Granzyme A from cytotoxic lymphocytes cleaves GSDMB to trigger pyroptosis in target cells
Granzyme A lights a fire
Cytotoxic T cells and natural killer cells use several strategies to kill infected or transformed cells. One such pathway entails the delivery of a family of serine proteases called granzymes to target cells through perforin-mediated pores to induce a form of programmed cell death called apoptosis. Zhou et al. show that granzyme A cleaves and activates gasdermin B (GSDMB), a central player in the highly inflammatory cell death process known as pyroptosis (see the Perspective by Nicolai and Raulet). GSDMB expression was highly expressed in some tissues and could be up-regulated by interferon-γ. Enforced expression of GSDMB in cancer cells enhanced tumor clearance in a mouse model, suggesting that this pathway may be a target for future cancer immunotherapies.
Structured Abstract
INTRODUCTION
In cellular immunity, cytotoxic T lymphocytes (CTLs) and natural killer (NK) cells use perforin to deliver serine protease granzymes into target cells to kill them. Gasdermins are pore-forming proteins that execute pyroptosis, a form of proinflammatory cell death. Gasdermin D (GSDMD) is cleaved by caspase-1/4/5/11 upon inflammasome activation, releasing the pore-forming domain for plasma membrane disruption. Gasdermin E (GSDME) is similarly cleaved by caspase-3, converting apoptosis to pyroptosis. The functional mechanism for other gasdermins is unknown.
RATIONALE
The view that granzymes induce target-cell apoptosis was proposed two decades ago, when apoptosis was thought to be the dominant form of programmed cell death and assays to ascertain apoptosis were insufficiently accurate. Furthermore, granzyme cytotoxicity was only assessed in a few cell types. Discovery of the gasdermin family, which are true cell death executors, has altered our understanding of programmed cell death. In this work, we explored whether members of the gasdermin family might respond to granzymes and induce pyroptosis.
RESULTS
The expression of gasdermin B (GSDMB) but no other gasdermins in human embryonic kidney (HEK) 293T cells induced pyroptotic killing by NK cells, accompanied by an interdomain cleavage of GSDMB. These processes were blocked by inhibiting the perforin–granzyme pathway. In vitro profiling of all five human granzymes identified granzyme A (GZMA), which readily cleaved GSDMB, predominantly at Lys244 within the interdomain linker. This cleavage unmasked the pore-forming activity of GSDMB. GZMA, delivered into GSDMB-reconstituted cells by electroporation or perforin, induced extensive pyroptosis with interdomain cleavage of GSDMB. These effects were diminished by a K229A/K244A (KK/A) mutation of GSDMB [in which lysine (K) was replaced by alanine (A) at positions 229 and 224, respectively]. In cells normally undergoing apoptosis upon GZMA delivery, the additional expression of GZMA-cleavable GSDMB converted apoptosis into pyroptosis. Pyroptotic killing by NK cells was blocked by both the KK/AA mutation and a knockdown of GZMA expression. Among 39 cell lines, three, including the esophageal carcinoma (OE19 cells), expressed GSDMB and underwent pyroptosis upon GZMA delivery. Knockout experiments revealed that pyroptosis in OE19 cells required the interdomain cleavage of GSDMB. Furthermore, GSDMB expression was up-regulated by interferon-γ (IFN-γ). Approximately one-third of GSDMB-negative cell lines showed IFN-γ–induced GSDMB expression. IFN-γ promoted GZMA- or NK cell–induced pyroptosis in several target cells. Primary T cells, including anti-CD19 chimeric-antigen receptor (CAR) T cells and NY-ESO-1–specific T cell receptor (TCR)–engineered T cells (TCR T cells), also induced pyroptosis in GSDMB-reconstituted cells through cleavage of GSDMB by GZMA. Introducing GZMA-cleavable GSDMB into mouse cancer cells promoted tumor clearance in mice. GSDMB was highly expressed in certain tissues, particularly digestive tract epithelia, including the derived tumors. GSDMB appeared to be silenced in gastric and esophageal cancers. The Cancer Genome Atlas database recorded a strong positive correlation between GSDMB expression and patient survival for bladder carcinoma and skin cutaneous melanoma.
