Antitumor effects of L-BLP25 Antigen-Specific tumor immunotherapy in a novel human MUC1 transgenic lung cancer mouse model

Journal of Translational Medicine

Michael W DeGregorio¹  Michael Wolf²  Chiao-Jung Kao¹  Stephen M Griffey³  Daniel P Vang¹  Brittany E Greenberg¹  Audrey M Gutierrez¹  Gregory T Wurz¹ 

[1] Department of Internal Medicine, Division of Hematology and Oncology, School of Medicine, University of California, Davis, 4501 X Street Suite 3016, Sacramento, CA, 95817, USA;ImmunoOncology, Merck Serono Research, Merck KGaA, Germany;Comparative Pathology Laboratory, UC Davis School of Veterinary Medicine, University of California, Davis, CA, USA

关键词: Cytokines;    Lung cancer;    Immunotherapy;    MUC1;    L-BLP25;   

Others  :  827957
DOI  :  10.1186/1479-5876-11-64

 received in 2012-12-01, accepted in 2013-03-03,  发布年份 2013

【 摘 要 】

Background

L-BLP25 antigen-specific cancer immunotherapeutic agent is currently in phase III clinical trials for non-small cell lung cancer. Using a novel human MUC1 transgenic (hMUC1.Tg) lung cancer mouse model, we evaluated effects of L-BLP25 combined with low-dose cyclophosphamide (CPA) pretreatment on Th1/Th2 cytokine production and antitumor activity.

Methods

A chemically-induced lung tumor model was developed in hMUC1.Tg C57BL/6 mice by administering 10 weekly 0.75-mg/g doses of the chemical carcinogen urethane by intraperitoneal injection. Serum cytokines associated with Th1/Th2 polarization and inflammation were measured by multiplex cytokine assay during tumorigenesis. Antitumor activity of L-BLP25 (10 μg) with CPA (100 mg/kg) pretreatment was evaluated following either one or two eight-week cycles of treatment by preparing lung whole mounts and counting tumor foci, and assessing IFN-γ production by ELISpot assay.

Results

During the carcinogenesis phase, no detectable Th1- or Th2-associated cytokine responses were observed, but levels of pro-inflammatory cytokines were increased with distinctive kinetics. A single cycle of L-BLP25 consisting of eight weekly doses was ineffective, whereas adding a second cycle given during tumor progression showed a significant reduction in the incidence of tumor foci. Administering two cycles of L-BLP25 induced Th1 cytokines IL-12, IL-2 and IFNγ at 24 h after the last dose, while Th2 and inflammatory cytokines were elevated to a lesser extent.

Conclusions

Urethane-induced lung tumors in hMUC1.Tg mice can be used as a model to assess the efficacy of the MUC1 antigen-specific cancer immunotherapeutic agent L-BLP25. The results indicate that the antitumor response to L-BLP25 requires at least two cycles and pre-treatment with CPA. In addition, monitoring pro-inflammatory serum cytokines may be useful as a biomarker of L-BLP25 response. Taken together, the preclinical lung tumor model can be utilized for determining effective combinations of L-BLP25 with chemotherapy and/or other immunotherapies.

