Genetically Engineered Mouse Models of Pancreatic Cancer: The KPC Model (LSL‐KrasG12D/+;LSL‐Trp53R172H/+;Pdx‐1‐Cre), Its Variants, and Their Application in Immuno‐oncology Drug Discovery

Jae W. Lee1, Chad A. Komar1, Fee Bengsch1, Kathleen Graham1, Gregory L. Beatty1

1 Division of Hematology‐Oncology, Department of Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania
Publication Name:  Current Protocols in Pharmacology
Unit Number:  Unit 14.39
DOI:  10.1002/cpph.2
Online Posting Date:  June, 2016
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Abstract

Pancreatic ductal adenocarcinoma (PDAC) ranks fourth among cancer‐related deaths in the United States. For patients with unresectable disease, treatment options are limited and lack curative potential. Preclinical mouse models of PDAC that recapitulate the biology of human pancreatic cancer offer an opportunity for the rational development of novel treatment approaches that may improve patient outcomes. With the recent success of immunotherapy for subsets of patients with solid malignancies, interest is mounting in the possible use of immunotherapy for the treatment of PDAC. Considered in this unit is the value of genetic mouse models for characterizing the immunobiology of PDAC and for investigating novel immunotherapeutics. Several variants of these models are described, all of which may be used in drug development and for providing information on unique aspects of disease biology and therapeutic responsiveness. © 2016 by John Wiley & Sons, Inc.

Keywords: genetically engineered mouse model; immunotherapy; inflammation; KPC; macrophage; pancreatic ductal adenocarcinoma; T cell exclusion

     
 
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Table of Contents

  • Introduction
  • Genetic Mouse Models of PDAC
  • Immunobiology of Human and Mouse PDAC
  • Immuno‐Oncology Discovery Using Genetic Models of PDAC
  • The Value of Genetic Models for Evaluating Immunotherapy In PDAC
  • Application of Genetic Models of PDAC and Their Variants to Preclinical Development of Immunotherapy
  • Conclusions
  • Literature Cited
  • Figures
     
 
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Materials

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Literature Cited

Literature Cited
  Ansell, S.M., Lesokhin, A.M., Borrello, I., Halwani, A., Scott, E.C., Gutierrez, M., Schuster, S.J., Millenson, M.M., Cattry, D., Freeman, G.J., Rodig, S.J., Chapuy, B., Ligon, A.H., Zhu, L., Grosso, J.F., Kim, S.Y., Timmerman, J.M., Shipp, M.A., and Armand, P. 2015. PD‐1 blockade with nivolumab in relapsed or refractory Hodgkin's lymphoma. N. Engl. J. Med. 372:311‐319. doi: 10.1056/NEJMoa1411087.
  Barugola, G., Partelli, S., Marcucci, S., Sartori, N., Capelli, P., Bassi, C., Pederzoli, P., and Falconi, M. 2009. Resectable pancreatic cancer: Who really benefits from resection? Ann. Surg. Oncol. 16:3316‐3322. doi: 10.1245/s10434‐009‐0670‐7.
  Bayne, L.J., Beatty, G.L., Jhala, N., Clark, C.E., Rhim, A.D., Stanger, B.Z., and Vonderheide, R.H. 2012. Tumor‐derived granulocyte‐macrophage colony‐stimulating factor regulates myeloid inflammation and T cell immunity in pancreatic cancer. Cancer Cell 21:822‐835. doi: 10.1016/j.ccr.2012.04.025.
  Beatty, G.L. and Gladney, W.L. 2015. Immune escape mechanisms as a guide for cancer immunotherapy. Clin. Cancer Res. 21:687‐692. doi: 10.1158/1078‐0432.CCR‐14‐1860.
  Beatty, G.L. and Paterson, Y. 2000. IFN‐gamma can promote tumor evasion of the immune system in vivo by down‐regulating cellular levels of an endogenous tumor antigen. J. Immunol. 165:5502‐5508. doi: 10.4049/jimmunol.165.10.5502.
  Beatty, G.L., Chiorean, E.G., Fishman, M.P., Saboury, B., Teitelbaum, U.R., Sun, W., Huhn, R.D., Song, W., Li, D., Sharp, L.L., Torigian, D.A., O'Dwyer, P.J., and Vonderheide, R.H. 2011. CD40 agonists alter tumor stroma and show efficacy against pancreatic carcinoma in mice and humans. Science 331:1612‐1616. doi: 10.1126/science.1198443.
  Beatty, G.L., Torigian, D.A., Chiorean, E.G., Saboury, B., Brothers, A., Alavi, A., Troxel, A.B., Sun, W., Teitelbaum, U.R., Vonderheide, R.H., and O'Dwyer, P. 2013. A phase I study of an agonist CD40 monoclonal antibody (CP‐870,893) in combination with gemcitabine in patients with advanced pancreatic ductal adenocarcinoma. Clin. Cancer Res. 19:6286‐6295. doi: 10.1158/1078‐0432.CCR‐13‐1320.
  Beatty, G.L., Winograd, R., Evans, R.A., Long, K.B., Luque, S.L., Lee, J.W., Clendenin, C., Gladney, W.L., Knoblock, D.M., Guirnalda, P.D., and Vonderheide, R.H. 2015. Exclusion of T cells from pancreatic carcinomas in mice is regulated by Ly6C(low) F4/80(+) extratumoral macrophages. Gastroenterology 149:201‐210. doi: 10.1053/j.gastro.2015.04.010.