CONCLUSION
GZMA from cytotoxic lymphocytes cleaves and activates GSDMB to induce target cell pyroptosis. This immune effector mechanism promotes CTL-mediated tumor clearance in mice. High GSDMB expression in the digestive system suggests the importance of GSDMB-mediated immunity in these tissues and will guide immunotherapy for related cancers. Our findings suggest that substrates such as gasdermins, rather than their upstream proteases, determine the nature of cell death.
GZMA from cytotoxic lymphocytes cleaves GSDMB in target cells, predominantly at Lys244 within the interdomain linker.
The cleavage allows GSDMB pore-forming domain (GSDMB-N) to perforate plasma membrane and induce pyroptosis. Expression of GSDMB wild type (WT) but not its GZMA-resistant K/A mutant in mouse cancer cells promotes cytotoxic T lymphocyte–mediated tumor clearance when the inhibitory checkpoint is blocked by antibody to programmed cell death 1 (PD-1). IFNGR, IFN-γ receptor.
Abstract
Cytotoxic lymphocyte–mediated immunity relies on granzymes. Granzymes are thought to kill target cells by inducing apoptosis, although the underlying mechanisms are not fully understood. Here, we report that natural killer cells and cytotoxic T lymphocytes kill gasdermin B (GSDMB)–positive cells through pyroptosis, a form of proinflammatory cell death executed by the gasdermin family of pore-forming proteins. Killing results from the cleavage of GSDMB by lymphocyte-derived granzyme A (GZMA), which unleashes its pore-forming activity. Interferon-γ (IFN-γ) up-regulates GSDMB expression and promotes pyroptosis. GSDMB is highly expressed in certain tissues, particularly digestive tract epithelia, including derived tumors. Introducing GZMA-cleavable GSDMB into mouse cancer cells promotes tumor clearance in mice. This study establishes gasdermin-mediated pyroptosis as a cytotoxic lymphocyte–killing mechanism, which may enhance antitumor immunity.
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Supplementary Material
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References and Notes
1
C. L. Ewen, K. P. Kane, R. C. Bleackley, A quarter century of granzymes. Cell Death Differ. 19, 28–35 (2012).
2
I. Voskoboinik, J. C. Whisstock, J. A. Trapani, Perforin and granzymes: Function, dysfunction and human pathology. Nat. Rev. Immunol. 15, 388–400 (2015).
3
J. W. Heusel, R. L. Wesselschmidt, S. Shresta, J. H. Russell, T. J. Ley, Cytotoxic lymphocytes require granzyme B for the rapid induction of DNA fragmentation and apoptosis in allogeneic target cells. Cell 76, 977–987 (1994).
4
D. Kägi, F. Vignaux, B. Ledermann, K. Bürki, V. Depraetere, S. Nagata, H. Hengartner, P. Golstein, Fas and perforin pathways as major mechanisms of T cell-mediated cytotoxicity. Science 265, 528–530 (1994).
5
B. Lowin, M. Hahne, C. Mattmann, J. Tschopp, Cytolytic T-cell cytotoxicity is mediated through perforin and Fas lytic pathways. Nature 370, 650–652 (1994).
6
A. J. Darmon, D. W. Nicholson, R. C. Bleackley, Activation of the apoptotic protease CPP32 by cytotoxic T-cell-derived granzyme B. Nature 377, 446–448 (1995).
7
J. H. Russell, T. J. Ley, Lymphocyte-mediated cytotoxicity. Annu. Rev. Immunol. 20, 323–370 (2002).
8
J. A. Trapani, M. J. Smyth, Functional significance of the perforin/granzyme cell death pathway. Nat. Rev. Immunol. 2, 735–747 (2002).
9
Y. Oshimi, K. Oshimi, S. Miyazaki, Necrosis and apoptosis associated with distinct Ca2+ response patterns in target cells attacked by human natural killer cells. J. Physiol. 495, 319–329 (1996).
10
S. S. Somanchi, K. J. McCulley, A. Somanchi, L. L. Chan, D. A. Lee, A novel method for assessment of natural killer cell cytotoxicity using image cytometry. PLOS ONE 10, e0141074 (2015).
11
C. S. Backes, K. S. Friedmann, S. Mang, A. Knörck, M. Hoth, C. Kummerow, Natural killer cells induce distinct modes of cancer cell death: Discrimination, quantification, and modulation of apoptosis, necrosis, and mixed forms. J. Biol. Chem. 293, 16348–16363 (2018).