【 授权许可】

   
2013 Wurz et al.; licensee BioMed Central Ltd.

【 图 表 】

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【 参考文献 】

• [1]Esposito L, Conti D, Ailavajhala R, Khalil N, Giordano A: Lung Cancer: Are we up to the Challenge? Curr Genomics 2010, 11:513-518.
• [2]Hirschowitz EA, Hiestand DM, Yannelli JR: Vaccines for lung cancer. J Thorac Oncol 2006, 1:93-104.
• [3]Nemunaitis J, Murray N: Immune-modulating vaccines in non-small cell lung cancer. J Thorac Oncol 2006, 1:756-761.
• [4]Barnd DL, Lan MS, Metzgar RS, Finn OJ: Specific, major histocompatibility complex-unrestricted recognition of tumor-associated mucins by human cytotoxic T cells. Proc Natl Acad Sci U S A 1989, 86:7159-7163.
• [5]Choi C, Witzens M, Bucur M, Feuerer M, Sommerfeldt N, Trojan A, Ho A, Schirrmacher V, Goldschmidt H, Beckhove P: Enrichment of functional CD8 memory T cells specific for MUC1 in bone marrow of patients with multiple myeloma. Blood 2005, 105:2132-2134.
• [6]Szabo E: MUC1 expression in lung cancer. Methods Mol Med 2003, 74:251-258.
• [7]Dong Y, Walsh MD, Cummings MC, Wright RG, Khoo SK, Parsons PG, McGuckin MA: Expression of MUC1 and MUC2 mucins in epithelial ovarian tumours. J Pathol 1997, 183:311-317.
• [8]Gao J, McConnell MJ, Yu B, Li J, Balko JM, Black EP, Johnson JO, Lloyd MC, Altiok S, Haura EB: MUC1 is a downstream target of STAT3 and regulates lung cancer cell survival and invasion. Int J Oncol 2009, 35:337-345.
• [9]Giatromanolaki A, Koukourakis MI, Sivridis E, O'Byrne K, Cox G, Thorpe PE, Gatter KC, Harris AL: Coexpression of MUC1 glycoprotein with multiple angiogenic factors in non-small cell lung cancer suggests coactivation of angiogenic and migration pathways. Clin Cancer Res 2000, 6:1917-1921.
• [10]Guddo F, Giatromanolaki A, Koukourakis MI, Reina C, Vignola AM, Chlouverakis G, Hilkens J, Gatter KC, Harris AL, Bonsignore G: MUC1 (episialin) expression in non-small cell lung cancer is independent of EGFR and c-erbB-2 expression and correlates with poor survival in node positive patients. J Clin Pathol 1998, 51:667-671.
• [11]Khodarev NN, Pitroda SP, Beckett MA, MacDermed DM, Huang L, Kufe DW, Weichselbaum RR: MUC1-induced transcriptional programs associated with tumorigenesis predict outcome in breast and lung cancer. Cancer Res 2009, 69:2833-2837.
• [12]Raina D, Kosugi M, Ahmad R, Panchamoorthy G, Rajabi H, Alam M, Shimamura T, Shapiro GI, Supko J, Kharbanda S, Kufe D: Dependence on the MUC1-C oncoprotein in non-small cell lung cancer cells. Mol Cancer Ther 2011, 10:806-816.
• [13]Mehta NR, Wurz GT, Burich RA, Greenberg BE, Griffey S, Gutierrez A, Bell KE, McCall JL, Wolf M, DeGregorio M: L-BLP25 vaccine plus letrozole induces a TH1 immune response and has additive antitumor activity in MUC1-expressing mammary tumors in mice. Clin Cancer Res 2012, 18:2861-2871.
• [14]Sangha R, Butts C: L-BLP25: a peptide vaccine strategy in non small cell lung cancer. Clin Cancer Res 2007, 13:s4652-s4654.
• [15]Salgaller ML: The development of immunotherapies for non-small cell lung cancer. Expert Opin Biol Ther 2002, 2:265-278.
• [16]Butts C, Maksymiuk A, Goss G, Soulieres D, Marshall E, Cormier Y, Ellis PM, Price A, Sawhney R, Beier F: Updated survival analysis in patients with stage IIIB or IV non-small-cell lung cancer receiving BLP25 liposome vaccine (L-BLP25): phase IIB randomized, multicenter, open-label trial. J Cancer Res Clin Oncol 2011, 137:1337-1342.
• [17]Ohyanagi F, Horai T, Sekine I, Yamamoto N, Nakagawa K, Nishio M, Senger S, Morsli N, Tamura T: Safety of BLP25 liposome vaccine (L-BLP25) in Japanese patients with unresectable stage III NSCLC after primary chemoradiotherapy: preliminary results from a Phase I/II study. Jpn J Clin Oncol 2011, 41:718-722.
• [18]Alberti W, Anderson G, Bartolucci A, Bell D, Villalba JB, Brodin O, Cardiello C, Cartei F, Cartei G, Cellerino R: Chemotherapy in Non-Small-Cell Lung-Cancer - a Metaanalysis Using Updated Data on Individual Patients from 52 Randomized Clinical-Trials. Br Med J 1995, 311:899-909.
• [19]Stearman RS, Dwyer-Nield L, Zerbe L, Blaine SA, Chan Z, Bunn PA Jr, Johnson GL, Hirsch FR, Merrick DT, Franklin WA: Analysis of orthologous gene expression between human pulmonary adenocarcinoma and a carcinogen-induced murine model. Am J Pathol 2005, 167:1763-1775.
• [20]Sweet-Cordero A, Mukherjee S, Subramanian A, You H, Roix JJ, Ladd-Acosta C, Mesirov J, Golub TR, Jacks T: An oncogenic KRAS2 expression signature identified by cross-species gene-expression analysis. Nat Genet 2005, 37:48-55.