  Biankin, A.V., Waddell, N., Kassahn, K.S., Gingras, M.C., Muthuswamy, L.B., Johns, A.L., Miller, D.K., Wilson, P.J., Patch, A.M., Wu, J., Chang, D.K., Cowley, M.J., Gardiner, B.B., Song, S., Harliwong, I., Idrisoglu, S., Nourse, C., Nourbakhsh, E., Manning, S., Wani, S., Gongora, M., Pajic, M., Scarlett, C.J., Gill, A.J., Pinho, A.V., Rooman, I., Anderson, M., Holmes, O., Leonard, C., Taylor, D., Wood, S., Xu, Q., Nones, K., Fink, J.L., Christ, A., Bruxner, T., Cloonan, N., Kolle, G., Newell, F., Pinese, M., Mead, R.S., Humphris, J.L., Kaplan, W., Jones, M.D., Colvin, E.K., Nagrial, A.M., Humphrey, E.S., Chou, A., Chin, V.T., Chantrill, L.A., Mawson, A., Samra, J.S., Kench, J.G., Lovell, J.A., Daly, R.J., Merrett, N.D., Toon, C., Epari, K., Nguyen, N.Q., Barbour, A., Zeps, N., Kakkar, N., Zhao, F., Wu, Y.Q., Wang, M., Muzny, D.M., Fisher, W.E., Brunicardi, F.C., Hodges, S.E., Reid, J.G., Drummond, J., Chang, K., Han, Y., Lewis, L.R., Dinh, H., Buhay, C.J., Beck, T., Timms, L., Sam, M., Begley, K., Brown, A., Pai, D., Panchal, A., Buchner, N., De Borja, R., Denroche, R.E., Yung, C.K., Serra, S., Onetto, N., Mukhopadhyay, D., Tsao, M.S., Shaw, P.A., Petersen, G.M., Gallinger, S., Hruban, R.H., Maitra, A., Iacobuzio‐Donahue, C.A., Schulick, R.D., Wolfgang, C.L., Morgan, R.A., Lawlor, R.T., Capelli, P., Corbo, V., Scardoni, M., Tortora, G., Tempero, M.A., Mann, K.M., Jenkins, N.A., Perez‐Mancera, P.A., Adams, D.J., Largaespada, D.A., Wessels, L.F., Rust, A.G., Stein, L.D., Tuveson, D.A., Copeland, N.G., Musgrove, E.A., Scarpa, A., Eshleman, J.R., Hudson, T.J., Sutherland, R.L., Wheeler, D.A., Pearson, J.V., McPherson, J.D., Gibbs, R.A., and Grimmond, S.M. 2012. Pancreatic cancer genomes reveal aberrations in axon guidance pathway genes. Nature 491:399‐405. doi: 10.1038/nature11547.
  Boj, S.F., Hwang, C.I., Baker, L.A., Chio, I.I., Engle, D.D., Corbo, V., Jager, M., Ponz‐Sarvise, M., Tiriac, H., Spector, M.S., Gracanin, A., Oni, T., Yu, K.H., van Boxtel, R., Huch, M., Rivera, K.D., Wilson, J.P., Feigin, M.E., Ohlund, D., Handly‐Santana, A., Ardito‐Abraham, C.M., Ludwig, M., Elyada, E., Alagesan, B., Biffi, G., Yordanov, G.N., Delcuze, B., Creighton, B., Wright, K., Park, Y., Morsink, F.H., Molenaar, I.Q., BorelRinkes, I.H., Cuppen, E., Hao, Y., Jin, Y., Nijman, I.J., Iacobuzio‐Donahue, C., Leach, S.D., Pappin, D.J., Hammell, M., Klimstra, D.S., Basturk, O., Hruban, R.H., Offerhaus, G.J., Vries, R.G., Clevers, H., and Tuveson, D.A. 2015. Organoid models of human and mouse ductal pancreatic cancer. Cell 160:324‐338. doi: 10.1016/j.cell.2014.12.021.
  Brahmer, J.R., Tykodi, S.S., Chow, L.Q., Hwu, W.J., Topalian, S.L., Hwu, P., Drake, C.G., Camacho, L.H., Kauh, J., Odunsi, K., Pitot, H.C., Hamid, O., Bhatia, S., Martins, R., Eaton, K., Chen, S., Salay, T.M., Alaparthy, S., Grosso, J.F., Korman, A.J., Parker, S.M., Agrawal, S., Goldberg, S.M., Pardoll, D.M., Gupta, A., and Wigginton, J.M. 2012. Safety and activity of anti‐PD‐L1 antibody in patients with advanced cancer. N. Engl. J. Med. 366:2455‐2465. doi: 10.1056/NEJMoa1200694.
  Chang, D.Z., Ma, Y., Ji, B., Wang, H., Deng, D., Liu, Y., Abbruzzese, J.L., Liu, Y.J., Logsdon, C.D., and Hwu, P. 2011. Mast cells in tumor microenvironment promotes the in vivo growth of pancreatic ductal adenocarcinoma. Clin. Cancer Res. 17:7015‐7023. doi: 10.1158/1078‐0432.CCR‐11‐0607.
  Colvin, E.K. and Scarlett, C.J. 2014. A historical perspective of pancreatic cancer mouse models. Semin. Cell Dev. Biol. 27:96‐105. doi: 10.1016/j.semcdb.2014.03.025.
  Costa‐Silva, B., Aiello, N.M., Ocean, A.J., Singh, S., Zhang, H., Thakur, B.K., Becker, A., Hoshino, A., Mark, M.T., Molina, H., Xiang, J., Zhang, T., Theilen, T.M., Garcia‐Santos, G., Williams, C., Ararso, Y., Huang, Y., Rodrigues, G., Shen, T.L., Labori, K.J., Lothe, I.M., Kure, E.H., Hernandez, J., Doussot, A., Ebbesen, S.H., Grandgenett, P.M., Hollingsworth, M.A., Jain, M., Mallya, K., Batra, S.K., Jarnagin, W.R., Schwartz, R.E., Matei, I., Peinado, H., Stanger, B.Z., Bromberg, J., and Lyden, D. 2015. Pancreatic cancer exosomes initiate pre‐metastatic niche formation in the liver. Nat. Cell Biol. 17:816‐826. doi: 10.1038/ncb3169.
