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C06: Targeting the mutanome of liver cancer by viral oncolysis and tumor-directed immunotherapies

Viroimmunotherapy is an innovative therapy that has shown first successful therapeutic results in advanced melanoma but failed to improve overall survival in a phase IIb study in HCC. We have shown in previous work that oncolytic viruses induce inflammation of the tumor tissue and cross-presentation of tumor antigens providing excellent preconditions for immunotherapies such as immune checkpoint blockade. To investigate neoepitope-specific CD8 T-cell responses upon virotherapy and pharmacologic inhibition of the PD-1/PD-L1 axis, we have determined the neoepitope spectrum by transcriptomic sequencing and epitope prediction in CMT64 lung cancer cells. In s.c. implanted syngeneic tumors we showed that localized virotherapy with simultaneous blockade of the PD-1/PD-L1 axis using a PD-1-antagonistic antibody, led to a strong epitope-spreading of CD8 T-cell responses associated with improved immune-mediated elimination of lung metastases demonstrating that viral oncolysis overcomes systemic tumor resistance to PD-1/PD-L1 immunotherapy. Importantly, therapeutic efficacy of virotherapy and PD-1/PD-L1 checkpoint blockade was confirmed in a transgenic liver tumor model with spontaneous lung metastasis. In this project, we will investigate epitope spreading in response to oncolytic virotherapy and PD-1/PD-L1 checkpoint inhibition in mouse models with subcutaneously grown tumors of established murine HCC cell lines or self-generated HCC cell lines expressing a predefined CD4 and CD8 T-cell neoepitope spectrum. Furthermore, we will investigate the patterns of neoepitope-specific T-cell responses triggered by different oncolytic viruses (adenovirus, measles, vaccinia virus) and the influence of different immunostimulatory transgenes in epitope spreading. These investigations are urgently required to develop suitable immunotherapeutic strategies using virotherapy and immune checkpoint inhibition for liver cancers.


Knocke S, Fleischmann-Mundt B, Saborowski M, Manns MP, Kühnel F, Wirth TC, Woller N (2016) Tailored tumor immunogenicity reveals regulation of CD4 and CD8 T-cell responses against cancer. Cell Rep, 22;17(9):2234-2246 .
Gürlevik E, Fleischmann-Mundt B, Brooks J, Demir IE, Steiger K, Ribback S, Yevsa T, Woller N, Kloos A, Ostroumov D, Armbrecht N, Manns MP, Dombrowski F, Saborowski M, Kleine M, Wirth TC, Oettle H, Ceyhan GO, Esposito I, Calvisi DF, Kubicka S, Kühnel F (2016) NK cells prevent local recurrence after postoperative chemotherapy of pancreatic cancer with Akt2-driven metastasis. Gastroenterology 151(2):338-350.
Blake SJ, Stannard K, Liu J, Allen S, Yong MC, Mittal D, Roman Aguilera A, Miles JJ, Lutzky VP, Ferrari de Andrade L, Martinet L, Colonna M, Takeda K, Kühnel F, Gurlevik E, Bernhardt G, Teng MW, Smyth MJ (2016) Suppression of metastases using a new lymphocyte checkpoint target for cancer immunotherapy. Cancer Discov 6(4):446-59.
Woller N, Gürlevik E, Fleischmann-Mundt B, Knocke S, Kloos A, Saborowski M, Geffers R, Manns MP, Wirth TC, Kubicka S, Kühnel F (2015) Virotherapy overcomes tumor resistance to PD-1- immunotherapy by broad mutanome-directed T-cell responses in mice. Mol Ther 23(10):1630-40.
Kloos A, Woller N, Gürlevik E, Ureche CI, Armbrecht N, Geffers R, Manns MP, Gerardy-Schahn R, Kühnel F (2015) PolySia-specific retargeting of oncolytic viruses triggers tumor-specific immune responses and facilitates therapy of small cell lung cancer. Cancer Immunol Res (7):751-63.
Anggakusuma NFN, Frentzen A, Gürlevik E, Yuan Q, Steinmann E, Ott M, Staeheli P, Hornung V, Kühnel F, Pietschmann T (2015) Control of HCV replication in mouse liver derived cells by MAVSdependent production of type-I and -III interferons. J Virol 89(7):3833-45.
Gürlevik E, Fleischmann-Mundt B, Armbrecht N, Longerich T, Woller N, Kloos A, Hoffmann D, Schambach A, Manns MP, Zender L, Kubicka S*, Kühnel F* (2013) Adjuvant gemcitabine therapy improves survival in a locally induced, R0-resectable model of metastatic intrahepatic cholangiocarcinoma. Hepatology 58(3):1031-41.
Gürlevik E, Schache P, Goez A, Kloos A, Woller N, Armbrecht N, Manns MP, Kubicka S*, Kühnel F* (2013) Meganuclease-mediated virus self-cleavage facilitates tumor-specific replication. Mol There 21(9):1738-48.
Woller N, Knocke S, Mundt B, Gurlevik E, Struver N, Kloos A, Boozari B, Schache P, Manns MP, Malek NP, Sparwasser T, Zender L, Wirth TC, Kubicka S*, Kühnel F* (2011) Virus-induced tumor inflammation facilitates effective DC cancer immunotherapy in a Treg-dependent manner in mice. J Clin Invest 121(7): 2570-82.
Boozari B, Mundt B, Woller N, Strüver N, Gürlevik E, Schache P, Kloos A, Knocke S, Manns MP, Wirth TC, Kubicka S*, Kühnel F* (2010) Proteasome inhibition during oncolytic infection improves antitumoral immunity and inhibits metastatic growth of HCC. GUT 59:1416-1426. * Equal contribution of authors