MiR-200a regulates PD-L1 and predicts response to immune checkpoint inhibitors in advanced non-small cell lung cancer.

Abstract

Background: The microRNA (miR)-200 family is implicated in regulating the immune checkpoint protein programmed death-ligand 1 (PD-L1), a key factor in lung cancer progression and response to immunotherapy. This study investigates the relationship between miR-200 expression and PD-L1 in non-small cell lung cancer (NSCLC), aiming to clarify its potential as a prognostic biomarker and a therapeutic target in immune checkpoint inhibitor (ICI) treatment for NSCLC.

Methods: RNA sequencing (RNA-seq) data from public databases were analyzed for correlation between miR-200 family expression and PD-L1 levels in lung cancer. MiR-200 and PD-L1 expression were assessed in lung cancer cell lines by reverse transcription quantitative polymerase chain reaction (RT-qPCR) and flow cytometry, respectively. To evaluate functional impact, miR-200 mimics were transfected into cell lines, and PD-L1 protein levels were measured. The influence of interferon gamma (IFN-γ) on miR-200 and PD-L1 expressions in cell lines were examined using RT-qPCR and flow cytometry. Serum samples and tumor biopsies were collected from advanced NSCLC patients before ICI therapy. Serum miR-200a was quantified by Droplet digital polymerase chain reaction (ddPCR), and its correlation with tumor PD-L1 and progression-free survival (PFS) was analyzed.

Results: Analysis of RNA-seq data revealed a significant inverse correlation between miR-200 family expression and PD-L1 levels in lung cancer (P<0.001). This was corroborated in cell lines, where miR-200a and miR-200b levels were significantly higher in low-PD-L1 cells compared to high-PD-L1 cells (P=0.01 and P=0.003). MiR-200a mimic transfection significantly decreased PD-L1 protein in H1975 and OKa-C-1 cells (P<0.001). IFN-γ stimulation increased PD-L1 expression but did not alter miR-200 levels. In advanced NSCLC patients, low serum miR-200a was associated with higher tumor PD-L1 expression (P=0.042) and significantly prolonged PFS following ICI therapy (median PFS: miR-200a-high, 129 days vs. miR-200a-low, 200 days; P=0.008).

Conclusions: This study shows that miR-200a regulates PD-L1 expression in NSCLC, affecting immune evasion. Serum miR-200a levels could serve as a non-invasive biomarker to predict PD-L1 expression and immunotherapy outcomes, helping identify patients who may benefit. Modulating miR-200a may also offer a new strategy to reduce PD-L1 in tumors, enhancing immune response.