RT Journal Article SR Electronic T1 Purinergic targeting enhances immunotherapy of CD73+ solid tumors with piggyBac-engineered chimeric antigen receptor natural killer cells JF Journal for ImmunoTherapy of Cancer JO J Immunother Cancer FD BMJ Publishing Group Ltd SP 136 DO 10.1186/s40425-018-0441-8 VO 6 IS 1 A1 Jiao Wang A1 Kyle B. Lupo A1 Andrea M. Chambers A1 Sandro Matosevic YR 2018 UL http://jitc.bmj.com/content/6/1/136.abstract AB Background The anti-tumor immunity of natural killer (NK) cells can be paralyzed by the CD73-induced generation of immunosuppressive adenosine from precursor ATP within the hypoxic microenvironment of solid tumors. In an effort to redirect purinergic immunosuppression of NK cell anti-tumor function, we showed, for the first time, that immunometabolic combination treatment with NKG2D-engineered CAR-NK cells alongside blockade of CD73 ectonucleotidase activity can result in significant anti-tumor responses in vivo.Methods NK cells were engineered non-virally with NKG2D.CAR-presenting vectors based on the piggyBac transposon system with DAP10 and CD3ζ co-signaling domains. The anti-tumor immunity of NKG2D.CAR.NK cells in combination with CD73 targeting was evaluated against multiple solid tumor targets in vitro and humanized mouse xenografts in immunodeficient tumor-bearing mice in vivo. Intratumoral migration was evaluated via immunohistochemical staining, while degranulation capacity and IFN-γ production of NK cells were measured in response to solid tumor targets.Results Our results showed that CD73 blockade can mediate effective purinergic reprogramming and enhance anti-tumor cytotoxicity both in vitro and in vivo by enhancing the killing ability of CAR-engineered NK cells against CD73+ solid tumor targets via mechanisms that might imply alleviation from adenosinergic immunometabolic suppression. CD73 blockade improved the intratumoral homing of CD56+ CAR-NK cells in vivo. These engineered NK cells showed synergistic therapeutic efficacy in combination with CD73 targeting against CD73+ human lung cancer xenograft models. Interestingly, CD73 blockade could inhibit tumor growth in vivo independently of adaptive immune cells, innate immunity or NK cell-mediated ADCC.Conclusions Immunotherapies targeting the adenosinergic signaling cascade, which act by neutralizing CD73 ectoenzymatic activity, had thus far not been evaluated in humanized tumor models, nor had the implication of innate immunity been investigated. Taken together, our pre-clinical efficacy data demonstrate, for the first time, the potential of targeting CD73 to modulate purinergic signaling and enhance adoptive NK cell immunotherapy via mechanisms that could implicate autocrine tumor control as well as by mediating adenosinergic signaling.Abbreviations:ADAAdenosine deaminaseADCCAntibody-dependent cellular cytotoxicityADOAdenosineADPAdenosine diphosphateAMPAdenosine monophosphateAMPrAmpicillin resistanceATPAdenosine triphosphateBGH PABovine growth hormone polyadenylation signalCAGChicken β actinCARChimeric antigen receptorCMVHuman cytomegalovirusECMExracellular matrixEF1AEukaryotic translation elongation factor 1 alpha 1EHNAErythro-9-(2-hydroxy-3-nonyl)adenineENTPDase1Ectonucleoside triphosphate diphosphohydrolase-1IFN-γInterferon-γMDSCMyeloid-derived suppressor cellMFIMean fluorescence intensityMHCMajor histocompatibility complexMMPMatrix metalloproteinaseNKNatural killerNKG2DNatural killer group 2 DNSGNon-obese diabetic scid gammaORIOrigin of replicationPBAEPoly(beta-amino ester)rBG pARabbit β-globin polyadenylation signalTMZTemozolomideTregRegulatory T cell