Functionally Guided Graph Learning for Robust Cross-Patient Cell-Type Annotation in Single-Cell RNA Sequencing.
Cross-patient cell-type annotation in single-cell RNA sequencing (scRNA-seq) remains challenging due to pronounced interpatient heterogeneity and distribution shifts across patient-specific cellular contexts. Conventional annotation approaches often rely on proximity-driven graph construction or expression similarity, which may introduce spurious cell-cell connections and lead to unstable knowledge transfer across patients. To address this limitation, we propose PathoGraph, a functionally guided graph learning framework for robust cross-patient cell-type annotation. The proposed method integrates KEGG-7-based biosemantic graph structure learning with cross-patient representation adaptation. Specifically, pathway-derived functional semantic profiles are incorporated to refine patient-specific cell graphs, encouraging biologically coherent neighborhoods and suppressing noise introduced by purely expression-based similarity. Based on the refined graphs, a cross-patient representation adaptation mechanism further aligns embeddings between labeled reference patients and unlabeled query patients to facilitate reliable annotation transfer. Experiments on three cross-patient scRNA-seq data sets, including leukemia, breast invasive carcinoma, and colorectal cancer data sets, demonstrate that PathoGraph achieves stable annotation performance across 32 directed reference-to-query transfer tasks. Across all tasks, PathoGraph obtained an average ACC of 84.28% and an F1-score of 84.08%, showing competitive and stable performance compared with representative marker-based, correlation-based, and model-based annotation methods. Ablation studies further show that removing the biosemantic graph learning module reduces the average accuracy to 83.48%, highlighting the importance of functional-guided graph refinement. In addition, post hoc functional relevance analyses in immune-cell and cancer-associated contexts suggest that the learned cell-cell graphs capture biologically relevant neighborhood structures beyond expression-driven proximity. The source code and processed data are publicly available at: https://github.com/LiYuechao1998/PathoGraph.