A statistical theory of graph regularization for RNA velocity near developmental bifurcations

Abstract

Graph-based regularization is widely used to stabilize noisy RNA-velocity estimates by encouraging transcriptionally similar cells to share similar velocity vectors. Near developmental bifurcations, however, proximity-based graphs may connect cells from distinct daughter lineages, reducing estimation variance at the cost of attenuating biologically meaningful lineage-specific dynamics. We formulate graph-regularized RNA velocity as a statistical estimation problem on a potentially misspecified cell-state graph and develop a theoretical framework for analyzing this trade-off. We derive an exact graph-spectral bias--variance decomposition that characterizes how Laplacian regularization suppresses estimation noise while introducing systematic smoothing bias. To quantify lineage preservation, we introduce a branch-sensitive risk that separates within-lineage denoising from cross-lineage information leakage. We further show that persistent cross-branch connectivity can induce nonvanishing branch bias, implying that standard proximity-graph regularization may remain asymptotically inconsistent near developmental bifurcations. These results provide a mathematical foundation for understanding both the statistical benefits and the geometric limitations of graph regularization for RNA-velocity estimation.