12
A. Zychlinsky, L. M. Zheng, C. C. Liu, J. D. Young, Cytolytic lymphocytes induce both apoptosis and necrosis in target cells. J. Immunol. 146, 393–400 (1991).
13
D. K. Blanchard, S. Wei, C. Duan, F. Pericle, J. I. Diaz, J. Y. Djeu, Role of extracellular adenosine triphosphate in the cytotoxic T-lymphocyte-mediated lysis of antigen presenting cells. Blood 85, 3173–3182 (1995).
14
N. J. Waterhouse, V. R. Sutton, K. A. Sedelies, A. Ciccone, M. Jenkins, S. J. Turner, P. I. Bird, J. A. Trapani, Cytotoxic T lymphocyte-induced killing in the absence of granzymes A and B is unique and distinct from both apoptosis and perforin-dependent lysis. J. Cell Biol. 173, 133–144 (2006).
15
P. A. Henkart, M. S. Williams, C. M. Zacharchuk, A. Sarin, Do CTL kill target cells by inducing apoptosis? Semin. Immunol. 9, 135–144 (1997).
16
D. Chowdhury, J. Lieberman, Death by a thousand cuts: Granzyme pathways of programmed cell death. Annu. Rev. Immunol. 26, 389–420 (2008).
17
J. Lieberman, Granzyme A activates another way to die. Immunol. Rev. 235, 93–104 (2010).
18
K. Ebnet, M. Hausmann, F. Lehmann-Grube, A. Müllbacher, M. Kopf, M. Lamers, M. M. Simon, Granzyme A-deficient mice retain potent cell-mediated cytotoxicity. EMBO J. 14, 4230–4239 (1995).
19
O. Susanto, S. E. Stewart, I. Voskoboinik, D. Brasacchio, M. Hagn, S. Ellis, S. Asquith, K. A. Sedelies, P. I. Bird, N. J. Waterhouse, J. A. Trapani, Mouse granzyme A induces a novel death with writhing morphology that is mechanistically distinct from granzyme B-induced apoptosis. Cell Death Differ. 20, 1183–1193 (2013).
20
T. Bergsbaken, S. L. Fink, B. T. Cookson, Pyroptosis: Host cell death and inflammation. Nat. Rev. Microbiol. 7, 99–109 (2009).
21
N. Kayagaki, S. Warming, M. Lamkanfi, L. Vande Walle, S. Louie, J. Dong, K. Newton, Y. Qu, J. Liu, S. Heldens, J. Zhang, W. P. Lee, M. Roose-Girma, V. M. Dixit, Non-canonical inflammasome activation targets caspase-11. Nature 479, 117–121 (2011).
22
J. Shi, Y. Zhao, Y. Wang, W. Gao, J. Ding, P. Li, L. Hu, F. Shao, Inflammatory caspases are innate immune receptors for intracellular LPS. Nature 514, 187–192 (2014).
23
N. Kayagaki, I. B. Stowe, B. L. Lee, K. O’Rourke, K. Anderson, S. Warming, T. Cuellar, B. Haley, M. Roose-Girma, Q. T. Phung, P. S. Liu, J. R. Lill, H. Li, J. Wu, S. Kummerfeld, J. Zhang, W. P. Lee, S. J. Snipas, G. S. Salvesen, L. X. Morris, L. Fitzgerald, Y. Zhang, E. M. Bertram, C. C. Goodnow, V. M. Dixit, Caspase-11 cleaves gasdermin D for non-canonical inflammasome signalling. Nature 526, 666–671 (2015).
24
J. Shi, Y. Zhao, K. Wang, X. Shi, Y. Wang, H. Huang, Y. Zhuang, T. Cai, F. Wang, F. Shao, Cleavage of GSDMD by inflammatory caspases determines pyroptotic cell death. Nature 526, 660–665 (2015).
25
J. Shi, W. Gao, F. Shao, Pyroptosis: Gasdermin-Mediated Programmed Necrotic Cell Death. Trends Biochem. Sci. 42, 245–254 (2017).
