The potential utility of immunotoxins for cancer therapy has convincingly been demonstrated in clinical studies. Nevertheless, the high immunogenicity of their bacterial toxin domain represents a critical limitation, and has prompted the evaluation of cell-death inducing proteins of human origin as a basis for less immunogenic immunotoxin-like molecules. In this review, we focus on the current status and future prospects of targeted fusion proteins for cancer therapy that employ granzyme B (GrB) from cytotoxic lymphocytes as a cytotoxic moiety. Naturally, this serine protease plays a critical role in the immune defense by inducing apoptotic target cell death upon cleavage of intracellular substrates. Advances in understanding of the structure and function of GrB enabled the generation of chimeric fusion proteins that carry a heterologous cell binding domain for recognition of tumor-associated cell surface antigens. These hybrid molecules display high selectivity for cancer cells, with cell killing activities similar to that of corresponding recombinant toxins. Recent findings have helped to understand and circumvent intrinsic cell binding of GrB and susceptibility of the enzyme to inhibition by serpins. This now allows the rational design of optimized GrB derivatives that avoid sequestration by binding to non-target tissues, limit off-target effects, and overcome resistance mechanisms in tumor cells.
References
[1]
Maloney, D.G. Immunotherapy for non-Hodgkin's lymphoma: Monoclonal antibodies and vaccines. J. Clin. Oncol. 2005, 23, 6421–6428, doi:10.1200/JCO.2005.06.004.
Hynes, N.E.; Lane, H.A. ERBB receptors and cancer: The complexity of targeted inhibitors. Nat. Rev. Cancer 2005, 5, 341–354, doi:10.1038/nrc1609.
[4]
Lan, K.H.; Lu, C.H.; Yu, D. Mechanisms of trastuzumab resistance and their clinical implications. Ann. NY Acad. Sci. 2005, 1059, 70–75, doi:10.1196/annals.1339.026.
[5]
Nahta, R.; Esteva, F.J. HER2 therapy: molecular mechanisms of trastuzumab resistance. Breast Cancer Res. 2006, 8, 215, doi:10.1186/bcr1612.
[6]
Weldon, J.E.; Pastan, I. A guide to taming a toxin--recombinant immunotoxins constructed from Pseudomonas exotoxin A for the treatment of cancer. FEBS J. 2011, 278, 4683–4700, doi:10.1111/j.1742-4658.2011.08182.x.
[7]
Wels, W.; Biburger, M.; Müller, T.; D？lken, B.; Giesübel, U.; Tonn, T.; Uherek, C. Recombinant immunotoxins and retargeted killer cells: employing engineered antibody fragments for tumor-specific targeting of cytotoxic effectors. Cancer Immunol. Immunother. 2004, 53, 217–226, doi:10.1007/s00262-003-0482-8.
[8]
Pastan, I.; Hassan, R.; Fitzgerald, D.J.; Kreitman, R.J. Immunotoxin therapy of cancer. Nat. Rev. Cancer 2006, 6, 559–565, doi:10.1038/nrc1891.
[9]
von Minckwitz, G.; Harder, S.; Hovelmann, S.; J？ger, E.; Al-Batran, S.E.; Loibl, S.; Atmaca, A.; Cimpoiasu, C.; Neumann, A.; Abera, A.; et al. Phase I clinical study of the recombinant antibody toxin scFv(FRP5)-ETA specific for the ErbB2/HER2 receptor in patients with advanced solid malignomas. Breast Cancer Res. 2005, 7, R617–R626.
[10]
Kreitman, R.J.; Pastan, I. Antibody fusion proteins: Anti-CD22 recombinant immunotoxin moxetumomab pasudotox. Clin. Cancer Res. 2011, 17, 6398–6405, doi:10.1158/1078-0432.CCR-11-0487.
[11]
Kelly, R.J.; Sharon, E.; Pastan, I.; Hassan, R. Mesothelin-targeted agents in clinical trials and in preclinical development. Mol. Cancer Ther. 2012, 11, 517–525, doi:10.1158/1535-7163.MCT-11-0454.
