Expression of Checkpoint Molecules in the Tumor Microenvironment of Intrahepatic Cholangiocarcinoma: Implications for Immune Checkpoint Blockade Therapy
Lara Heij,
Jan Bednarsch,
Xiuxiang Tan,
Mika Rosin,
Simone Appinger,
Konrad Reichel,
Dana Pecina,
Michail Doukas,
Ronald M. van Dam,
Juan Garcia Vallejo,
Florian Ulmer,
Sven Lang,
Tom Luedde,
Flavio G. Rocha,
Shivan Sivakumar,
Ulf Peter Neumann
Affiliations
Lara Heij
Department of Surgery and Transplantation, University Hospital RWTH Aachen, 52074 Aachen, Germany
Jan Bednarsch
Department of Surgery and Transplantation, University Hospital RWTH Aachen, 52074 Aachen, Germany
Xiuxiang Tan
Department of Surgery and Transplantation, University Hospital RWTH Aachen, 52074 Aachen, Germany
Mika Rosin
Department of Surgery and Transplantation, University Hospital RWTH Aachen, 52074 Aachen, Germany
Simone Appinger
Department of Surgery and Transplantation, University Hospital RWTH Aachen, 52074 Aachen, Germany
Konrad Reichel
Department of Surgery and Transplantation, University Hospital RWTH Aachen, 52074 Aachen, Germany
Dana Pecina
Department of Surgery and Transplantation, University Hospital RWTH Aachen, 52074 Aachen, Germany
Michail Doukas
Department of Pathology, Erasmus Medical Center Rotterdam, 3000 CB Rotterdam, The Netherlands
Ronald M. van Dam
Department of Surgery and Transplantation, University Hospital RWTH Aachen, 52074 Aachen, Germany
Juan Garcia Vallejo
Department of Molecular Cell Biology & Immunology, VU University Medical Center, 1081 HZ Amsterdam, The Netherlands
Florian Ulmer
Department of Surgery and Transplantation, University Hospital RWTH Aachen, 52074 Aachen, Germany
Sven Lang
Department of Surgery and Transplantation, University Hospital RWTH Aachen, 52074 Aachen, Germany
Tom Luedde
Department of Gastroenterology, Hepatology and Infectious Diseases, University Hospital Duesseldorf, 40225 Duesseldorf, Germany
Flavio G. Rocha
Division of Surgical Oncology, Knight Cancer Institute, Oregon Health and Science University, Portland, OR 97239, USA
Shivan Sivakumar
Department of Oncology, University of Oxford, Oxford OX3 7DQ, UK
Ulf Peter Neumann
Department of Surgery and Transplantation, University Hospital RWTH Aachen, 52074 Aachen, Germany
Background: The tumor microenvironment (TME) in cholangiocarcinoma (CCA) influences the immune environment. Checkpoint blockade is promising, but reliable biomarkers to predict response to treatment are still lacking. Materials and Methods: The levels of checkpoint molecules (PD-1, PD-L1, PD-L2, LAG-3, ICOS, TIGIT, TIM-3, CTLA-4), macrophages (CD68), and T cells (CD4 and CD8 cells) were assessed by multiplexed immunofluorescence in 50 intrahepatic cases. Associations between marker expression, immune cells, and region of expression were studied in the annotated regions of tumor, interface, sclerotic tumor, and tumor-free tissue. Results: ICCA demonstrated CD4_TIM-3 high densities in the tumor region of interest (ROI) compared to the interface (p = 0.014). CD8_PD-L1 and CD8_ICOS densities were elevated in the sclerotic tumor compared to the interface (p = 0.011 and p = 0.031, respectively). In a multivariate model, high expression of CD8_PD-L2 (p = 0.048) and CD4_ICOS_TIGIT (p = 0.011) was associated with nodal metastases. Conclusions: High densities of PD-L1 were more abundant in the sclerotic tumor region; this is meaningful for the stratification of immunotherapy. Lymph node metastasis correlates with CD4_ICOS_TIGIT co-expression and CD8_PD-L2 expression, indicating the checkpoint expression profile of patients with a poor prognosis. Also, multiple co-expressions occur, and this potentially suggests a role for combination therapy with different immune checkpoint targets than just PD-1 blockade monotherapy.
