PD-L1 expression in cerebrospinal fluid for leptomeningeal metastasis from solid tumors: preliminary assessment of clinical implications

Zhenyu PanHang LeiYushan HuangMin LiuMiaomiao LiuPanpan TaiXiao ChenYu WuLishi WangYuancheng ZhangGuozi Yang^*

• The Affiliated Huizhou Hospital, Guangzhou Medical University, Huizhou, China

Background: Intrathecal immune checkpoint inhibitors have shown promise for leptomeningeal metastasis (LM). Programmed cell death ligand 1 (PD-L1) is currently the primary predictive biomarker for immunotherapy response. This study assessed the feasibility of PD-L1 detection via immunocytochemistry based on ThinPrep liquid-based cytology (LBC), explored its application in cerebrospinal fluid (CSF) from LM patients and preliminarily evaluated clinical implications. Methods: Technical validation used six human tumor cell lines (lung, breast and gastric) with validated high/low PD-L1 expression, which were processed into ThinPrep LBC slides and cell block sections. PD-L1 immunocytochemistry and immunohistochemistry were performed using four antibodies (Dako 22C3, Ventana SP263, Abcam 28-8 and SP142). Clinically, CSF samples from LM patients were ThinPrep-processed and PD-L1 immunocytochemistry-stained. PD-L1 expression was assessed via tumor proportion score. Concordance of different antibody assays, as well as that of PD-L1 expression between paired CSF and extracranial lesions, was analyzed using Cohen’s κ. The association between CSF PD-L1 expression and response to intrathecal immunotherapy was assessed. Results: Immunocytochemistry based on ThinPrep LBC reliably detected membrane PD-L1, showing high concordance with immunohistochemistry on cell blocks for clones 22C3/SP263 across all cell lines (NCI-H358, MDA-MB-231, SGC-7901, A549, MCF7, and AGS). Clones 28-8/SP142 showed false positives and were excluded. Using clones 22C3 /SP263, 130 CSF samples from 65 LM patients (55 lung, 8 breast, and 2 gastric cancers) were analyzed. Overall PD-L1 positivity rates were 48% with 22C3 and 51% with SP263 (κ=0.815). In the subset of 68 slides containing ≥100 tumor cells, PD-L1 positivity rates were 53% with 22C3 and 59% with SP263 (κ=0.881). For 62 slides containing 20-100 tumor cells, the rates were 42% with both antibodies (κ=0.735). PD-L1 expression showed poor agreement between 29 paired CSF and extracranial lesions (κ=0.175 with 22C3; κ=0.179 with SP263). Among 45 patients receiving intrathecal immunotherapy, A numerical increase in response rate was observed in PD-L1-positive patients (61.9% vs. 33.3%; p=0.055). Conclusions: Our study establishes a robust methodology for detecting CSF PD-L1 expression using ThinPrep LBC with immunocytochemistry for LM patients. This approach suggests potential utility of CSF PD-L1 expression as a biomarker for guiding intrathecal immunotherapy for LM from solid tumors.