  De Monte, L., Reni, M., Tassi, E., Clavenna, D., Papa, I., Recalde, H., Braga, M., Di Carlo, V., Doglioni, C., and Protti, M.P. 2011. Intratumor T helper type 2 cell infiltrate correlates with cancer‐associated fibroblast thymic stromal lymphopoietin production and reduced survival in pancreatic cancer. J. Exp. Med. 208:469‐478. doi: 10.1084/jem.20101876.
  DeCant, B.T., Principe, D.R., Guerra, C., Pasca di Magliano, M., and Grippo, P.J. 2014. Utilizing past and present mouse systems to engineer more relevant pancreatic cancer models. Front. Physiol. 5:464. doi: 10.3389/fphys.2014.00464.
  Diehl, L., den Boer, A.T., Schoenberger, S.P., van der Voort, E.I., Schumacher, T.N., Melief, C.J., Offringa, R., and Toes, R.E. 1999. CD40 activation in vivo overcomes peptide‐induced peripheral cytotoxic T‐lymphocyte tolerance and augments anti‐tumor vaccine efficacy. Nat. Med. 5:774‐779. doi: 10.1038/10495.
  Ding, Y., Cravero, J.D., Adrian, K., and Grippo, P. 2010. Modeling pancreatic cancer in vivo: From xenograft and carcinogen‐induced systems to genetically engineered mice. Pancreas 39:283‐292. doi: 10.1097/MPA.0b013e3181c15619.
  Faca, V.M., Song, K.S., Wang, H., Zhang, Q., Krasnoselsky, A.L., Newcomb, L.F., Plentz, R.R., Gurumurthy, S., Redston, M.S., Pitteri, S.J., Pereira‐Faca, S.R., Ireton, R.C., Katayama, H., Glukhova, V., Phanstiel, D., Brenner, D.E., Anderson, M.A., Misek, D., Scholler, N., Urban, N.D., Barnett, M.J., Edelstein, C., Goodman, G.E., Thornquist, M.D., McIntosh, M.W., DePinho, R.A., Bardeesy, N., and Hanash, S.M. 2008. A mouse to human search for plasma proteome changes associated with pancreatic tumor development. PLoS Med. 5:e123. doi: 10.1371/journal.pmed.0050123.
  Feig, C., Jones, J.O., Kraman, M., Wells, R.J., Deonarine, A., Chan, D.S., Connell, C.M., Roberts, E.W., Zhao, Q., Caballero, O.L., Teichmann, S.A., Janowitz, T., Jodrell, D.I., Tuveson, D.A., and Fearon, D.T. 2013. Targeting CXCL12 from FAP‐expressing carcinoma‐associated fibroblasts synergizes with anti‐PD‐L1 immunotherapy in pancreatic cancer. Proc. Natl. Acad. Sci. U. S. A. 110:20212‐20217. doi: 10.1073/pnas.1320318110.
  French, R.R., Chan, H.T., Tutt, A.L., and Glennie, M.J. 1999. CD40 antibody evokes a cytotoxic T‐cell response that eradicates lymphoma and bypasses T‐cell help. Nat. Med. 5:548‐553. doi: 10.1038/5505.
  Fukuda, A., Wang, S.C., Morris, J.P. 4th, Folias, A.E., Liou, A., Kim, G.E., Akira, S., Boucher, K.M., Firpo, M.A., Mulvihill, S.J., and Hebrok, M. 2011. Stat3 and MMP7 contribute to pancreatic ductal adenocarcinoma initiation and progression. Cancer Cell 19:441‐455. doi: 10.1016/j.ccr.2011.03.002.
  Fukunaga, A., Miyamoto, M., Cho, Y., Murakami, S., Kawarada, Y., Oshikiri, T., Kato, K., Kurokawa, T., Suzuoki, M., Nakakubo, Y., Hiraoka, K., Itoh, T., Morikawa, T., Okushiba, S., Kondo, S., and Katoh, H. 2004. CD8+ tumor‐infiltrating lymphocytes together with CD4+ tumor‐infiltrating lymphocytes and dendritic cells improve the prognosis of patients with pancreatic adenocarcinoma. Pancreas 28:e26‐e31. doi: 10.1097/00006676‐200401000‐00023.
  Galon, J., Costes, A., Sanchez‐Cabo, F., Kirilovsky, A., Mlecnik, B., Lagorce‐Pages, C., Tosolini, M., Camus, M., Berger, A., Wind, P., Zinzindohoue, F., Bruneval, P., Cugnenc, P.H., Trajanoski, Z., Fridman, W.H., and Pages, F. 2006. Type, density, and location of immune cells within human colorectal tumors predict clinical outcome. Science 313:1960‐1964. doi: 10.1126/science.1129139.
  Garon, E.B., Rizvi, N.A., Hui, R., Leighl, N., Balmanoukian, A.S., Eder, J.P., Patnaik, A., Aggarwal, C., Gubens, M., Horn, L., Carcereny, E., Ahn, M.J., Felip, E., Lee, J.S., Hellmann, M.D., Hamid, O., Goldman, J.W., Soria, J.C., Dolled‐Filhart, M., Rutledge, R.Z., Zhang, J., Lunceford, J.K., Rangwala, R., Lubiniecki, G.M., Roach, C., Emancipator, K., and Gandhi, L. 2015. Pembrolizumab for the treatment of non‐small‐cell lung cancer. N. Engl. J. Med. 372:2018‐2028. doi: 10.1056/NEJMoa1501824.
  Gubin, M.M., Zhang, X., Schuster, H., Caron, E., Ward, J.P., Noguchi, T., Ivanova, Y., Hundal, J., Arthur, C.D., Krebber, W.J., Mulder, G.E., Toebes, M., Vesely, M.D., Lam, S.S., Korman, A.J., Allison, J.P., Freeman, G.J., Sharpe, A.H., Pearce, E.L., Schumacher, T.N., Aebersold, R., Rammensee, H.G., Melief, C.J., Mardis, E.R., Gillanders, W.E., Artyomov, M.N., and Schreiber, R.D. 2014. Checkpoint blockade cancer immunotherapy targets tumour‐specific mutant antigens. Nature 515:577‐581. doi: 10.1038/nature13988.