[12]
Pai, L.H.; Wittes, R.; Setser, A.; Willingham, M.C.; Pastan, I. Treatment of advanced solid tumors with immunotoxin LMB-1: An antibody linked to Pseudomonas exotoxin. Nat. Med. 1996, 2, 350–353, doi:10.1038/nm0396-350.
[13]
Azemar, M.; Djahansouzi, S.; J？ger, E.; Solbach, C.; Schmidt, M.; Maurer, A.B.; Mross, K.; Unger, C.; von Minckwitz, G.; Dall, P.; et al. Regression of cutaneous tumor lesions in patients intratumorally injected with a recombinant single-chain antibody-toxin targeted to ErbB2/HER2. Breast Cancer Res. Treat. 2003, 82, 155–164.
[14]
Keppler-Hafkemeyer, A.; Kreitman, R.J.; Pastan, I. Apoptosis induced by immunotoxins used in the treatment of hematologic malignancies. Int. J. Cancer 2000, 87, 86–94, doi:10.1002/1097-0215(20000701)87:1<86::AID-IJC13>3.0.CO;2-I.
[15]
Schmidt, M.; McWatters, A.; White, R.A.; Groner, B.; Wels, W.; Fan, Z.; Bast, R.C., Jr. Synergistic interaction between an anti-p185HER-2 pseudomonas exotoxin fusion protein [scFv(FRP5)-ETA] and ionizing radiation for inhibiting growth of ovarian cancer cells that overexpress HER-2. Gynecol. Oncol. 2001, 80, 145–155, doi:10.1006/gyno.2000.6040.
[16]
Weidle, U.H.; Georges, G.; Brinkmann, U. Fully human targeted cytotoxic fusion proteins: New anticancer agents on the horizon. Cancer Genomics Proteomics 2012, 9, 119–133.
[17]
Gerspach, J.; Wajant, H.; Pfizenmaier, K. Death ligands designed to kill: development and application of targeted cancer therapeutics based on proapoptotic TNF family ligands. Results Probl. Cell Differ. 2009, 49, 241–273, doi:10.1007/400_2008_22.
[18]
Antignani, A.; Youle, R.J. A chimeric protein induces tumor cell apoptosis by delivering the human Bcl-2 family BH3-only protein Bad. Biochemistry 2005, 44, 4074–4082, doi:10.1021/bi0477687.
[19]
Lorberboum-Galski, H. Human toxin-based recombinant immunotoxins/chimeric proteins as a drug delivery system for targeted treatment of human diseases. Expert Opin. Drug Deliv. 2011, 8, 605–621, doi:10.1517/17425247.2011.566269.
[20]
Hanahan, D.; Weinberg, R.A. Hallmarks of cancer: the next generation. Cell 2011, 144, 646–674, doi:10.1016/j.cell.2011.02.013.
[21]
Mahmud, H.; D？lken, B.; Wels, W.S. Induction of programmed cell death in ErbB2/HER2-expressing cancer cells by targeted delivery of apoptosis-inducing factor. Mol. Cancer Ther. 2009, 8, 1526–1535, doi:10.1158/1535-7163.MCT-08-1149.
[22]
Liu, Y.; Cheung, L.H.; Thorpe, P.; Rosenblum, M.G. Mechanistic studies of a novel human fusion toxin composed of vascular endothelial growth factor (VEGF)121 and the serine protease granzyme B: Directed apoptotic events in vascular endothelial cells. Mol. Cancer Ther. 2003, 2, 949–959.
[23]
D？lken, B.; Giesübel, U.; Knauer, S.K.; Wels, W.S. Targeted induction of apoptosis by chimeric granzyme B fusion proteins carrying antibody and growth factor domains for cell recognition. Cell Death Differ. 2006, 13, 576–585, doi:10.1038/sj.cdd.4401773.
[24]
Rosenblum, M.G.; Barth, S. Development of novel, highly cytotoxic fusion constructs containing granzyme B: unique mechanisms and functions. Curr. Pharm. Des. 2009, 15, 2676–2692, doi:10.2174/138161209788923958.