26
L. Galluzzi, I. Vitale, S. A. Aaronson, J. M. Abrams, D. Adam, P. Agostinis, E. S. Alnemri, L. Altucci, I. Amelio, D. W. Andrews, M. Annicchiarico-Petruzzelli, A. V. Antonov, E. Arama, E. H. Baehrecke, N. A. Barlev, N. G. Bazan, F. Bernassola, M. J. M. Bertrand, K. Bianchi, M. V. Blagosklonny, K. Blomgren, C. Borner, P. Boya, C. Brenner, M. Campanella, E. Candi, D. Carmona-Gutierrez, F. Cecconi, F. K.-M. Chan, N. S. Chandel, E. H. Cheng, J. E. Chipuk, J. A. Cidlowski, A. Ciechanover, G. M. Cohen, M. Conrad, J. R. Cubillos-Ruiz, P. E. Czabotar, V. D’Angiolella, T. M. Dawson, V. L. Dawson, V. De Laurenzi, R. De Maria, K.-M. Debatin, R. J. DeBerardinis, M. Deshmukh, N. Di Daniele, F. Di Virgilio, V. M. Dixit, S. J. Dixon, C. S. Duckett, B. D. Dynlacht, W. S. El-Deiry, J. W. Elrod, G. M. Fimia, S. Fulda, A. J. García-Sáez, A. D. Garg, C. Garrido, E. Gavathiotis, P. Golstein, E. Gottlieb, D. R. Green, L. A. Greene, H. Gronemeyer, A. Gross, G. Hajnoczky, J. M. Hardwick, I. S. Harris, M. O. Hengartner, C. Hetz, H. Ichijo, M. Jäättelä, B. Joseph, P. J. Jost, P. P. Juin, W. J. Kaiser, M. Karin, T. Kaufmann, O. Kepp, A. Kimchi, R. N. Kitsis, D. J. Klionsky, R. A. Knight, S. Kumar, S. W. Lee, J. J. Lemasters, B. Levine, A. Linkermann, S. A. Lipton, R. A. Lockshin, C. López-Otín, S. W. Lowe, T. Luedde, E. Lugli, M. MacFarlane, F. Madeo, M. Malewicz, W. Malorni, G. Manic, J.-C. Marine, S. J. Martin, J.-C. Martinou, J. P. Medema, P. Mehlen, P. Meier, S. Melino, E. A. Miao, J. D. Molkentin, U. M. Moll, C. Muñoz-Pinedo, S. Nagata, G. Nuñez, A. Oberst, M. Oren, M. Overholtzer, M. Pagano, T. Panaretakis, M. Pasparakis, J. M. Penninger, D. M. Pereira, S. Pervaiz, M. E. Peter, M. Piacentini, P. Pinton, J. H. M. Prehn, H. Puthalakath, G. A. Rabinovich, M. Rehm, R. Rizzuto, C. M. P. Rodrigues, D. C. Rubinsztein, T. Rudel, K. M. Ryan, E. Sayan, L. Scorrano, F. Shao, Y. Shi, J. Silke, H.-U. Simon, A. Sistigu, B. R. Stockwell, A. Strasser, G. Szabadkai, S. W. G. Tait, D. Tang, N. Tavernarakis, A. Thorburn, Y. Tsujimoto, B. Turk, T. Vanden Berghe, P. Vandenabeele, M. G. Vander Heiden, A. Villunger, H. W. Virgin, K. H. Vousden, D. Vucic, E. F. Wagner, H. Walczak, D. Wallach, Y. Wang, J. A. Wells, W. Wood, J. Yuan, Z. Zakeri, B. Zhivotovsky, L. Zitvogel, G. Melino, G. Kroemer, Molecular mechanisms of cell death: Recommendations of the Nomenclature Committee on Cell Death 2018. Cell Death Differ. 25, 486–541 (2018).
27
R. A. Aglietti, A. Estevez, A. Gupta, M. G. Ramirez, P. S. Liu, N. Kayagaki, C. Ciferri, V. M. Dixit, E. C. Dueber, GsdmD p30 elicited by caspase-11 during pyroptosis forms pores in membranes. Proc. Natl. Acad. Sci. U.S.A. 113, 7858–7863 (2016).
28
J. Ding, K. Wang, W. Liu, Y. She, Q. Sun, J. Shi, H. Sun, D.-C. Wang, F. Shao, Pore-forming activity and structural autoinhibition of the gasdermin family. Nature 535, 111–116 (2016).
29
X. Liu, Z. Zhang, J. Ruan, Y. Pan, V. G. Magupalli, H. Wu, J. Lieberman, Inflammasome-activated gasdermin D causes pyroptosis by forming membrane pores. Nature 535, 153–158 (2016).