  Hidalgo, M., Amant, F., Biankin, A.V., Budinska, E., Byrne, A.T., Caldas, C., Clarke, R.B., de Jong, S., Jonkers, J., Maelandsmo, G.M., Roman‐Roman, S., Seoane, J., Trusolino, L., and Villanueva, A. 2014. Patient‐derived xenograft models: An emerging platform for translational cancer research. Cancer Discov. 4:998‐1013. doi: 10.1158/2159‐8290.CD‐14‐0001.
  Hingorani, S.R., Petricoin, E.F., Maitra, A., Rajapakse, V., King, C., Jacobetz, M.A., Ross, S., Conrads, T.P., Veenstra, T.D., Hitt, B.A., Kawaguchi, Y., Johann, D., Liotta, L.A., Crawford, H.C., Putt, M.E., Jacks, T., Wright, C.V., Hruban, R.H., Lowy, A.M., and Tuveson, D.A. 2003. Preinvasive and invasive ductal pancreatic cancer and its early detection in the mouse. Cancer Cell 4:437‐450. doi: 10.1016/S1535‐6108(03) 00309‐X.
  Hingorani, S.R., Wang, L., Multani, A.S., Combs, C., Deramaudt, T.B., Hruban, R.H., Rustgi, A.K., Chang, S., and Tuveson, D.A. 2005. Trp53R172H and KrasG12D cooperate to promote chromosomal instability and widely metastatic pancreatic ductal adenocarcinoma in mice. Cancer Cell 7:469‐483. doi: 10.1016/j.ccr.2005.04.023.
  Hodi, F.S., O'Day, S.J., McDermott, D.F., Weber, R.W., Sosman, J.A., Haanen, J.B., Gonzalez, R., Robert, C., Schadendorf, D., Hassel, J.C., Akerley, W., van den Eertwegh, A.J., Lutzky, J., Lorigan, P., Vaubel, J.M., Linette, G.P., Hogg, D., Ottensmeier, C.H., Lebbe, C., Peschel, C., Quirt, I., Clark, J.I., Wolchok, J.D., Weber, J.S., Tian, J., Yellin, M.J., Nichol, G.M., Hoos, A., and Urba, W.J. 2010. Improved survival with ipilimumab in patients with metastatic melanoma. N. Engl. J. Med. 363:711‐723. doi: 10.1056/NEJMoa1003466.
  Hruban, R.H., Adsay, N.V., Albores‐Saavedra, J., Compton, C., Garrett, E.S., Goodman, S.N., Kern, S.E., Klimstra, D.S., Kloppel, G., Longnecker, D.S., Luttges, J., and Offerhaus, G.J. 2001. Pancreatic intraepithelial neoplasia: A new nomenclature and classification system for pancreatic duct lesions. Am. J. Surg. Pathol. 25:579‐586. doi: 10.1097/00000478‐200105000‐00003.
  Hwang, C.I., Boj, S.F., Clevers, H., and Tuveson, D.A. 2015. Preclinical models of pancreatic ductal adenocarcinoma. J. Pathol. 238:197‐204. doi: 10.1002/path.4651.
  Ino, Y., Yamazaki‐Itoh, R., Shimada, K., Iwasaki, M., Kosuge, T., Kanai, Y., and Hiraoka, N. 2013. Immune cell infiltration as an indicator of the immune microenvironment of pancreatic cancer. Br. J. Cancer 108:914‐923. doi: 10.1038/bjc.2013.32.
  Jacobetz, M.A., Chan, D.S., Neesse, A., Bapiro, T.E., Cook, N., Frese, K.K., Feig, C., Nakagawa, T., Caldwell, M.E., Zecchini, H.I., Lolkema, M.P., Jiang, P., Kultti, A., Thompson, C.B., Maneval, D.C., Jodrell, D.I., Frost, G.I., Shepard, H.M., Skepper, J.N., and Tuveson, D.A. 2013. Hyaluronan impairs vascular function and drug delivery in a mouse model of pancreatic cancer. Gut 62:112‐120. doi: 10.1136/gutjnl‐2012‐302529.
  Jones, S., Zhang, X., Parsons, D.W., Lin, J.C., Leary, R.J., Angenendt, P., Mankoo, P., Carter, H., Kamiyama, H., Jimeno, A., Hong, S.M., Fu, B., Lin, M.T., Calhoun, E.S., Kamiyama, M., Walter, K., Nikolskaya, T., Nikolsky, Y., Hartigan, J., Smith, D.R., Hidalgo, M., Leach, S.D., Klein, A.P., Jaffee, E.M., Goggins, M., Maitra, A., Iacobuzio‐Donahue, C., Eshleman, J.R., Kern, S.E., Hruban, R.H., Karchin, R., Papadopoulos, N., Parmigiani, G., Vogelstein, B., Velculescu, V.E., and Kinzler, K.W. 2008. Core signaling pathways in human pancreatic cancers revealed by global genomic analyses. Science 321:1801‐1806. doi: 10.1126/science.1164368.
  Josefowicz, S.Z., Lu, L.F., and Rudensky, A.Y. 2012. Regulatory T cells: Mechanisms of differentiation and function. Annu. Rev. Immunol. 30:531‐564. doi: 10.1146/annurev.immunol.25.022106.141623.
  Joyce, J.A. and Fearon, D.T. 2015. T cell exclusion, immune privilege, and the tumor microenvironment. Science 348:74‐80. doi: 10.1126/science.aaa6204.
  Keenan, B.P., Saenger, Y., Kafrouni, M.I., Leubner, A., Lauer, P., Maitra, A., Rucki, A.A., Gunderson, A.J., Coussens, L.M., Brockstedt, D.G., Dubensky, T.W., Jr., Hassan, R., Armstrong, T.D., and Jaffee, E.M. 2014. A Listeria vaccine and depletion of T‐regulatory cells activate immunity against early stage pancreatic intraepithelial neoplasms and prolong survival of mice. Gastroenterology 146:1784‐1794. doi: 10.1053/j.gastro.2014.02.055.