[25]
Kurschus, F.C.; Jenne, D.E. Delivery and therapeutic potential of human granzyme B. Immunol. Rev. 2010, 235, 159–171.
[26]
Cullen, S.P.; Brunet, M.; Martin, S.J. Granzymes in cancer and immunity. Cell Death Differ. 2010, 17, 616–623, doi:10.1038/cdd.2009.206.
[27]
Pham, C.T.; Ley, T.J. Dipeptidyl peptidase I is required for the processing and activation of granzymes A and B in vivo. Proc. Natl. Acad. Sci. USA 1999, 96, 8627–8632, doi:10.1073/pnas.96.15.8627.
[28]
Chowdhury, D.; Lieberman, J. Death by a thousand cuts: Granzyme pathways of programmed cell death. Annu. Rev. Immunol. 2008, 26, 389–420, doi:10.1146/annurev.immunol.26.021607.090404.
Huse, M.; Quann, E.J.; Davis, M.M. Shouts, whispers and the kiss of death: Directional secretion in T cells. Nat. Immunol. 2008, 9, 1105–1111, doi:10.1038/ni.f.215.
[31]
Afonina, I.S.; Cullen, S.P.; Martin, S.J. Cytotoxic and non-cytotoxic roles of the CTL/NK protease granzyme B. Immunol. Rev. 2010, 235, 105–116.
[32]
Baran, K.; Dunstone, M.; Chia, J.; Ciccone, A.; Browne, K.A.; Clarke, C.J.; Lukoyanova, N.; Saibil, H.; Whisstock, J.C.; Voskoboinik, I.; et al. The molecular basis for perforin oligomerization and transmembrane pore assembly. Immunity 2009, 30, 684–695, doi:10.1016/j.immuni.2009.03.016.
[33]
Law, R.H.; Lukoyanova, N.; Voskoboinik, I.; Caradoc-Davies, T.T.; Baran, K.; Dunstone, M.A.; D'Angelo, M.E.; Orlova, E.V.; Coulibaly, F.; Verschoor, S.; et al. The structural basis for membrane binding and pore formation by lymphocyte perforin. Nature 2010, 468, 447–451.
[34]
Thiery, J.; Keefe, D.; Saffarian, S.; Martinvalet, D.; Walch, M.; Boucrot, E.; Kirchhausen, T.; Lieberman, J. Perforin activates clathrin- and dynamin-dependent endocytosis, which is required for plasma membrane repair and delivery of granzyme B for granzyme-mediated apoptosis. Blood 2010, 115, 1582–1593.
[35]
Thiery, J.; Keefe, D.; Boulant, S.; Boucrot, E.; Walch, M.; Martinvalet, D.; Goping, I.S.; Bleackley, R.C.; Kirchhausen, T.; Lieberman, J. Perforin pores in the endosomal membrane trigger the release of endocytosed granzyme B into the cytosol of target cells. Nat. Immunol. 2011, 12, 770–777.
[36]
Gross, C.; Koelch, W.; DeMaio, A.; Arispe, N.; Multhoff, G. Cell surface-bound heat shock protein 70 (Hsp70) mediates perforin-independent apoptosis by specific binding and uptake of granzyme B. J. Biol. Chem. 2003, 278, 41173–41181.
[37]
Gehrmann, M.; Stangl, S.; Kirschner, A.; Foulds, G.A.; Sievert, W.; Doss, B.T.; Walch, A.; Pockley, A.G.; Multhoff, G. Immunotherapeutic targeting of membrane Hsp70-expressing tumors using recombinant human granzyme B. PLoS One 2012, 7, e41341.
Boivin, W.A.; Cooper, D.M.; Hiebert, P.R.; Granville, D.J. Intracellular versus extracellular granzyme B in immunity and disease: challenging the dogma. Lab. Invest. 2009, 89, 1195–1220, doi:10.1038/labinvest.2009.91.
