PMID- 33424844 OWN - NLM STAT- MEDLINE DCOM- 20210628 LR - 20210628 IS - 1664-3224 (Electronic) IS - 1664-3224 (Linking) VI - 11 DP - 2020 TI - PD-L1 and PD-L2 Expression in Cervical Cancer: Regulation and Biomarker Potential. PG - 596825 LID - 10.3389/fimmu.2020.596825 [doi] LID - 596825 AB - PD-1/PD-L1 immune checkpoint inhibitors show potential for cervical cancer treatment. However, low response rates suggest that patient selection based on PD-L1 protein expression is not optimal. Here, we evaluated different PD-L1 detection methods and studied transcriptional regulation of PD-L1/PD-L2 expression by The Cancer Genome Atlas (TCGA) mRNAseq analysis. First, we determined the copy number of the PD-L1/PD-L2 locus by fluorescence in situ hybridization (FISH), PD-L1 mRNA expression by RNA in situ hybridization (RNAish), and PD-L1/PD-L2 protein expression by immunohistochemistry (IHC) on tissue microarrays containing a cohort of 60 patients. Additionally, distribution of PD-L1/PD-L2 was visualized based on flow cytometry analysis of single-cell suspensions (n = 10). PD-L1/PD-L2 locus amplification was rare (2%). PD-L1 mRNA expression in tumor cells was detected in 56% of cases, while 41% expressed PD-L1 protein. Discordant scores for PD-L1 protein expression on tumor cells between cores from one patient were observed in 27% of cases. Interestingly, with RNAish, PD-L1 heterogeneity was observed in only 11% of the cases. PD-L2 protein expression was found in 53%. PD-L1 mRNA and protein expression on tumor cells were strongly correlated (p < 0.001). PD-L1 and PD-L2 protein expression showed no correlation on tumor cells (p = 0.837), but a strong correlation on cells in stromal fields (p < 0.001). Co-expression of PD-L1 and PD-L2 on macrophage-like populations was also observed with flow cytometry analysis. Both PD-L1 and PD-L2 TCGA transcript levels strongly correlated in the TCGA data, and both PD-L1 and PD-L2 strongly correlated with interferon gamma (IFNG) expression/transcript levels (p < 0.0001). Importantly, patients with high PD-L1/PD-L2/IFNG transcript levels had a survival advantage over patients with high PD-L1/PD-L2 and low IFNG expression. Based on these findings, we conclude that PD-L1/PD-L2 expression in cervical cancer is mainly associated with interferon induction and not gene amplification, which makes FISH unsuitable as biomarker. The heterogeneous PD-L1 and PD-L2 expression patterns suggest IHC unreliable for patient selection. RNAish, in conjunction with interferon signaling evaluation, seems a promising technique for immune checkpoint detection. These results warrant further investigation into their prognostic and predictive potential. CI - Copyright (c) 2020 Rotman, Otter, Bleeker, Samuels, Heeren, Roemer, Kenter, Zijlmans, van Trommel, de Gruijl and Jordanova. FAU - Rotman, Jossie AU - Rotman J AD - Center for Gynecologic Oncology Amsterdam (CGOA), Amsterdam University Medical Center (UMC), Cancer Center Amsterdam (CCA), Vrije Universiteit Amsterdam, Amsterdam, Netherlands. AD - Department of Medical Oncology Amsterdam UMC, Cancer Center Amsterdam (CCA), Vrije Universiteit Amsterdam, Amsterdam, Netherlands. FAU - den Otter, Leontine A S AU - den Otter LAS AD - Center for Gynecologic Oncology Amsterdam (CGOA), Amsterdam University Medical Center (UMC), Cancer Center Amsterdam (CCA), Vrije Universiteit Amsterdam, Amsterdam, Netherlands. FAU - Bleeker, Maaike C G AU - Bleeker MCG AD - Department of Pathology, Cancer Center Amsterdam (CCA), Amsterdam UMC, Vrije Universiteit Amsterdam, Amsterdam, Netherlands. FAU - Samuels, Sanne S AU - Samuels SS AD - Center for Gynecologic Oncology Amsterdam (CGOA), Amsterdam University Medical Center (UMC), Cancer Center Amsterdam (CCA), Vrije Universiteit Amsterdam, Amsterdam, Netherlands. FAU - Heeren, A Marijne AU - Heeren AM AD - Department of Medical Oncology Amsterdam UMC, Cancer Center Amsterdam (CCA), Vrije Universiteit Amsterdam, Amsterdam, Netherlands. FAU - Roemer, Margaretha G M AU - Roemer MGM AD - Department of Pathology, Cancer Center Amsterdam (CCA), Amsterdam UMC, Vrije Universiteit Amsterdam, Amsterdam, Netherlands. FAU - Kenter, Gemma G AU - Kenter GG AD - Center for Gynecologic Oncology Amsterdam (CGOA), Amsterdam University Medical Center (UMC), Cancer Center Amsterdam (CCA), Vrije Universiteit Amsterdam, Amsterdam, Netherlands. AD - Center for Gynecologic Oncology Amsterdam (CGOA), Netherlands Cancer Institute-Antoni van Leeuwenhoek (NKI-AVL), Amsterdam, Netherlands. FAU - Zijlmans, Henry J M A A AU - Zijlmans HJMAA AD - Center for Gynecologic Oncology Amsterdam (CGOA), Netherlands Cancer Institute-Antoni van Leeuwenhoek (NKI-AVL), Amsterdam, Netherlands. FAU - van Trommel, Nienke E AU - van Trommel NE AD - Center for Gynecologic Oncology Amsterdam (CGOA), Netherlands Cancer Institute-Antoni van Leeuwenhoek (NKI-AVL), Amsterdam, Netherlands. FAU - de Gruijl, Tanja D AU - de Gruijl TD AD - Department of Medical Oncology Amsterdam UMC, Cancer Center Amsterdam (CCA), Vrije Universiteit Amsterdam, Amsterdam, Netherlands. FAU - Jordanova, Ekaterina S AU - Jordanova ES AD - Center for Gynecologic Oncology Amsterdam (CGOA), Amsterdam University Medical Center (UMC), Cancer Center Amsterdam (CCA), Vrije Universiteit Amsterdam, Amsterdam, Netherlands. AD - Department of Pathology, Leiden University Medical Center, Leiden, Netherlands. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20201217 PL - Switzerland TA - Front Immunol JT - Frontiers in immunology JID - 101560960 RN - 0 (B7-H1 Antigen) RN - 0 (Biomarkers, Tumor) RN - 0 (CD274 protein, human) RN - 0 (PDCD1LG2 protein, human) RN - 0 (Programmed Cell Death 1 Ligand 2 Protein) SB - IM MH - Adult MH - B7-H1 Antigen/*genetics/metabolism MH - *Biomarkers, Tumor MH - DNA Copy Number Variations MH - Female MH - *Gene Expression MH - Genetic Loci MH - High-Throughput Nucleotide Sequencing MH - Humans MH - Immunohistochemistry MH - Immunophenotyping MH - In Situ Hybridization, Fluorescence MH - Middle Aged MH - Programmed Cell Death 1 Ligand 2 Protein/*genetics/metabolism MH - Uterine Cervical Neoplasms/diagnosis/*genetics/metabolism PMC - PMC7793653 OTO - NOTNLM OT - RNAish OT - The Cancer Genome Atlas OT - cervical cancer OT - fluorescence in situ hybridization OT - immunohistochemistry OT - programmed cell death ligand 1 OT - programmed cell death ligand 2 COIS- The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. EDAT- 2021/01/12 06:00 MHDA- 2021/06/29 06:00 PMCR- 2020/01/01 CRDT- 2021/01/11 05:37 PHST- 2020/08/20 00:00 [received] PHST- 2020/11/16 00:00 [accepted] PHST- 2021/01/11 05:37 [entrez] PHST- 2021/01/12 06:00 [pubmed] PHST- 2021/06/29 06:00 [medline] PHST- 2020/01/01 00:00 [pmc-release] AID - 10.3389/fimmu.2020.596825 [doi] PST - epublish SO - Front Immunol. 2020 Dec 17;11:596825. doi: 10.3389/fimmu.2020.596825. eCollection 2020.