  Le, D.T., Lutz, E., Uram, J.N., Sugar, E.A., Onners, B., Solt, S., Zheng, L., Diaz, L.A., Jr., Donehower, R.C., Jaffee, E.M., and Laheru, D.A. 2013. Evaluation of ipilimumab in combination with allogeneic pancreatic tumor cells transfected with a GM‐CSF gene in previously treated pancreatic cancer. J. Immunother. 36:382‐389. doi: 10.1097/CJI.0b013e31829fb7a2.
  Le, D.T., Uram, J.N., Wang, H., Bartlett, B.R., Kemberling, H., Eyring, A.D., Skora, A.D., Luber, B.S., Azad, N.S., Laheru, D., Biedrzycki, B.,Donehower, R.C., Zaheer, A., Fisher, G.A., Crocenzi, T.S., Lee, J.J., Duffy, S.M., Goldberg, R.M., de la Chapelle, A., Koshiji, M., Bhaijee, F., Huebner, T., Hruban, R.H., Wood, L.D., Cuka, N., Pardoll, D.M., Papadopoulos, N., Kinzler, K.W., Zhou, S., Cornish, T.C., Taube, J.M., Anders, R.A., Eshleman, J.R., Vogelstein, B., and Diaz, L.A., Jr. 2015a. PD‐1 blockade in tumors with mismatch‐repair deficiency. N. Engl. J. Med. 372:2509‐2520. doi: 10.1056/NEJMoa1500596.
  Le, D.T., Wang‐Gillam, A., Picozzi, V., Greten, T.F., Crocenzi, T., Springett, G., Morse, M., Zeh, H., Cohen, D., Fine, R.L., Onners, B., Uram, J.N., Laheru, D.A., Lutz, E.R., Solt, S., Murphy, A.L., Skoble, J., Lemmens, E., Grous, J., Dubensky, T., Jr., Brockstedt, D.G., and Jaffee, E.M. 2015b. Safety and survival with GVAX pancreas prime and listeria monocytogenes‐expressing mesothelin (CRS‐207) boost vaccines for metastatic pancreatic cancer. J. Clin. Oncol. 33:1325‐1333. doi: 10.1200/JCO.2014.57.4244.
  Lesina, M., Kurkowski, M.U., Ludes, K., Rose‐John, S., Treiber, M., Kloppel, G., Yoshimura, A., Reindl, W., Sipos, B., Akira, S., Schmid, R.M., and Algul, H. 2011. Stat3/Socs3 activation by IL‐6 transsignaling promotes progression of pancreatic intraepithelial neoplasia and development of pancreatic cancer. Cancer Cell 19:456‐469. doi: 10.1016/j.ccr.2011.03.009.
  Lesokhin, A.M., Callahan, M.K., Postow, M.A., and Wolchok, J.D. 2015. On being less tolerant: Enhanced cancer immunosurveillance enabled by targeting checkpoints and agonists of T cell activation. Sci. Transl. Med. 7:280sr281. doi: 10.1126/scitranslmed.3010274.
  Liou, G.Y., Doppler, H., Necela, B., Edenfield, B., Zhang, L., Dawson, D.W., and Storz, P. 2015. Mutant KRAS‐induced expression of ICAM‐1 in pancreatic acinar cells causes attraction of macrophages to expedite the formation of precancerous lesions. Cancer Discov. 5:52‐63. doi: 10.1158/2159‐8290.CD‐14‐0474.
  Lo, A., Wang, L.C., Scholler, J., Monslow, J., Avery, D., Newick, K., O'Brien, S., Evans, R.A., Bajor, D.L., Clendenin, C., Durham, A.C., Buza, E.L., Vonderheide, R.H., June, C.H., Albelda, S.M., and Pure, E. 2015. Tumor‐promoting desmoplasia is disrupted by depleting FAP‐expressing stromal cells. Cancer Res. 75:2800‐2810. doi: 10.1158/0008‐5472.CAN‐14‐3041.
  Long, K.B. and Beatty, G.L. 2013. Harnessing the antitumor potential of macrophages for cancer immunotherapy. Oncoimmunology 2:e26860. doi: 10.4161/onci.26860.
  Lutz, E.R., Wu, A.A., Bigelow, E., Sharma, R., Mo, G., Soares, K., Solt, S., Dorman, A., Wamwea, A., Yager, A., Laheru, D., Wolfgang, C.L., Wang, J., Hruban, R.H., Anders, R.A., Jaffee, E.M., and Zheng, L. 2014. Immunotherapy converts nonimmunogenic pancreatic tumors into immunogenic foci of immune regulation. Cancer Immunol. Res. 2:616‐631. doi: 10.1158/2326‐6066.CIR‐14‐0027.
  Maddipati, R. and Stanger, B.Z. 2015. Pancreatic cancer metastases harbor evidence of polyclonality. Cancer Discov. 5:1086‐1097. doi: 10.1158/2159‐8290.CD‐15‐0120.
  Mann, O., Strate, T., Schneider, C., Yekebas, E.F., and Izbicki, J.R. 2006. Surgery for advanced and metastatic pancreatic cancer‐current state and perspectives. Anticancer Res. 26:681‐686.
  Matsushita, H., Vesely, M.D., Koboldt, D.C., Rickert, C.G., Uppaluri, R., Magrini, V.J., Arthur, C.D., White, J.M., Chen, Y.S., Shea, L.K., Hundal, J., Wendl, M.C., Demeter, R., Wylie, T., Allison, J.P., Smyth, M.J., Old, L.J., Mardis, E.R., and Schreiber, R.D. 2012. Cancer exome analysis reveals a T‐cell‐dependent mechanism of cancer immunoediting. Nature 482:400‐404. doi: 10.1038/nature10755.