[40]
Thornberry, N.A.; Rano, T.A.; Peterson, E.P.; Rasper, D.M.; Timkey, T.; Garcia-Calvo, M.; Houtzager, V.M.; Nordstrom, P.A.; Roy, S.; Vaillancourt, J.P.; et al. A combinatorial approach defines specificities of members of the caspase family and granzyme B. Functional relationships established for key mediators of apoptosis. J. Biol. Chem. 1997, 272, 17907–17911.
[41]
Adrain, C.; Murphy, B.M.; Martin, S.J. Molecular ordering of the caspase activation cascade initiated by the cytotoxic T lymphocyte/natural killer (CTL/NK) protease granzyme B. J. Biol. Chem. 2005, 280, 4663–4673, doi:10.1074/jbc.M410915200.
[42]
Barry, M.; Heibein, J.A.; Pinkoski, M.J.; Lee, S.F.; Moyer, R.W.; Green, D.R.; Bleackley, R.C. Granzyme B short-circuits the need for caspase 8 activity during granule-mediated cytotoxic T-lymphocyte killing by directly cleaving Bid. Mol. Cell. Biol. 2000, 20, 3781–3794, doi:10.1128/MCB.20.11.3781-3794.2000.
[43]
Waterhouse, N.J.; Sedelies, K.A.; Browne, K.A.; Wowk, M.E.; Newbold, A.; Sutton, V.R.; Clarke, C.J.; Oliaro, J.; Lindemann, R.K.; Bird, P.I.; et al. A central role for Bid in granzyme B-induced apoptosis. J. Biol. Chem. 2005, 280, 4476–4482.
[44]
Thomas, D.A.; Du, C.; Xu, M.; Wang, X.; Ley, T.J. DFF45/ICAD can be directly processed by granzyme B during the induction of apoptosis. Immunity 2000, 12, 621–632, doi:10.1016/S1074-7613(00)80213-7.
[45]
Sharif-Askari, E.; Alam, A.; Rheaume, E.; Beresford, P.J.; Scotto, C.; Sharma, K.; Lee, D.; DeWolf, W.E.; Nuttall, M.E.; Lieberman, J.; et al. Direct cleavage of the human DNA fragmentation factor-45 by granzyme B induces caspase-activated DNase release and DNA fragmentation. EMBO J. 2001, 20, 3101–3113.
[46]
Adrain, C.; Duriez, P.J.; Brumatti, G.; Delivani, P.; Martin, S.J. The cytotoxic lymphocyte protease, granzyme B, targets the cytoskeleton and perturbs microtubule polymerization dynamics. J. Biol. Chem. 2006, 281, 8118–8125.
[47]
Zhang, D.; Beresford, P.J.; Greenberg, A.H.; Lieberman, J. Granzymes A and B directly cleave lamins and disrupt the nuclear lamina during granule-mediated cytolysis. Proc. Natl. Acad. Sci. USA 2001, 98, 5746–5751.
[48]
Froelich, C.J.; Hanna, W.L.; Poirier, G.G.; Duriez, P.J.; D'Amours, D.; Salvesen, G.S.; Alnemri, E.S.; Earnshaw, W.C.; Shah, G.M. Granzyme B/perforin-mediated apoptosis of Jurkat cells results in cleavage of poly(ADP-ribose) polymerase to the 89-kDa apoptotic fragment and less abundant 64-kDa fragment. Biochem. Biophys. Res. Commun. 1996, 227, 658–665.
[49]
Giesübel, U.; D？lken, B.; Mahmud, H.; Wels, W.S. Cell binding, internalization and cytotoxic activity of human granzyme B expressed in the yeast Pichia pastoris. Biochem. J. 2006, 394, 563–573, doi:10.1042/BJ20050687.
[50]
Liu, Y.; Cheung, L.H.; Hittelman, W.N.; Rosenblum, M.G. Targeted delivery of human pro-apoptotic enzymes to tumor cells: In vitro studies describing a novel class of recombinant highly cytotoxic agents. Mol. Cancer Ther. 2003, 2, 1341–1350.
