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C05: Microsphere-based immunotherapy of HCC

Hepatocellular carcinoma is increasingly recognized as an immunogenic tumor that represents a promising target for immunotherapy. In recent clinical trials, checkpoint blockade has induced durable remissions in a subset of patients treated with anti-PD-1 antibodies and currently the therapeutic value of both PD-1 and PD-L1 targeted immunotherapy is evaluated in clinical trials. The success of targeted vaccines, however, has so far been disappointing due to the immunosuppressive liver micromilieu which prevents induction of efficacious adaptive immune responses. We have established a prime-boost vaccination based on a primary immunization with antigen-coated polylactic co-glycolic acid (PLGA) microspheres and a secondary infection with Listeria monocytogenes (LM), a vector that does not induce humoral immunity in humans. Compared to conventional vaccines consisting of homologous immunizations of a combination of the target antigen with adjuvants, PLGA-LM vaccinations are by far more potent and induce cytolytic CD8 T cell immune responses in less than 14 days. The aim of the research proposal is to develop novel, heterologous PLGA/LM prime-boost vaccinations for the treatment of hepatocellular carcinoma and to test our therapeutic vaccination in the context of an orthotopic liver cancer model. To this extent, we will target glypican-3, a GPI-anchored surface protein that is frequently overexpressed in HCC and incorporate the target antigen into Listeria monocytogenes vectors. Using the orthotopic murine liver cancer model, we will furthermore aim at identifying suppressors of CD8 T cell function in the context of a Listeria-based vaccine. For this purpose we have performed Listeria vaccinations in mice bearing advanced, diffuse HCC and isolated exhausted HCC-specific CD8 T cells from the tumor-bearing mice. From these T cells we were able to assess the first whole genome transcriptomic signature of HCCspecific exhausted T cells and comparative mRNA expression profiles from naive and functional effectors CD8 T cells. For the research project proposed, we will make use of these unique microarray profiles to identify novel regulators of T cell exhaustion that could be exploited for therapeutic purposes. Furthermore, we will screen for surface markers on HCC-specific T cells that allow for the identification of tumorspecific T cells in peripheral blood samples without a priori knowledge of their cognate antigen.


Nimanong S, Ostroumov D, Wingerath J, Knocke S, Woller N, Gürlevik E, Falk CS, Manns MP, Kühnel F, Wirth TC. (2017) CD40 Signaling Drives Potent Cellular Immune Responses in Heterologous Cancer Vaccinations. Cancer Res. 77(8):1918-1926.
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.
Wirth TC, Manns MP (2016) The impact of the revolution in hepatitis C treatment on hepatocellular carcinoma. Ann Oncol 27(8):1467-74.
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) Administration of Gemcitabine After Pancreatic Tumor Resection in Mice Induces an Antitumor Immune Response Mediated by Natural Killer Cells. Gastroenterology 151(2):338-350.
Woller N, Gürlevik E, Fleischmann-Mundt B, Schumacher A, Knocke S, Kloos AM, Saborowski M, Geffers R, Manns MP, Wirth TC, Kubicka S, Kühnel F (2015) Viral Infection of Tumors Overcomes Resistance to PD-1-immunotherapy by Broadening Neoantigenome-directed T-cell Responses. Mol Ther 23(10):1630-40.
Brinkhoff B, Wirth TC (2014) Of microspheres and microbes: A double-hit strategy for cancer immunotherapy. Oncoimmunology 14;3:e27873.
Wirth TC (2014) Spontaneous and therapeutic immune responses in hepatocellular carcinoma: implications for current and future immunotherapies. Expert Rev Gastroenterol Hepatol 8(1):101-10.
Brinkhoff B, Ostroumov D, Heemcke J, Woller N, Gürlevik E, Manns MP, Longerich T, Zender L, Harty JT, Kubicka S, Kühnel F, Wirth TC (2014) Microsphere priming facilitates induction of potent therapeutic T cell immune responses against autochthonous liver cancers. Eur J Immunol 44(4):1213-24.
Gürlevik E, Fleischmann-Mundt B, Armbrecht N, Longerich T, Woller N, Kloos A, Hoffmann D, Schambach A, Wirth TC, 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.
Woller N, Knocke S, Mundt B, Gürlevik E, Strüver 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.
Wirth TC, Xue HH, Rai D, Sabel JT, Bair T, Harty JT, Badovinac VP (2010) Repetitive antigen stimulation induces stepwise transcriptome diversification but preserves a core signature of memory CD8(+) T cell differentiation. Immunity 33(1):128-40.