  McAllister, F., Bailey, J.M., Alsina, J., Nirschl, C.J., Sharma, R., Fan, H., Rattigan, Y., Roeser, J.C., Lankapalli, R.H., Zhang, H., Jaffee, E.M., Drake, C.G., Housseau, F., Maitra, A., Kolls, J.K., Sears, C.L., Pardoll, D.M., and Leach, S.D. 2014. Oncogenic Kras activates a hematopoietic‐to‐epithelial IL‐17 signaling axis in preinvasive pancreatic neoplasia. Cancer Cell 25:621‐637. doi: 10.1016/j.ccr.2014.03.014.
  Melo, S.A., Luecke, L.B., Kahlert, C., Fernandez, A.F., Gammon, S.T., Kaye, J., LeBleu, V.S., Mittendorf, E.A., Weitz, J., Rahbari, N., Reissfelder, C., Pilarsky, C., Fraga, M.F., Piwnica‐Worms, D., and Kalluri, R. 2015. Glypican‐1 identifies cancer exosomes and detects early pancreatic cancer. Nature 523:177‐182. doi: 10.1038/nature14581.
  Mitchem, J.B., Brennan, D.J., Knolhoff, B.L., Belt, B.A., Zhu, Y., Sanford, D.E., Belaygorod, L., Carpenter, D., Collins, L., Piwnica‐Worms, D., Hewitt, S., Udupi, G.M., Gallagher, W.M., Wegner, C., West, B.L., Wang‐Gillam, A., Goedegebuure, P., Linehan, D.C., and DeNardo, D.G. 2013. Targeting tumor‐infiltrating macrophages decreases tumor‐initiating cells, relieves immunosuppression, and improves chemotherapeutic responses. Cancer Res. 73:1128‐1141. doi: 10.1158/0008‐5472.CAN‐12‐2731.
  Motzer, R.J., Escudier, B., McDermott, D.F., George, S., Hammers, H.J., Srinivas, S., Tykodi, S.S., Sosman, J.A., Procopio, G., Plimack, E.R., Castellano, D., Choueiri, T.K., Gurney, H., Donskov, F., Bono, P., Wagstaff, J., Gauler, T.C., Ueda, T., Tomita, Y., Schutz, F.A., Kollmannsberger, C., Larkin, J., Ravaud, A., Simon, J.S., Xu, L.A., Waxman, I.M., Sharma, P., and CheckMate 025 Investigators. 2015. Nivolumab versus everolimus in advanced renal‐cell carcinoma. N. Engl. J. Med. 373:1803‐1813. doi: 10.1056/NEJMoa1510665.
  Murray, P.J., Allen, J.E., Biswas, S.K., Fisher, E.A., Gilroy, D.W., Goerdt, S., Gordon, S., Hamilton, J.A., Ivashkiv, L.B., Lawrence, T., Locati, M., Mantovani, A., Martinez, F.O., Mege, J.L., Mosser, D.M., Natoli, G., Saeij, J.P., Schultze, J.L., Shirey, K.A., Sica, A., Suttles, J., Udalova, I., van Ginderachter, J.A., Vogel, S.N., and Wynn, T.A. 2014. Macrophage activation and polarization: Nomenclature and experimental guidelines. Immunity 41:14‐20. doi: 10.1016/j.immuni.2014.06.008.
  Olive, K.P. and Tuveson, D.A. 2006. The use of targeted mouse models for preclinical testing of novel cancer therapeutics. Clin. Cancer Res. 12:5277‐5287. doi: 10.1158/1078‐0432.CCR‐06‐0436.
  Olive, K.P., Jacobetz, M.A., Davidson, C.J., Gopinathan, A., McIntyre, D., Honess, D., Madhu, B., Goldgraben, M.A., Caldwell, M.E., Allard, D., Frese, K.K., Denicola, G., Feig, C., Combs, C., Winter, S.P., Ireland‐Zecchini, H., Reichelt, S., Howat, W.J., Chang, A., Dhara, M., Wang, L., Ruckert, F., Grutzmann, R., Pilarsky, C., Izeradjene, K., Hingorani, S.R., Huang, P., Davies, S.E., Plunkett, W., Egorin, M., Hruban, R.H., Whitebread, N., McGovern, K., Adams, J., Iacobuzio‐Donahue, C., Griffiths, J., and Tuveson, D.A. 2009. Inhibition of Hedgehog signaling enhances delivery of chemotherapy in a mouse model of pancreatic cancer. Science 324:1457‐1461. doi: 10.1126/science.1171362.
  Perez‐Mancera, P.A., Guerra, C., Barbacid, M., and Tuveson, D.A. 2012. What we have learned about pancreatic cancer from mouse models. Gastroenterology 142:1079‐1092. doi: 10.1053/j.gastro.2012.03.002.
  Postow, M.A., Chesney, J., Pavlick, A.C., Robert, C., Grossmann, K., McDermott, D., Linette, G.P., Meyer, N., Giguere, J.K., Agarwala, S.S., Shaheen, M., Ernstoff, M.S., Minor, D., Salama, A.K., Taylor, M., Ott, P.A., Rollin, L.M., Horak, C., Gagnier, P., Wolchok, J.D., and Hodi, F.S. 2015. Nivolumab and ipilimumab versus ipilimumab in untreated melanoma. N. Engl. J. Med. 372:2006‐2017. doi: 10.1056/NEJMoa1414428.
  Provenzano, P.P., Cuevas, C., Chang, A.E., Goel, V.K., Von Hoff, D.D., and Hingorani, S.R. 2012. Enzymatic targeting of the stroma ablates physical barriers to treatment of pancreatic ductal adenocarcinoma. Cancer Cell 21:418‐429. doi: 10.1016/j.ccr.2012.01.007.
  Rei, M., Pennington, D.J., and Silva‐Santos, B. 2015. The emerging Protumor role of gammadelta T lymphocytes: Implications for cancer immunotherapy. Cancer Res. 75:798‐802. doi: 10.1158/0008‐5472.CAN‐14‐3228.