[51]
Kurschus, F.C.; Kleinschmidt, M.; Fellows, E.; Dornmair, K.; Rudolph, R.; Lilie, H.; Jenne, D.E. Killing of target cells by redirected granzyme B in the absence of perforin. FEBS Lett. 2004, 562, 87–92, doi:10.1016/S0014-5793(04)00187-5.
[52]
Lorentsen, R.H.; Fynbo, C.H.; Thogersen, H.C.; Etzerodt, M.; Holtet, T.L. Expression, refolding, and purification of recombinant human granzyme B. Protein Expr. Purif. 2005, 39, 18–26, doi:10.1016/j.pep.2004.08.017.
[53]
Stahnke, B.; Thepen, T.; Stocker, M.; Rosinke, R.; Jost, E.; Fischer, R.; Tur, M.K.; Barth, S. Granzyme B-H22(scFv), a human immunotoxin targeting CD64 in acute myeloid leukemia of monocytic subtypes. Mol. Cancer Ther. 2008, 7, 2924–2932, doi:10.1158/1535-7163.MCT-08-0554.
[54]
Gehrmann, M.; Doss, B.T.; Wagner, M.; Zettlitz, K.A.; Kontermann, R.E.; Foulds, G.; Pockley, A.G.; Multhoff, G. A novel expression and purification system for the production of enzymatic and biologically active human granzyme B. J. Immunol. Methods 2011, 371, 8–17, doi:10.1016/j.jim.2011.06.007.
[55]
Cereghino, J.L.; Cregg, J.M. Heterologous protein expression in the methylotrophic yeast Pichia pastoris. FEMS Microbiol. Rev. 2000, 24, 45–66, doi:10.1111/j.1574-6976.2000.tb00532.x.
[56]
Pham, C.T.; Thomas, D.A.; Mercer, J.D.; Ley, T.J. Production of fully active recombinant murine granzyme B in yeast. J. Biol. Chem. 1998, 273, 1629–1633.
[57]
Sun, J.; Bird, C.H.; Buzza, M.S.; McKee, K.E.; Whisstock, J.C.; Bird, P.I. Expression and purification of recombinant human granzyme B from Pichia pastoris. Biochem. Biophys. Res. Commun. 1999, 261, 251–255, doi:10.1006/bbrc.1999.0989.
[58]
D？lken, B.; Jabulowsky, R.A.; Oberoi, P.; Benhar, I.; Wels, W.S. Maltose-binding protein enhances secretion of recombinant human granzyme B accompanied by in vivo processing of a precursor MBP fusion protein. PLoS One 2010, 5, e14404.
Wels, W.; Harwerth, I.M.; Mueller, M.; Groner, B.; Hynes, N.E. Selective inhibition of tumor cell growth by a recombinant single-chain antibody-toxin specific for the erbB-2 receptor. Cancer Res. 1992, 52, 6310–6317.
[61]
Wels, W.; Beerli, R.; Hellmann, P.; Schmidt, M.; Marte, B.M.; Kornilova, E.S.; Hekele, A.; Mendelsohn, J.; Groner, B.; Hynes, N.E. EGF receptor and p185erbB-2-specific single-chain antibody toxins differ in their cell-killing activity on tumor cells expressing both receptor proteins. Int. J. Cancer 1995, 60, 137–144, doi:10.1002/ijc.2910600120.
[62]
Schmidt, M.; Wels, W. Targeted inhibition of tumour cell growth by a bispecific single-chain toxin containing an antibody domain and TGF alpha. Br. J. Cancer 1996, 74, 853–862, doi:10.1038/bjc.1996.448.
[63]
Zenke, M.; Steinlein, P.; Wagner, E.; Cotten, M.; Beug, H.; Birnstiel, M.L. Receptor-mediated endocytosis of transferrin-polycation conjugates: an efficient way to introduce DNA into hematopoietic cells. Proc. Natl. Acad. Sci. USA 1990, 87, 3655–3659.
[64]
Maurer-Gebhard, M.; Schmidt, M.; Azemar, M.; Altenschmidt, U.; St？cklin, E.; Wels, W.; Groner, B. Systemic treatment with a recombinant erbB-2 receptor-specific tumor toxin efficiently reduces pulmonary metastases in mice injected with genetically modified carcinoma cells. Cancer Res. 1998, 58, 2661–2666.
