, KalosM, CarrollRG, JuneCH, GruppSA (2011) Treatment of advanced leukemia in mice with mRNA engineered T cells.Human Gene Ther22:1575–1586
|
| 2 |
BarrettDM, SinghN, LiuX, JiangS, JuneCH, GruppSA, ZhaoY (2014) Relation of clinical culture method to T-cell memory status and efficacy in xenograft models of adoptive immunotherapy.Cytotherapy16:619–630
|
| 3 |
BesserMJ, Shapira-FrommerR, TrevesAJ, ZippelD, ItzhakiO, HershkovitzL, LevyD, KubiA, HovavE, ChermoshniukN
|
| 4 |
BrentjensRJ, DavilaML,RiviereI, ParkJ,WangX, CowellLG,BartidoS, StefanskiJ, TaylorC, OlszewskaM
|
| 5 |
ButlerMO, ImatakiO, YamashitaY, TanakaM, AnsenS, BerezovskayaA, MetzlerG, MilsteinMI, MooneyMM, MurrayAP
|
| 6 |
CampagnoloC, MeyersKJ, RyanT, AtkinsonRC, ChenYT, ScanlanMJ, RitterG, OldLJ, BattCA (2004) Real-Time, label-free monitoring of tumor antigen and serum antibody interactions.J Biochem Biophys Methods61:283–298
|
| 7 |
CarpenitoC, MiloneMC, HassanR, SimonetJC, LakhalM, SuhoskiMM, Varela-RohenaA, HainesKM, HeitjanDF, AlbeldaSM
|
| 8 |
GattinoniL, KlebanoffCA, PalmerDC, WrzesinskiC, KerstannK, YuZ, FinkelsteinSE, TheoretMR, RosenbergSA, RestifoNP (2005) Acquisition of full effector function in vitro paradoxically impairs the in vivo antitumor efficacy of adoptively transferred CD8+ T cells.J Clin Investig115:1616–1626
|
| 9 |
GattinoniL, LugliE, JiY, PosZ, PaulosCM, QuigleyMF, AlmeidaJR, GostickE, YuZ, CarpenitoC
|
| 10 |
GattinoniL, SpeiserDE, LichterfeldM, BoniniC (2017) T memory stem cells in health and disease.Nat Med23:18–27
|
| 11 |
HermansIF, SilkJD, YangJ, PalmowskiMJ, GileadiU, McCarthyC, SalioM, RoncheseF, CerundoloV (2004) The VITAL assay: a versatile fluorometric technique for assessing CTL- and NKTmediated cytotoxicity against multiple targets in vitro and in vivo.J Immunol Methods285:25–40
|
| 12 |
KlossCC, CondominesM, CartellieriM, BachmannM, SadelainM (2013) Combinatorial antigen recognition with balanced signaling promotes selective tumor eradication by engineered T cells.Nat Biotechnol31:71–75
|
| 13 |
LeeDW, KochenderferJN, Stetler-StevensonM, CuiYK, DelbrookC, FeldmanSA, FryTJ, OrentasR, SabatinoM, ShahNN
|
| 14 |
LevineBL, BernsteinWB, ConnorsM, CraigheadN, LindstenT, ThompsonCB, JuneCH (1997) Effects of CD28 costimulation on long-term proliferation of CD4+ T cells in the absence of exogenous feeder cells.J Immunol159:5921–5930
|
| 15 |
LiuX, JiangS, FangC, YangS, OlalereD, PequignotEC, CogdillAP, LiN, RamonesM, GrandaB
|
| 16 |
MausMV, FraiettaJA, LevineBL, KalosM, ZhaoY, JuneCH (2014) Adoptive immunotherapy for cancer or viruses.Ann Rev Immunol32:189–225
|
| 17 |
MorganRA, DudleyME, WunderlichJR, HughesMS, YangJC, SherryRM, RoyalRE, TopalianSL, KammulaUS, RestifoNP
|
| 18 |
PorterDL, LevineBL, KalosM, BaggA, JuneCH (2011) Chimeric antigen receptor-modified T cells in chronic lymphoid leukemia.N Engl J Med365:725–733
