25Transcriptome profiling identifies TIGIT and PD-1 co-expression in the immunosuppressive environment in cholangiocarcinoma

The Oncologist, vol. 31

Abstract

Abstract Background Checkpoint blockade in cholangiocarcinoma (CCA) is promising; however, little is known about the poor response. We investigated the single-cell and spatial immune environment in combination with checkpoint expression in CCA. Methods Checkpoint molecules (PD-1, PD-L 1, PD-l2, LAG-3, ICOS, TIGIT, TIM-3, and CTLA-4), macrophages (CD68), and T cells (CD4 and CDS) were assessed by multiplex immunofluorescence (mlF) (n = 50). We investigated the transcriptomic profile using the NanoString Cancer Progression Panel on selected TIGIT high and low samples (n = 12). Validation was performed by mlF (n = 24). Furthermore, we identified an immune profile with co-expression ofTIGIT and PD-1 in our Cytometry-Time-Of-Flight (CyTOF) cohort in 50% (n = 16). By snRNA sequencing (n = 6), we investigated cell-cell interactions and crosstalk mechanisms. The mouse model demonstrated representative immune cell populations, although dual blockade therapy (anti-TIGIT and PD-1) did not reach significance. Results The expression of checkpoint molecules TIGIT, CTLA-4, and LAG-3 alone and in combination with other checkpoint molecules was more abundant in the tumor than in normal tissue (CD4 and CDS TIGIT p < 0.0001, CD4 and CDS CTLA-4, p < 0.0001 and p < 0.001, respectively, and CDS LAG-3 p < 0.05). MMP2 and MMP14 genes were differentially expressed in patients with high TIGIT expression. PD-1 and TIGIT dual expression on CD4 and CDS cells are common in CCA. Conclusion The immune environment in CCA is characterized by the expression of multiple checkpoints, demonstrating the complexity of ICI treatment. High TIGIT expression drives an immunosuppressive environment by modulating the extracellular matrix. These findings suggest that CCA is unlikely to respond optimally to PD-1 monotherapy and support TIGIT as a promising target for combination immunotherapeutic strategies. Our cohort identified 50% of patients with high levels of dual checkpoint expression, stratifying a subgroup likely to respond to dual blockade therapy.

Authors 11

1. Lara Heij

   Essen University Hospital

   Affiliation as printed

   Department of General, Visceral and Transplant Surgery, University Hospital Essen , 45147 Essen,

   Department of Pathology, University Hospital Essen ,

2. Sikander Hayat Aachen Department of Renal and Hypertensive Disorders Rheumatological and Immunological Diseases (Medical Clinic II) Medical Faculty

   RWTH Aachen University

   Affiliation as printed

   Department of Renal and Hypertensive Disorders, Rheumatological and Immunological Diseases (Medical Clinic II), Medical Faculty, RWTH Aachen University , Aachen,

3. Konrad Reichel

   Essen University Hospital · Amsterdam UMC Location Vrije Universiteit Amsterdam

   Affiliation as printed

   Department of General, Visceral and Transplant Surgery, University Hospital Essen , 45147 Essen,

   Department of Molecular Cell Biology & Immunology, VU University Medical Center , Amsterdam,

4. Sidrah Maryam Aachen Department of Renal and Hypertensive Disorders Rheumatological and Immunological Diseases (Medical Clinic II) Medical Faculty

   RWTH Aachen University

   Affiliation as printed

   Department of Renal and Hypertensive Disorders, Rheumatological and Immunological Diseases (Medical Clinic II), Medical Faculty, RWTH Aachen University , Aachen,

5. Juan J. Garcia Vallejo

   Amsterdam UMC Location Vrije Universiteit Amsterdam

   Affiliation as printed

   Department of Molecular Cell Biology & Immunology, VU University Medical Center , Amsterdam,

6. Danny David Jonigk Aachen Institute of Pathology

   Universitätsklinikum Aachen · RWTH Aachen University

   Affiliation as printed

   Institute of Pathology, University Hospital RWTH Aachen , Aachen,

7. Mark Kuehnel Aachen Institute of Pathology

   Universitätsklinikum Aachen · RWTH Aachen University

   Affiliation as printed

   Institute of Pathology, University Hospital RWTH Aachen , Aachen,

8. Lueong Smiths

   Essen University Hospital

   Affiliation as printed

   Bridge Institute for Experimental Cancer Therapy, West German Cancer Center, University Hospital Essen , Hufeland Str. 55 , 45147, Essen,

9. J Hammons

   Emory University · Winship Cancer Institute

   Affiliation as printed

   Department of Hematology and Medical Oncology, Winship Cancer Institute of Emory University , Atlanta, GA,

10. Gregory B. Lesinski

    Emory University · Winship Cancer Institute

    Affiliation as printed

    Department of Hematology and Medical Oncology, Winship Cancer Institute of Emory University , Atlanta, GA,

11. Ulf Neumann

    Essen University Hospital

    Affiliation as printed

    Department of General, Visceral and Transplant Surgery, University Hospital Essen , 45147 Essen,