1645P - Analysis of programmed death-ligand 1 (PD-L1) expression, transforming growth factor (TGF)-β gene expression signatures (GES) and tumor-infiltratin...

Date 11 September 2017
Event ESMO 2017 Congress
Session Poster display session
Topics Hepatobiliary Cancers
Translational Research
Presenter Yue Zhang
Citation Annals of Oncology (2017) 28 (suppl_5): v573-v594. 10.1093/annonc/mdx390
Authors Y. Zhang1, B.J. Naughton2, P..A. Rolfe3, E. Frick-Krieger4, I. Dussault5, L. Terracciano6, C. Ihling7
  • 1R&d Global Early Development, EMD Serono Research & Development Institute, 01821 - Billerica/US
  • 2Bioinformatics, EMD Serono Research & Development Institute, Billerica/US
  • 3R&d Global Early Development, EMD Serono Research & Development Institute, Billerica/US
  • 4Rd-ebp-h, Merck KGaA, Darmstadt/DE
  • 5Clinical Biomarkers And Companion Diagnostics, EMD Serono Research & Development Institute, Billerica/US
  • 6Pathology, Institute of Pathology, University Hospital Basel, Basel/CH
  • 7Cbd, Merck KGaA, Darmstadt/DE



HCC evades antitumor immune responses via multiple mechanisms, including the PD-L1 and TGF-β pathways. PD-L1 expression correlates with tumor aggressiveness and recurrence. Increased TGF-β activity corresponds with poor clinical outcomes. Using immunohistochemistry (IHC), we previously showed that PD-L1 expression in HCC stems primarily from IC. To further assess the HCC immune milieu, we measured IC, TGF-β-associated GES, and PD-L1 expression using IHC/RNAseq.


We assessed protein expression in 50 resected HCC specimens by quantitative (Q) IHC (primary antibodies: PD-L1, CD8, CD68) using standard techniques and automated software. For RNAseq, we prepared strand-specific libraries from extracted RNA, which were sequenced and compared to GES from published papers, CIBERSORT and Ingenuity Pathway Analysis.


All cases had typical morphology (low- to high-grade trabecular, pseudoglandular, or solid with common cytoplasmic features). Q CD8 IHC significantly correlated with CD8 mRNA expression and CD8 T cell GES, supporting the utility of RNAseq to evaluate the role of CD8+ T cells in HCC. RNAseq identified TGF-β1 as the main TGF-β isoform in HCC. Predefined TGF-β GES correlated strongly with EMT GES. There was a trend toward increased TGF-β1 activity and EMT marker expression in the S1 molecular subtype, which has previously been associated with TGF-β-driven aberrant Wnt signaling. Q CD8 IHC correlated with PD-L1 mRNA and protein levels in IC. In samples with high CD8, there was a trend of increased tumor-associated macrophages (TAMs); the presence of TAMs strongly correlated with TGF-β GES. Interestingly, few tumor cells displayed membranous PD-L1 staining as confirmed by PD-L1/pan-cytokeratin double labeling.


We used RNAseq and IHC to better understand the immunosuppressive environment in HCC driven by TGF-β and PD-L1, which may mediate different mechanisms to inhibit preexisting CD8+ IC.

Clinical trial identification


Legal entity responsible for the study

Funding was provided by Merck KGaA, Darmstadt, Germany.


Funding was provided by Merck KGaA, Darmstadt, Germany. Disclosure: Y. Zhang, B.J. Naughton, P.A. Rolfe, I. Dussault: Employee of EMD Serono Research & Development Institute, Billerica, MA, USA. E. Frick-Krieger, C. Ihling: Employee of Merck KGaA, Darmstadt, Germany. L. Terracciano: Consulting/advisory role to Merck AG