30
L. Sborgi, S. Rühl, E. Mulvihill, J. Pipercevic, R. Heilig, H. Stahlberg, C. J. Farady, D. J. Müller, P. Broz, S. Hiller, GSDMD membrane pore formation constitutes the mechanism of pyroptotic cell death. EMBO J. 35, 1766–1778 (2016).
31
P. Broz, P. Pelegrín, F. Shao, The gasdermins, a protein family executing cell death and inflammation. Nat. Rev. Immunol. 20, 143–157 (2020).
32
C. Rogers, T. Fernandes-Alnemri, L. Mayes, D. Alnemri, G. Cingolani, E. S. Alnemri, Cleavage of DFNA5 by caspase-3 during apoptosis mediates progression to secondary necrotic/pyroptotic cell death. Nat. Commun. 8, 14128 (2017).
33
Y. Wang, W. Gao, X. Shi, J. Ding, W. Liu, H. He, K. Wang, F. Shao, Chemotherapy drugs induce pyroptosis through caspase-3 cleavage of a gasdermin. Nature 547, 99–103 (2017).
34
K. Akino, M. Toyota, H. Suzuki, T. Imai, R. Maruyama, M. Kusano, N. Nishikawa, Y. Watanabe, Y. Sasaki, T. Abe, E. Yamamoto, I. Tarasawa, T. Sonoda, M. Mori, K. Imai, Y. Shinomura, T. Tokino, Identification of DFNA5 as a target of epigenetic inactivation in gastric cancer. Cancer Sci. 98, 88–95 (2007).
35
M. S. Kim, X. Chang, K. Yamashita, J. K. Nagpal, J. H. Baek, G. Wu, B. Trink, E. A. Ratovitski, M. Mori, D. Sidransky, Aberrant promoter methylation and tumor suppressive activity of the DFNA5 gene in colorectal carcinoma. Oncogene 27, 3624–3634 (2008).
36
K. Yokomizo, Y. Harada, K. Kijima, K. Shinmura, M. Sakata, K. Sakuraba, Y. Kitamura, A. Shirahata, T. Goto, H. Mizukami, M. Saito, G. Kigawa, H. Nemoto, K. Hibi, Methylation of the DFNA5 gene is frequently detected in colorectal cancer. Anticancer Res. 32, 1319–1322 (2012).
37
C. J. Wang, L. Tang, D.-W. Shen, C. Wang, Q.-Y. Yuan, W. Gao, Y.-K. Wang, R.-H. Xu, H. Zhang, The expression and regulation of DFNA5 in human hepatocellular carcinoma DFNA5 in hepatocellular carcinoma. Mol. Biol. Rep. 40, 6525–6531 (2013).
38
B. Vanherberghen, P. E. Olofsson, E. Forslund, M. Sternberg-Simon, M. A. Khorshidi, S. Pacouret, K. Guldevall, M. Enqvist, K.-J. Malmberg, R. Mehr, B. Önfelt, Classification of human natural killer cells based on migration behavior and cytotoxic response. Blood 121, 1326–1334 (2013).
39
R. Roger, J. Bréard, M. Comisso, C. Bohuon, M. Pallardy, J. Bertoglio, CD28-mediated cytotoxicity of YT natural killer cells on B7-positive targets induces rapid necrotic death independent of granule exocytosis. Cell. Immunol. 168, 24–32 (1996).
40
S. Mahrus, C. S. Craik, Selective chemical functional probes of granzymes A and B reveal granzyme B is a major effector of natural killer cell-mediated lysis of target cells. Chem. Biol. 12, 567–577 (2005).
41
T. Zhao, H. Zhang, Y. Guo, Q. Zhang, G. Hua, H. Lu, Q. Hou, H. Liu, Z. Fan, Granzyme K cleaves the nucleosome assembly protein SET to induce single-stranded DNA nicks of target cells. Cell Death Differ. 14, 489–499 (2007).
42
P. J. Beresford, C. M. Kam, J. C. Powers, J. Lieberman, Recombinant human granzyme A binds to two putative HLA-associated proteins and cleaves one of them. Proc. Natl. Acad. Sci. U.S.A. 94, 9285–9290 (1997).
43
D. A. Thomas, C. Du, M. Xu, X. Wang, T. J. Ley, DFF45/ICAD can be directly processed by granzyme B during the induction of apoptosis. Immunity 12, 621–632 (2000).