  Rhim, A.D., Mirek, E.T., Aiello, N.M., Maitra, A., Bailey, J.M., McAllister, F., Reichert, M., Beatty, G.L., Rustgi, A.K., Vonderheide, R.H., Leach, S.D., and Stanger, B.Z. 2012. EMT and dissemination precede pancreatic tumor formation. Cell 148:349‐361. doi: 10.1016/j.cell.2011.11.025.
  Rhim, A.D., Thege, F.I., Santana, S.M., Lannin, T.B., Saha, T.N., Tsai, S., Maggs, L.R., Kochman, M.L., Ginsberg, G.G., Lieb, J.G., Chandrasekhara, V., Drebin, J.A., Ahmad, N., Yang, Y.X., Kirby, B.J., and Stanger, B.Z. 2014. Detection of circulating pancreas epithelial cells in patients with pancreatic cystic lesions. Gastroenterology 146:647‐651. doi: 10.1053/j.gastro.2013.12.007.
  Robert, C., Long, G.V., Brady, B., Dutriaux, C., Maio, M., Mortier, L., Hassel, J.C., Rutkowski, P., McNeil, C., Kalinka‐Warzocha, E., Savage, K.J., Hernberg, M.M., Lebbe, C., Charles, J., Mihalcioiu, C., Chiarion‐Sileni, V., Mauch, C., Cognetti, F., Arance, A., Schmidt, H., Schadendorf, D., Gogas, H., Lundgren‐Eriksson, L., Horak, C., Sharkey, B., Waxman, I.M., Atkinson, V., and Ascierto, P.A. 2015a. Nivolumab in previously untreated melanoma without BRAF mutation. N. Engl. J. Med. 372:320‐330. doi: 10.1056/NEJMoa1412082.
  Robert, C., Schachter, J., Long, G.V., Arance, A., Grob, J.J., Mortier, L., Daud, A., Carlino, M.S., McNeil, C., Lotem, M., Larkin, J., Lorigan, P., Neyns, B., Blank, C.U., Hamid, O., Mateus, C., Shapira‐Frommer, R., Kosh, M., Zhou, H., Ibrahim, N., Ebbinghaus, S., and Ribas, A. 2015b. Pembrolizumab versus ipilimumab in advanced melanoma. N. Engl. J. Med 372:2521‐2532. doi: 10.1056/NEJMoa1503093.
  Rongvaux, A., Willinger, T., Martinek, J., Strowig, T., Gearty, S.V., Teichmann, L.L., Saito, Y., Marches, F., Halene, S., Palucka, A.K., Manz, M.G., and Flavell, R.A. 2014. Development and function of human innate immune cells in a humanized mouse model. Nat. Biotechnol. 32:364‐372. doi: 10.1038/nbt.2858.
  Rosenberg, S.A., Restifo, N.P., Yang, J.C., Morgan, R.A., and Dudley, M.E. 2008. Adoptive cell transfer: A clinical path to effective cancer immunotherapy. Nature Rev. Cancer 8:299‐308. doi: 10.1038/nrc2355.
  Royal, R.E., Levy, C., Turner, K., Mathur, A., Hughes, M., Kammula, U.S., Sherry, R.M., Topalian, S.L., Yang, J.C., Lowy, I., and Rosenberg, S.A. 2010. Phase 2 trial of single agent Ipilimumab (anti‐CTLA‐4) for locally advanced or metastatic pancreatic adenocarcinoma. J. Immunother. 33:828‐833. doi: 10.1097/CJI.0b013e3181eec14c.
  Roychoudhuri, R., Eil, R.L., and Restifo, N.P. 2015. The interplay of effector and regulatory T cells in cancer. Curr. Opin. Immunol. 33:101‐111. doi: 10.1016/j.coi.2015.02.003.
  Sastra, S.A. and Olive, K.P. 2013. Quantification of murine pancreatic tumors by high‐resolution ultrasound. Methods Mol. Biol. 980:249‐266. doi: 10.1007/978‐1‐62703‐287‐2_13.
  Schadendorf, D., Hodi, F.S., Robert, C., Weber, J.S., Margolin, K., Hamid, O., Patt, D., Chen, T.T., Berman, D.M., and Wolchok, J.D. 2015. Pooled analysis of long‐term survival data from phase II and Phase III trials of ipilimumab in unresectable or metastatic melanoma. J. Clin. Oncol. 33:1889‐1894. doi: 10.1200/JCO.2014.56.2736.
  Schreiber, R.D., Old, L.J., and Smyth, M.J. 2011. Cancer immunoediting: Integrating immunity's roles in cancer suppression and promotion. Science 331:1565‐1570. doi: 10.1126/science.1203486.
  Schumacher, T.N. and Schreiber, R.D. 2015. Neoantigens in cancer immunotherapy. Science 348:69‐74. doi: 10.1126/science.aaa4971.
  Siegel, R.L., Miller, K.D., and Jemal, A. 2015. Cancer statistics, 2015. CA Cancer J. Clin. 65:5‐29. doi: 10.3322/caac.21254.
  Singh, M., Lima, A., Molina, R., Hamilton, P., Clermont, A.C., Devasthali, V., Thompson, J.D., Cheng, J.H., BouReslan, H., Ho, C.C., Cao, T.C., Lee, C.V., Nannini, M.A., Fuh, G., Carano, R.A., Koeppen, H., Yu, R.X., Forrest, W.F., Plowman, G.D., and Johnson, L. 2010. Assessing therapeutic responses in Kras mutant cancers using genetically engineered mouse models. Nat. Biotechnol. 28:585‐593. doi: 10.1038/nbt.1640.
  Soares, K.C., Foley, K., Olino, K., Leubner, A., Mayo, S.C., Jain, A., Jaffee, E., Schulick, R.D., Yoshimura, K., Edil, B., and Zheng, L. 2014. A preclinical murine model of hepatic metastases. J. Vis. Exp. (91):51677. doi: 10.3791/51677.