• [21]Malkinson AM: Primary lung tumors in mice as an aid for understanding, preventing, and treating human adenocarcinoma of the lung. Lung Cancer 2001, 32:265-279.
• [22]Rowse GJ, Tempero RM, VanLith ML, Hollingsworth MA, Gendler SJ: Tolerance and immunity to MUC1 in a human MUC1 transgenic murine model. Cancer Res 1998, 58:315-321.
• [23]Tuveson DA, Jacks T: Modeling human lung cancer in mice: similarities and shortcomings. Oncogene 1999, 18:5318-5324.
• [24]Miller YE, Dwyer-Nield LD, Keith RL, Le M, Franklin WA, Malkinson AM: Induction of a high incidence of lung tumors in C57BL/6 mice with multiple ethyl carbamate injections. Cancer Lett 2003, 198:139-144.
• [25]Jiang YG, Chen JK, Wu ZL: Promotive effect of diethylstilbestrol on urethan-induced mouse lung tumorigenesis. Chemosphere 2000, 41:187-190.
• [26]Reagan-Shaw S, Nihal M, Ahmad N: Dose translation from animal to human studies revisited. FASEB J 2008, 22:659-661.
• [27]Nikitin AY, Alcaraz A, Anver MR, Bronson RT, Cardiff RD, Dixon D, Fraire AE, Gabrielson EW, Gunning WT, Haines DC: Classification of proliferative pulmonary lesions of the mouse: recommendations of the mouse models of human cancers consortium. Cancer Res 2004, 64:2307-2316.
• [28]Dwinell MB, Lugering N, Eckmann L, Kagnoff MF: Regulated production of interferon-inducible T-cell chemoattractants by human intestinal epithelial cells. Gastroenterology 2001, 120:49-59.
• [29]Barbon CM, Yang M, Wands GD, Ramesh R, Slusher BS, Hedley ML, Luby TM: Consecutive low doses of cyclophosphamide preferentially target Tregs and potentiate T cell responses induced by DNA PLG microparticle immunization. Cell Immunol 2010, 262:150-161.
• [30]Berd D, Mastrangelo MJ: Effect of low dose cyclophosphamide on the immune system of cancer patients: reduction of T-suppressor function without depletion of the CD8+ subset. Cancer Res 1987, 47:3317-3321.
• [31]Ercolini AM, Ladle BH, Manning EA, Pfannenstiel LW, Armstrong TD, Machiels JP, Bieler JG, Emens LA, Reilly RT, Jaffee EM: Recruitment of latent pools of high-avidity CD8(+) T cells to the antitumor immune response. J Exp Med 2005, 201:1591-1602.
• [32]Lutsiak ME, Semnani RT, De Pascalis R, Kashmiri SV, Schlom J, Sabzevari H: Inhibition of CD4(+)25+ T regulatory cell function implicated in enhanced immune response by low-dose cyclophosphamide. Blood 2005, 105:2862-2868.
• [33]Motoyoshi Y, Kaminoda K, Saitoh O, Hamasaki K, Nakao K, Ishii N, Nagayama Y, Eguchi K: Different mechanisms for anti-tumor effects of low- and high-dose cyclophosphamide. Oncol Rep 2006, 16:141-146.
• [34]Huyan XH, Lin YP, Gao T, Chen RY, Fan YM: Immunosuppressive effect of cyclophosphamide on white blood cells and lymphocyte subpopulations from peripheral blood of Balb/c mice. Int Immunopharmacol 2011, 11:1293-1297.
• [35]Malvicini M, Alaniz L, Bayo J, Garcia M, Piccioni F, Fiore E, Atorrasagasti C, Aquino JB, Matar P, Mazzolini G: Single low-dose cyclophosphamide combined with interleukin-12 gene therapy is superior to a metronomic schedule in inducing immunity against colorectal carcinoma in mice. OncoImmunology 2012, 1:1-10.
• [36]Rabinovich GA, Rubinstein N, Matar P, Rozados V, Gervasoni S, Scharovsky GO: The antimetastatic effect of a single low dose of cyclophosphamide involves modulation of galectin-1 and Bcl-2 expression. Cancer Immunol Immunother 2002, 50:597-603.
• [37]Mukherjee P, Ginardi AR, Madsen CS, Tinder TL, Jacobs F, Parker J, Agrawal B, Longenecker BM, Gendler SJ: MUC1-specific CTLs are non-functional within a pancreatic tumor microenvironment. Glycoconj J 2001, 18:931-942.
• [38]Turner MS, Cohen PA, Finn OJ: Lack of effective MUC1 tumor antigen-specific immunity in MUC1-transgenic mice results from a Th/T regulatory cell imbalance that can be corrected by adoptive transfer of wild-type Th cells. J Immunol 2007, 178:2787-2793.
• [39]Machiels JP, Reilly RT, Emens LA, Ercolini AM, Lei RY, Weintraub D, Okoye FI, Jaffee EM: Cyclophosphamide, doxorubicin, and paclitaxel enhance the antitumor immune response of granulocyte/macrophage-colony stimulating factor-secreting whole-cell vaccines in HER-2/neu tolerized mice. Cancer Res 2001, 61:3689-3697.
• [40]Schlom J: Recent advances in therapeutic cancer vaccines. Cancer Biother Radiopharm 2012, 27:2-5.
• [41]DeGregorio M, Wurz GT, Gutierrez A, Wolf M: L-BLP25 vaccine plus letrozole for breast cancer: Is translation possible? OncoImmunology 2012, 1:1422-1424.