44
C. L. Ewen, K. P. Kane, R. C. Bleackley, Granzyme H induces cell death primarily via a Bcl-2-sensitive mitochondrial cell death pathway that does not require direct Bid activation. Mol. Immunol. 54, 309–318 (2013).
45
H. Lu, Q. Hou, T. Zhao, H. Zhang, Q. Zhang, L. Wu, Z. Fan, Granzyme M directly cleaves inhibitor of caspase-activated DNase (CAD) to unleash CAD leading to DNA fragmentation. J. Immunol. 177, 1171–1178 (2006).
46
K. Sakuishi, L. Apetoh, J. M. Sullivan, B. R. Blazar, V. K. Kuchroo, A. C. Anderson, Targeting Tim-3 and PD-1 pathways to reverse T cell exhaustion and restore anti-tumor immunity. J. Exp. Med. 207, 2187–2194 (2010).
47
S. I. Mosely, J. E. Prime, R. C. A. Sainson, J.-O. Koopmann, D. Y. Q. Wang, D. M. Greenawalt, M. J. Ahdesmaki, R. Leyland, S. Mullins, L. Pacelli, D. Marcus, J. Anderton, A. Watkins, J. Coates Ulrichsen, P. Brohawn, B. W. Higgs, M. McCourt, H. Jones, J. A. Harper, M. Morrow, V. Valge-Archer, R. Stewart, S. J. Dovedi, R. W. Wilkinson, Rational selection of syngeneic preclinical tumor models for immunotherapeutic drug discovery. Cancer Immunol. Res. 5, 29–41 (2017).
48
A. O. Kamphorst, A. Wieland, T. Nasti, S. Yang, R. Zhang, D. L. Barber, B. T. Konieczny, C. Z. Daugherty, L. Koenig, K. Yu, G. L. Sica, A. H. Sharpe, G. J. Freeman, B. R. Blazar, L. A. Turka, T. K. Owonikoko, R. N. Pillai, S. S. Ramalingam, K. Araki, R. Ahmed, Rescue of exhausted CD8 T cells by PD-1-targeted therapies is CD28-dependent. Science 355, 1423–1427 (2017).
49
F. Arce Vargas, A. J. S. Furness, I. Solomon, K. Joshi, L. Mekkaoui, M. H. Lesko, E. Miranda Rota, R. Dahan, A. Georgiou, A. Sledzinska, A. Ben Aissa, D. Franz, M. Werner Sunderland, Y. N. S. Wong, J. Y. Henry, T. O’Brien, D. Nicol, B. Challacombe, S. A. Beers, S. Turajlic, M. Gore, J. Larkin, C. Swanton, K. A. Chester, M. Pule, J. V. Ravetch, T. Marafioti, K. S. Peggs, S. A. Quezada, Melanoma TRACERx Consortium; Renal TRACERx Consortium, Lung TRACERx Consortium, Fc-optimized anti-CD25 depletes tumor-infiltrating regulatory T cells and synergizes with PD-1 blockade to eradicate established tumors. Immunity 46, 577–586 (2017).
50
N. Saeki, T. Usui, K. Aoyagi, D. H. Kim, M. Sato, T. Mabuchi, K. Yanagihara, K. Ogawa, H. Sakamoto, T. Yoshida, H. Sasaki, Distinctive expression and function of four GSDM family genes (GSDMA-D) in normal and malignant upper gastrointestinal epithelium. Genes Chromosomes Cancer 48, 261–271 (2009).
51
Z. Tang, B. Kang, C. Li, T. Chen, Z. Zhang, GEPIA2: An enhanced web server for large-scale expression profiling and interactive analysis. Nucleic Acids Res. 47 (W1), W556–W560 (2019).
52
C. J. Froelich, J. Pardo, M. M. Simon, Granule-associated serine proteases: Granzymes might not just be killer proteases. Trends Immunol. 30, 117–123 (2009).
53
S. S. Metkar, C. Menaa, J. Pardo, B. Wang, R. Wallich, M. Freudenberg, S. Kim, S. M. Raja, L. Shi, M. M. Simon, C. J. Froelich, Human and mouse granzyme A induce a proinflammatory cytokine response. Immunity 29, 720–733 (2008).