  Sotomayor, E.M., Borrello, I., Tubb, E., Rattis, F.M., Bien, H., Lu, Z., Fein, S., Schoenberger, S., and Levitsky, H.I. 1999. Conversion of tumor‐specific CD4+ T‐cell tolerance to T‐cell priming through in vivo ligation of CD40. Nat. Med. 5:780‐787. doi: 10.1038/10503.
  Stromnes, I.M., Schmitt, T.M., Hulbert, A., Brockenbrough, J.S., Nguyen, H.N., Cuevas, C., Dotson, A.M., Tan, X., Hotes, J.L., Greenberg, P.D., and Hingorani, S.R. 2015. T cells engineered against a native antigen can surmount immunologic and physical barriers to treat pancreatic ductal adenocarcinoma. Cancer Cell 28:638‐652. doi: 10.1016/j.ccell.2015.09.022.
  Thomas, A.M., Santarsiero, L.M., Lutz, E.R., Armstrong, T.D., Chen, Y.C., Huang, L.Q., Laheru, D.A., Goggins, M., Hruban, R.H., and Jaffee, E.M. 2004. Mesothelin‐specific CD8(+) T cell responses provide evidence of in vivo cross‐priming by antigen‐presenting cells in vaccinated pancreatic cancer patients. J. Exp. Med. 200:297‐306. doi: 10.1084/jem.20031435.
  Topalian, S.L., Hodi, F.S., Brahmer, J.R., Gettinger, S.N., Smith, D.C., McDermott, D.F., Powderly, J.D., Carvajal, R.D., Sosman, J.A., Atkins, M.B., Leming, P.D., Spigel, D.R., Antonia, S.J., Horn, L., Drake, C.G., Pardoll, D.M., Chen, L., Sharfman, W.H., Anders, R.A., Taube, J.M., McMiller, T.L., Xu, H., Korman, A.J., Jure‐Kunkel, M., Agrawal, S., McDonald, D., Kollia, G.D., Gupta, A., Wigginton, J.M., and Sznol, M. 2012. Safety, activity, and immune correlates of anti‐PD‐1 antibody in cancer. N. Engl. J. Med. 366:2443‐2454. doi: 10.1056/NEJMoa1200690.
  Waddell, N., Pajic, M., Patch, A.M., Chang, D.K., Kassahn, K.S., Bailey, P., Johns, A.L., Miller, D., Nones, K., Quek, K., Quinn, M.C., Robertson, A.J., Fadlullah, M.Z., Bruxner, T.J., Christ, A.N., Harliwong, I., Idrisoglu, S., Manning, S., Nourse, C., Nourbakhsh, E., Wani, S., Wilson, P.J., Markham, E., Cloonan, N., Anderson, M.J., Fink, J.L., Holmes, O., Kazakoff, S.H., Leonard, C., Newell, F., Poudel, B., Song, S., Taylor, D., Waddell, N., Wood, S., Xu, Q., Wu, J., Pinese, M., Cowley, M.J., Lee, H.C., Jones, M.D., Nagrial, A.M., Humphris, J., Chantrill, L.A., Chin, V., Steinmann, A.M., Mawson, A., Humphrey, E.S., Colvin, E.K., Chou, A., Scarlett, C.J., Pinho, A.V., Giry‐Laterriere, M., Rooman, I., Samra, J.S., Kench, J.G., Pettitt, J.A., Merrett, N.D., Toon, C., Epari, K., Nguyen, N.Q., Barbour, A., Zeps, N., Jamieson, N.B., Graham, J.S., Niclou, S.P., Bjerkvig, R., Grutzmann, R., Aust, D., Hruban, R.H., Maitra, A., Iacobuzio‐Donahue, C.A., Wolfgang, C.L., Morgan, R.A., Lawlor, R.T., Corbo, V., Bassi, C., Falconi, M., Zamboni, G., Tortora, G., Tempero, M.A., Gill, A.J., Eshleman, J.R., Pilarsky, C., Scarpa, A., Musgrove, E.A., Pearson, J.V., Biankin, A.V., and Grimmond, S.M. 2015. Whole genomes redefine the mutational landscape of pancreatic cancer. Nature 518:495‐501. doi: 10.1038/nature14169.
  Westphalen, C.B. and Olive, K.P. 2012. Genetically engineered mouse models of pancreatic cancer. Cancer J. 18:502‐510. doi: 10.1097/PPO.0b013e31827ab4c4.
  Whatcott, C.J., Diep, C.H., Jiang, P., Watanabe, A., LoBello, J., Sima, C., Hostetter, G., Shepard, H.M., Von Hoff, D.D., and Han, H. 2015. Desmoplasia in primary tumors and metastatic lesions of pancreatic cancer. Clin. Cancer Res. 21:3561‐3568. doi: 10.1158/1078‐0432.CCR‐14‐1051.
  Winograd, R., Byrne, K.T., Evans, R.A., Odorizzi, P.M., Meyer, A.R., Bajor, D.L., Clendenin, C., Stanger, B.Z., Furth, E.E., Wherry, E.J., and Vonderheide, R.H. 2015. Induction of t‐cell immunity overcomes complete resistance to PD‐1 and CTLA‐4 blockade and improves survival in pancreatic carcinoma. Cancer Immunol. Res. 3:399‐411. doi: 10.1158/2326‐6066.CIR‐14‐0215.
  Zhang, L., Conejo‐Garcia, J.R., Katsaros, D., Gimotty, P.A., Massobrio, M., Regnani, G., Makrigiannakis, A., Gray, H., Schlienger, K., Liebman, M.N., Rubin, S.C., and Coukos, G. 2003. Intratumoral T cells, recurrence, and survival in epithelial ovarian cancer. N. Engl. J. Med. 348:203‐213. doi: 10.1056/NEJMoa020177.
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