|
| 19 |
PowellDJ Jr, DudleyME, RobbinsPF, RosenbergSA (2005) Transition of late-stage effector T cells to CD27+ CD28+ tumorreactive effector memory T cells in humans after adoptive cell transfer therapy.Blood105:241–250
|
| 20 |
RenJ, LiuX, FangC, JiangS, JuneCH, ZhaoY (2016) Multiplex genome editing to generate universal CAR T cells resistant to PD1 inhibition.Clin Cancer Res23(9):2255–2266
|
| 21 |
RiddellSR, GreenbergPD (1990) The use of anti-CD3 and anti-CD28 monoclonal antibodies to clone and expand human antigen-specific T cells.J Immunol Methods128:189–201
|
| 22 |
RosenbergSA(2008) Overcoming obstacles to the effective immunotherapy of human cancer.Proc Natl Acad Sci USA105:12643–12644
|
| 23 |
SabatinoM, HuJ, SommarivaM, GautamS, FellowesV, HockerJD, DoughertyS, QinH, KlebanoffCA, FryTJ
|
| 24 |
StephanMT, PonomarevV, BrentjensRJ, ChangAH, DobrenkovKV, HellerG, SadelainM (2007) T cell-encoded CD80 and 4-1BBL induce auto- and transcostimulation, resulting in potent tumor rejection.Nat Med13:1440–1449
|
| 25 |
SuhoskiMM, GolovinaTN, AquiNA, TaiVC, Varela-RohenaA, MiloneMC, CarrollRG, RileyJL, JuneCH (2007) Engineering artificial antigen-presenting cells to express a diverse array of costimulatory molecules.Mol Ther15:981–988
|
| 26 |
WattsTH (2005)TNF/TNFR family members in costimulation of T cell responses.Ann Rev Immunol23:23–68
|
| 27 |
YangS, DudleyME, RosenbergSA, MorganRA (2010) A simplified method for the clinical-scale generation of central memory-like CD8+ T cells after transduction with lentiviral vectors encoding antitumor antigen T-cell receptors.J Immunother33:648–658
|
| 28 |
YangS, JiY, GattinoniL, ZhangL, YuZ, RestifoNP, RosenbergSA, MorganRA (2013) Modulating the differentiation status of ex vivo-cultured anti-tumor T cells using cytokine cocktails.Cancer Immunol Immunother62:727–736
|
| 29 |
ZhaoY, BoczkowskiD, NairSK, GilboaE (2003) Inhibition of invariant chain expression in dendritic cells presenting endogenous antigens stimulates CD4+ T-cell responses and tumor immunity.Blood102:4137–4142
|
| 30 |
ZhaoY, ZhengZ, CohenCJ, GattinoniL, PalmerDC, RestifoNP, RosenbergSA, MorganRA (2006) High-efficiency transfection of primary human and mouse T lymphocytes using RNA electroporation.Mol Ther13:151–159
|
| 31 |
ZhaoY, MoonE, CarpenitoC, PaulosCM, LiuX, BrennanAL, ChewA, CarrollRG, SchollerJ, LevineBL
|
| 32 |
ZhouJ, DudleyME, RosenbergSA, RobbinsPF (2005a) Persistence of multiple tumor-specific T-cell clones is associated with complete tumor regression in a melanoma patient receiving adoptive cell transfer therapy.J Immunother28:53–62
|
| 33 |
ZhouJ, ShenX, HuangJ, HodesRJ, RosenbergSA, RobbinsPF (2005b) Telomere length of transferred lymphocytes correlates with in vivo persistence and tumor regression in melanoma patients receiving cell transfer therapy.J Immunol175:7046–7052
|
/
| 〈 |
|
〉 |