[65]
Kanatani, I.; Lin, X.; Yuan, X.; Manorek, G.; Shang, X.; Cheung, L.H.; Rosenblum, M.G.; Howell, S.B. Targeting granzyme B to tumor cells using a yoked human chorionic gonadotropin. Cancer Chemother. Pharmacol. 2011, 68, 979–990, doi:10.1007/s00280-011-1573-4.
[66]
Bird, C.H.; Sun, J.; Ung, K.; Karambalis, D.; Whisstock, J.C.; Trapani, J.A.; Bird, P.I. Cationic sites on granzyme B contribute to cytotoxicity by promoting its uptake into target cells. Mol. Cell. Biol. 2005, 25, 7854–7867, doi:10.1128/MCB.25.17.7854-7867.2005.
[67]
Shi, L.; Keefe, D.; Durand, E.; Feng, H.; Zhang, D.; Lieberman, J. Granzyme B binds to target cells mostly by charge and must be added at the same time as perforin to trigger apoptosis. J. Immunol. 2005, 174, 5456–5461.
[68]
Kurschus, F.C.; Fellows, E.; Stegmann, E.; Jenne, D.E. Granzyme B delivery via perforin is restricted by size, but not by heparan sulfate-dependent endocytosis. Proc. Natl. Acad. Sci. USA 2008, 105, 13799–13804.
Raja, S.M.; Metkar, S.S.; Honing, S.; Wang, B.; Russin, W.A.; Pipalia, N.H.; Menaa, C.; Belting, M.; Cao, X.; Dressel, R.; et al. A novel mechanism for protein delivery: Granzyme B undergoes electrostatic exchange from serglycin to target cells. J. Biol. Chem. 2005, 280, 20752–20761.
[71]
Kurschus, F.C.; Bruno, R.; Fellows, E.; Falk, C.S.; Jenne, D.E. Membrane receptors are not required to deliver granzyme B during killer cell attack. Blood 2005, 105, 2049–2058, doi:10.1182/blood-2004-06-2180.
[72]
Estebanez-Perpina, E.; Fuentes-Prior, P.; Belorgey, D.; Braun, M.; Kiefersauer, R.; Maskos, K.; Huber, R.; Rubin, H.; Bode, W. Crystal structure of the caspase activator human granzyme B, a proteinase highly specific for an Asp-P1 residue. Biol. Chem. 2000, 381, 1203–1214.
[73]
Buzza, M.S.; Zamurs, L.; Sun, J.; Bird, C.H.; Smith, A.I.; Trapani, J.A.; Froelich, C.J.; Nice, E.C.; Bird, P.I. Extracellular matrix remodeling by human granzyme B via cleavage of vitronectin, fibronectin, and laminin. J. Biol. Chem. 2005, 280, 23549–23558.
[74]
Tak, P.P.; Spaeny-Dekking, L.; Kraan, M.C.; Breedveld, F.C.; Froelich, C.J.; Hack, C.E. The levels of soluble granzyme A and B are elevated in plasma and synovial fluid of patients with rheumatoid arthritis (RA). Clin. Exp. Immunol. 1999, 116, 366–370, doi:10.1046/j.1365-2249.1999.00881.x.
[75]
Ronday, H.K.; van der Laan, W.H.; Tak, P.P.; de Roos, J.A.; Bank, R.A.; TeKoppele, J.M.; Froelich, C.J.; Hack, C.E.; Hogendoorn, P.C.; Breedveld, F.C.; et al. Human granzyme B mediates cartilage proteoglycan degradation and is expressed at the invasive front of the synovium in rheumatoid arthritis. Rheumatology (Oxford) 2001, 40, 55–61, doi:10.1093/rheumatology/40.1.55.
