What Carries the Signal in Pathology Foundation-Model Atlases? A Patient-Level Controlled Benchmark in Breast Cancer

Foundation model embeddings of screening mammograms may encode pre-diagnostic tissue change without task-specific adaptation. We tested whether embeddings move faster along a data-derived "cancer direction" in women later biopsied for cancer than in matched screen-negative controls, and whether this depends on pretraining domain. We studied 1,773 biopsied women (785 malignant, 988 biopsy-negative) and 1,773 matched controls, each with at least two annual screening exams before their index exam. An identical pipeline was applied to four 2D models: Mammo-CLIP (MC, out-of-distribution mammography), HOPPR (in-distribution mammography), MedImageInsight (MII, general medical imaging), and BiomedCLIP (biomedical vision-language pretraining on literature figures). Breast-level embeddings quantified longitudinal movement along the cancer direction. We compared cases and controls using a between-patient design with complementary mixed-effects analysis, and biopsied versus healthy contralateral breasts within patients. Under matched modality in MII embedding space, malignant cases drifted significantly faster than controls in the first two screening intervals preceding the index exam; biopsy-negative cases showed significance only in the first. MC differences were significant in the first interval for both biopsy groups. Within-patient comparisons showed a broadly similar pattern, with MC significance extending to the second interval in both groups and HOPPR showing significance at interval 1. BiomedCLIP showed no significant differences in either design or biopsy group. Overall, directional embedding velocity emerges as a property of clinically grounded rather than general biomedical pretraining, showing that foundation model embeddings can encode pre-diagnostic mammographic change without task-specific adaptation.

Graph neural networks (GNNs) are increasingly used to model biological systems, yet the reliability of post-hoc explanation methods for recovering meaningful molecular mechanisms remains unclear. Here, we systematically evaluate four widely used approaches: Saliency Attribution (SA), Integrated Gradients (IG), GNNExplainer, and Layer-wise Relevance Propagation (LRP) for identifying disease-relevant structure in breast cancer RNA-seq data projected onto a protein-protein interaction network. Using synthetic benchmarks with known ground-truth motifs, we show that explanation methods recover distinct signal organizations: SA performs best for sparse single-node drivers, whereas IG and LRP preferentially recover distributed pathway-like and cascade-like signals. In TCGA BRCA data, we identify a consistent topological signature of disease-associated hubs in which attribution peaks in the immediate 1-hop neighborhood and decays across successive network shells, a pattern most pronounced for IG and LRP and associated with strong enrichment of known cancer hubs. We further observe a trade-off between local hub enrichment and global gene ranking performance, with IG optimizing local enrichment and SA achieving superior global discrimination. Motivated by these complementary behaviors, we introduce a framework combining a shell-based hub score with consensus ranking across explainers. Consensus scores improve prioritization of canonical cancer genes (TP53, BRCA1, ESR1, MYC), reduce dependence on node degree, and, especially when tuned, outperform individual methods. Pathway enrichment further reveals improved recovery of biologically coherent cancer programs, including ERBB2, RTK, MAPK, immune, and cytokine signaling. Together, these results demonstrate that topology-aware integration of graph explanations can improve biological interpretability and biologically relevant molecular recovery.

H&E-stained whole-slide images offer cohort-scale availability and rich spatial context but lack molecular specificity, whereas bulk RNA-seq provides transcriptome-wide resolution at high cost with limited archival availability. We show that training a lightweight alignment module atop frozen histopathology and RNA-Seq foundation models enables open-vocabulary molecular prompting -- querying H&E slides with gene-set signatures to predict pathway activity without sequencing or end-to-end retraining. Using contrastive learning on a multi-cancer cohort (N=1,720), we achieve a 25-fold improvement in retrieval over baseline methods. Systematic analysis reveals a graduated predictability spectrum: morphologically grounded programs (cell-cycle programs, immune-related) are most reliably predicted (R^2>0.5), while predicting pathways with no morphological footprint remains challenging as expected. We validate clinical utility on the POSEIDON clinical trial: H&E-predicted squamous cell carcinoma scores recapitulate NSCLC subtype identity and predicted IFN-gamma mirror PD-L1 tumor-cell expression groups. Furthermore, genesets describing immune activation and fibrosis predict known tumor microenvironment archetypes from histology alone. We further validate generalization of our approach across unseen cohorts and demonstrate data-efficient domain adaptation, establishing a slide-native framework for molecular analysis on H&E images.