54
P. F. Robbins, Y. F. Li, M. El-Gamil, Y. Zhao, J. A. Wargo, Z. Zheng, H. Xu, R. A. Morgan, S. A. Feldman, L. A. Johnson, A. D. Bennett, S. M. Dunn, T. M. Mahon, B. K. Jakobsen, S. A. Rosenberg, Single and dual amino acid substitutions in TCR CDRs can enhance antigen-specific T cell functions. J. Immunol. 180, 6116–6131 (2008).
55
C. M. Wang, Z.-Q. Wu, Y. Wang, Y.-L. Guo, H.-R. Dai, X.-H. Wang, X. Li, Y.-J. Zhang, W.-Y. Zhang, M.-X. Chen, Y. Zhang, K.-C. Feng, Y. Liu, S.-X. Li, Q.-M. Yang, W.-D. Han, Autologous T cells expressing CD30 chimeric antigen receptors for relapsed or refractory hodgkin lymphoma: An open-label phase I trial. Clin. Cancer Res. 23, 1156–1166 (2017).
56
K. C. Feng, Y. L. Guo, Y. Liu, H. R. Dai, Y. Wang, H. Y. Lv, J. H. Huang, Q. M. Yang, W. D. Han, Cocktail treatment with EGFR-specific and CD133-specific chimeric antigen receptor-modified T cells in a patient with advanced cholangiocarcinoma. J. Hematol. Oncol. 10, 4 (2017).
57
Y. Wang, W. Y. Zhang, Q. W. Han, Y. Liu, H. R. Dai, Y. L. Guo, J. Bo, H. Fan, Y. Zhang, Y. J. Zhang, M. X. Chen, K. C. Feng, Q. S. Wang, X. B. Fu, W. D. Han, Effective response and delayed toxicities of refractory advanced diffuse large B-cell lymphoma treated by CD20-directed chimeric antigen receptor-modified T cells. Clin. Immunol. 155, 160–175 (2014).
58
C. Li, W. Li, J. Xiao, S. Jiao, F. Teng, S. Xue, C. Zhang, C. Sheng, Q. Leng, C. E. Rudd, B. Wei, H. Wang, ADAP and SKAP55 deficiency suppresses PD-1 expression in CD8+ cytotoxic T lymphocytes for enhanced anti-tumor immunotherapy. EMBO Mol. Med. 7, 754–769 (2015).
59
F. Dotiwala, I. Fellay, L. Filgueira, D. Martinvalet, J. Lieberman, M. Walch, A high yield and cost-efficient expression system of human granzymes in mammalian cells. J. Vis. Exp. 10, e52911 (2015).
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Volume 368 | Issue 6494
29 May 2020
29 May 2020
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Received: 7 October 2019
Accepted: 3 April 2020
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Acknowledgments
We thank Z. Shen for providing antibody to PD-1 and NK-92MI and A375 cells, Z. Dong for YTS cells, X. Zhao for SW837 and SKCO1 cells, W. Shen for Edman sequencing, Y. Sun for assistance with microscopy, and Y. Xu for assistances with mouse experiments. We also thank members of the Shao laboratory for their assistance and helpful discussions. Funding: The work was supported by the Basic Science Center Project of the National Natural Science Foundation of China (81788104), the National Key Research and Development Program of China (2017YFA0505900 and 2016YFA0501500), the Chinese Academy of Medical Sciences Innovation Fund for Medical Sciences (2019-I2M-5-084), the Strategic Priority Research Program of the Chinese Academy of Sciences (XDB37030202), and a fund from Hoffmann-La Roche AG (ROADS027). Author contributions: Z.Z. and F.S. conceived the study; Z.Z., together with H.H. and Y.S., performed most of the experiments; K.W. contributed to the finding of GSDMB transcriptional priming; X.S., Yu.W., D.L., W.L., and J.D provided technical supports and insights. Ya.W. and W.H. prepared CAR T cells; Y.Z. and Lia.S. provided TCR T cells; Lin.S. provided tumor tissue samples; and Z.Z and F.S. analyzed the data and wrote the manuscript. F.S. obtained the funding and supervised the study. Competing interests: The authors declare no competing interests. Data and materials availability: All data are available in the main text or the supplementary materials. All the plasmids and cell lines generated in this study are available from the authors under a materials transfer agreement with the National Institute for Biological Sciences.
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Chinese Academy of Sciences: XDB08020202
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