[76]
Skjelland, M.; Michelsen, A.E.; Krohg-Sorensen, K.; Tennoe, B.; Dahl, A.; Bakke, S.; Brosstad, F.; Damas, J.K.; Russell, D.; Halvorsen, B.; et al. lasma levels of granzyme B are increased in patients with lipid-rich carotid plaques as determined by echogenicity. Atherosclerosis 2007, 195, e142–146, doi:10.1016/j.atherosclerosis.2007.05.001.
[77]
Saito, Y.; Kondo, H.; Hojo, Y. Granzyme B as a novel factor involved in cardiovascular diseases. J. Cardiol. 2011, 57, 141–147, doi:10.1016/j.jjcc.2010.10.001.
[78]
Boivin, W.A.; Shackleford, M.; Vanden Hoek, A.; Zhao, H.; Hackett, T.L.; Knight, D.A.; Granville, D.J. Granzyme B cleaves decorin, biglycan and soluble betaglycan, releasing active transforming growth factor-beta1. PLoS One 2012, 7, e33163.
[79]
Wang, T.; Lee, M.H.; Choi, E.; Pardo-Villamizar, C.A.; Lee, S.B.; Yang, I.H.; Calabresi, P.A.; Nath, A. Granzyme B-induced neurotoxicity is mediated via activation of PAR-1 receptor and Kv1.3 channel. PLoS One 2012, 7, e43950.
Sun, J.; Bird, C.H.; Sutton, V.; McDonald, L.; Coughlin, P.B.; De Jong, T.A.; Trapani, J.A.; Bird, P.I. A cytosolic granzyme B inhibitor related to the viral apoptotic regulator cytokine response modifier A is present in cytotoxic lymphocytes. J. Biol. Chem. 1996, 271, 27802–27809.
[82]
Bird, C.H.; Sutton, V.R.; Sun, J.; Hirst, C.E.; Novak, A.; Kumar, S.; Trapani, J.A.; Bird, P.I. Selective regulation of apoptosis: The cytotoxic lymphocyte serpin proteinase inhibitor 9 protects against granzyme B-mediated apoptosis without perturbing the Fas cell death pathway. Mol. Cell. Biol. 1998, 18, 6387–6398.
[83]
Classen, C.F.; Bird, P.I.; Debatin, K.M. Modulation of the granzyme B inhibitor proteinase inhibitor 9 (PI-9) by activation of lymphocytes and monocytes in vitro and by Epstein-Barr virus and bacterial infection. Clin. Exp. Immunol. 2006, 143, 534–542, doi:10.1111/j.1365-2249.2006.03006.x.
[84]
Hirst, C.E.; Buzza, M.S.; Bird, C.H.; Warren, H.S.; Cameron, P.U.; Zhang, M.; Ashton-Rickardt, P.G.; Bird, P.I. The intracellular granzyme B inhibitor, proteinase inhibitor 9, is up-regulated during accessory cell maturation and effector cell degranulation, and its overexpression enhances CTL potency. J. Immunol. 2003, 170, 805–815.
[85]
Bots, M.; de Bruin, E.; Rademaker-Koot, M.T.; Medema, J.P. Proteinase inhibitor-9 expression is induced by maturation in dendritic cells via p38 MAP kinase. Hum. Immunol. 2007, 68, 959–964, doi:10.1016/j.humimm.2007.10.011.
[86]
Medema, J.P.; de Jong, J.; Peltenburg, L.T.; Verdegaal, E.M.; Gorter, A.; Bres, S.A.; Franken, K.L.; Hahne, M.; Albar, J.P.; Melief, C.J.; et al. Blockade of the granzyme B/perforin pathway through overexpression of the serine protease inhibitor PI-9/SPI-6 constitutes a mechanism for immune escape by tumors. Proc. Natl. Acad. Sci. USA 2001, 98, 11515–11520.
[87]
Rousalova, I.; Krepela, E.; Prochazka, J.; Cermak, J.; Benkova, K. Expression of proteinase inhibitor-9/serpinB9 in non-small cell lung carcinoma cells and tissues. Int. J. Oncol. 2010, 36, 275–283.
