Securing diagonal integration of multimodal single-cell data against ambiguous mapping

Recent advances in single-cell multimodal omics technologies enable the exploration of cellular systems at unprecedented resolution, leading to the rapid generation of multimodal datasets that require sophisticated integration methods. Diagonal integration has emerged as a flexible solution for integrating heterogeneous single-cell data without relying on shared cells or features. However, the absence of anchoring elements introduces the risk of artificial integrations, where cells across modalities are incorrectly aligned due to ambiguous mapping. To address this challenge, we propose SONATA, a novel diagnostic method designed to detect potential artificial integrations resulting from ambiguous mappings in diagonal data integration. SONATA identifies ambiguous alignments by quantifying cell-cell ambiguity within the data manifold, ensuring that biologically meaningful integrations are distinguished from spurious ones. It is worth noting that SONATA is not designed to replace any existing pipelines for diagonal data integration; instead, SONATA works simply as an add-on to an existing pipeline for achieving more reliable integration. Through comprehensive evaluation on both simulated and real multimodal single-cell datasets, we observe that artificial integrations in diagonal data integration are widespread yet surprisingly overlooked, occurring across all mainstream diagonal integration methods. We demonstrate SONATAs ability to safeguard against misleading integrations and provide actionable insights into potential integration failures across mainstream methods. Our approach offers a robust framework for ensuring the reliability and interpretability of multimodal single-cell data integration. 1