Intratumoral SPP1+BCL2A1+ Tumor-Associated Macrophages Predict Poor Response to PD1 Blockade

<b>Background/Objectives:</b> Immune checkpoint blockade (ICB) has emerged as a promising therapeutic option for hepatocellular carcinoma (HCC), yet reliable biomarkers to predict clinical outcomes remain limited. Tumor-associated macrophages (TAMs) are increasingly recognized as key regulators of the tumor immune microenvironment. <b>Methods:</b> We interrogated a publicly available HCC single-cell RNA sequencing (scRNA-seq) dataset to characterize intratumoral immune cell subpopulations. Through unsupervised clustering and gene signature analysis, we identified a distinct subset of <i>SPP1</i> (secreted phosphoprotein 1, also known as osteopontin) and BCL2A1 (Bcl-2-related protein A1) double-positive TAMs. Their abundance was quantified and associated with patient outcomes. Further independent HCC transcriptomic datasets with annotated PD1-based ICB response status were used for examination. <b>Results:</b> Across the discovery (GSE149614; <i>n</i> = 10) cohort, elevated expression of intratumoral <i>SPP1</i>⁺<i>BCL2A1</i>⁺ TAMs was identified in HCC. In the ICB datasets (GSE151530; <i>n</i> = 4), patients with high <i>SPP1</i>⁺<i>BCL2A1</i>⁺ TAM expression further exhibited significantly poorer responses to ICB therapy. Further, the validation cohort (GSE206325; <i>n</i> = 18) confirmed these findings accordingly. Notably, these TAMs were expressed thoroughly within the immunosuppressive T-cell microenvironment in non-responders but were distinctly expressed among the cytotoxic T-cell responses in responders. <b>Conclusions:</b> Our findings identify <i>SPP1</i>⁺<i>BCL2A1</i>⁺ TAMs as a poor prognostic biomarker in HCC patients undergoing ICB therapy. By promoting an immunosuppressive microenvironment, <i>SPP1⁺BCL2A1⁺</i> TAMs, which are survival-advantaged, may represent both a predictive marker and a potential therapeutic target to enhance the efficacy of immunotherapy.