[88]
ten Berge, R.L.; Meijer, C.J.; Dukers, D.F.; Kummer, J.A.; Bladergroen, B.A.; Vos, W.; Hack, C.E.; Ossenkoppele, G.J.; Oudejans, J.J. Expression levels of apoptosis-related proteins predict clinical outcome in anaplastic large cell lymphoma. Blood 2002, 99, 4540–4546, doi:10.1182/blood.V99.12.4540.
[89]
van Houdt, I.S.; Oudejans, J.J.; van den Eertwegh, A.J.; Baars, A.; Vos, W.; Bladergroen, B.A.; Rimoldi, D.; Muris, J.J.; Hooijberg, E.; Gundy, C.M.; et al. Expression of the apoptosis inhibitor protease inhibitor 9 predicts clinical outcome in vaccinated patients with stage III and IV melanoma. Clin. Cancer Res. 2005, 11, 6400–6407.
[90]
Soriano, C.; Mukaro, V.; Hodge, G.; Ahern, J.; Holmes, M.; Jersmann, H.; Moffat, D.; Meredith, D.; Jurisevic, C.; Reynolds, P.N.; et al. Increased proteinase inhibitor-9 (PI-9) and reduced granzyme B in lung cancer: mechanism for immune evasion? Lung Cancer 2012, 77, 38–45, doi:10.1016/j.lungcan.2012.01.017.
[91]
Losasso, V.; Schiffer, S.; Barth, S.; Carloni, P. Design of human granzyme B variants resistant to serpin B9. Proteins 2012, 80, 2514–2522.
[92]
Sutton, V.R.; Sedelies, K.; Dewson, G.; Christensen, M.E.; Bird, P.I.; Johnstone, R.W.; Kluck, R.M.; Trapani, J.A.; Waterhouse, N.J. Granzyme B triggers a prolonged pressure to die in Bcl-2 overexpressing cells, defining a window of opportunity for effective treatment with ABT-737. Cell Death Dis. 2012, 3, e344, doi:10.1038/cddis.2012.73.
[93]
Zhang, X.; Sawyer, G.J.; Dong, X.; Qiu, Y.; Collins, L.; Fabre, J.W. The in vivo use of chloroquine to promote non-viral gene delivery to the liver via the portal vein and bile duct. J. Gene Med. 2003, 5, 209–218, doi:10.1002/jgm.340.
[94]
Raynes, K. Bisquinoline antimalarials: their role in malaria chemotherapy. Int. J. Parasitol. 1999, 29, 367–379, doi:10.1016/S0020-7519(98)00217-3.
[95]
Zhao, J.; Zhang, L.H.; Jia, L.T.; Zhang, L.; Xu, Y.M.; Wang, Z.; Yu, C.J.; Peng, W.D.; Wen, W.H.; Wang, C.J.; et al. Secreted antibody/granzyme B fusion protein stimulates selective killing of HER2-overexpressing tumor cells. J. Biol. Chem. 2004, 279, 21343–21348.
[96]
Fominaya, J.; Wels, W. Target cell-specific DNA transfer mediated by a chimeric multidomain protein. Novel non-viral gene delivery system. J. Biol. Chem. 1996, 271, 10560–10568, doi:10.1074/jbc.271.18.10560.
[97]
Uherek, C.; Fominaya, J.; Wels, W. A modular DNA carrier protein based on the structure of diphtheria toxin mediates target cell-specific gene delivery. J. Biol. Chem. 1998, 273, 8835–8841, doi:10.1074/jbc.273.15.8835.
[98]
Antignani, A.; Youle, R.J. How do Bax and Bak lead to permeabilization of the outer mitochondrial membrane? Curr. Opin. Cell Biol. 2006, 18, 685–689, doi:10.1016/j.ceb.2006.10.004.
[99]
Sampieri, K.; Fodde, R. Cancer stem cells and metastasis. Semin. Cancer Biol. 2012, 22, 187–193.
[100]
Robertson, M.J.; Cochran, K.J.; Cameron, C.; Le, J.M.; Tantravahi, R.; Ritz, J. Characterization of a cell line, NKL, derived from an aggressive human natural killer cell leukemia. Exp. Hematol. 1996, 24, 406–415.
