Search bioRxiv⌕ Search

bioRxiv · 10.1101/2025.06.17.660070

Cell-Type Specific Single-Cell Signatures Reveal Nephrotoxic Drug Effects

Abstract

Drug-induced acute kidney injury (AKI) affects about 20% of hospitalized AKI patients, a significant contributor to morbidity and mortality. The lack of understanding of the kidney system and functioning of nephrotoxic drugs contributes to hospital-acquired AKI cases. AKI is difficult to predict because of its complex injury mechanism and the numerous pathways through which it manifests. Traditional toxicity biomarkers, like elevated creatinine levels, detect AKI only after significant kidney injury has occurred. Concurrently, advancements in single cell RNA sequencing (scRNAseq) have improved our ability to map cellular heterogeneity within tissues, potentially enabling the study of drug effects at a single cell level. We hypothesized that only particular subtypes of kidney cells may be responsible for observed nephrotoxicity and explain prediction challenges. To test this, we generated cellular response scores for 32 kidney cell types from the Human Cell Atlas and estimated drug effects. We identified significant expression differences in 6 cell types (e.g. Indistinct intercalated cell p = 0.009, Epithelial Progenitor cell, p = 0.04). We also developed an ensemble model that achieved an AUROC of 0.6 across different kidney cell populations - a significant improvement over using traditional bulk RNA sequencing alone. The single-cell transcriptomic signatures we identified potentially reveal unexplained molecular mechanisms of nephrotoxicity. Author SummaryThe prediction and early detection of drug-induced kidney injury is a significant clinical challenge since physicians rely on biomarkers that only become elevated after substantial kidney damage has occurred, limiting opportunities for intervention and patient protection. Our investigation utilized single-cell data and available drug toxicity information to examine how individual kidney cell populations respond to potentially harmful medications. We hypothesized that specific kidney cell subtypes are primarily responsible for observed drug toxicity, which may explain the difficulties in predicting drug-induced kidney injury. Through comprehensive analysis of 32 distinct kidney cell types, we identified six specific cellular populations that demonstrate differential responses to nephrotoxic compounds. We subsequently developed models that demonstrate superior predictive performance compared to analytical approaches using bulk RNA sequencing data. Our methodology represents a substantial advancement in precision medicine approaches to drug safety. These findings have important implications for clinical practice and patient safety. The cellular signatures we identified may enable earlier detection of kidney injury risk, potentially allowing clinicians to modify treatment regimens before irreversible damage occurs. Our work establishes a foundation for improved drug safety protocols and may contribute to reducing medication-related kidney injury in hospitalized patients.

Explore related subjects

Keep this discovery

Explore connections, maps & timelines

BibTeXRIS

Kuchi, A. S., Acitores Cortina, J. M., Liu, H., Fatapour, Y., Berkowitz, J., Tatonetti, N.. 2025-06-22. Cell-Type Specific Single-Cell Signatures Reveal Nephrotoxic Drug Effects. https://doi.org/10.1101/2025.06.17.660070

Cite the original work for its findings. Save a collection to share your selection of sources.

KEEP EXPLORING

Related preprints

Comparative functional genomics reveals distributed SERPINE1 like characteristics among Drosophila serpins

Serpins (serine protease inhibitors) regulate proteolytic activity through a suicide-substrate inhibitory mechanism, whereby the serpin covalently traps and inactivates their target protease. Serpins are broadly conserved across eukaryotes, but the evolutionary and functional relationships between human SERPINE1, and Drosophila Serpins in incompletely defined. To identify candidate Drosophila functional analogs of SERPINE1/PAI-1, a multi-omics framework was applied to thoroughly compare the serpins across species. The multi-level analysis integrated orthology prediction, protein sequence and structural characterization, phylogenetic reconstruction, bulk tissue transcriptomics, single-cell transcriptomics, and aging-associated expression analysis. Candidate serpins (Spn42Da, Spn42Db, Spn42Dc, Spn42Dd, Spn28B, and Spn28F) were evaluated using DIOPT, AlphaFold/ChimeraX structural comparisons, reactive center loop analyses, phylogenetic distances, GTEx, Human Protein Atlas, FlyAtlas2, Fly Cell Atlas, and Aging Fly Cell Atlas datasets. Comparative analyses demonstrated full-length sequence and reactive center loop (RCL) divergence despite preservation of the canonical serpin fold. Tissue, cell-type, and age-associated expression analysis revealed distinct candidate-specific patterns. Spn42Dd comparisons uncovered similarities to human SERPINE1 in sequence/structural and broad expression but did not represent the strongest overall cross-species expression correlations. Spn42Da and Spn42Db received the highest cross species integrated support. Quality check of the ranking confirmed that Spn42Da and Spn42Db were stable top candidates across orthology, sequence alignment, and expression domains. The multi-domain analysis was unable to identify a single definitive Drosophila serpin as the closest counterpart to human SERPINE1, indicating that cross-species similarity is distributed across multiple candidate serpins. This study therefore provides a quantitative method for prioritizing potential evolutionary and functional analogs across highly divergent species.

bioinformatics↗

CellChat Hotspot: A Targeted Approach to Cell Cell Communication Analysis With Spatial transcriptomics Validation.

Abstract. In recent years, single cell RNA sequencing (scRNA seq) has enabled the investigation of intercellular communication through Cell Cell Communication (CCC) tools such as CellChat. Despite their potential, these approaches remain challenging to interpret, particularly when global analyses generate complex interaction networks that may include biologically irrelevant signals. To address this limitation, we developed a framework termed CellChat Hotspot, designed to improve the biological interpretability of inferred communication networks. This approach integrates gene set enrichment analysis with a context driven filtering strategy, selecting both genes and cells based on their functional relevance prior to communication inference. Applied to stage II colon cancer data, CellChat Hotspot enabled the identification of condition-specific interaction patterns associated with recurrence. Non recurrent patients showed enrichment of immune related processes, whereas recurrent cases were characterized by pathways associated with invasion and metastasis. Notably, a loss of interaction in the HSPG pathway between CALB1+ and stromal cells was observed in recurrent patients, suggesting a potential disruption of cell matrix adhesion mechanisms. To verify the reliability of this finding, we applied the framework to spatial transcriptomics data, which confirmed the presence of the HSPG2 DAG1 interaction in non-recurrent tumors and its near absence in recurrent ones. Overall, this framework provides a biologically guided strategy for studying intercellular communication in complex systems such as the tumor microenvironment.

bioinformatics↗

DiffTernary: a Diffusion Model for Molecular Glue Ternary Complex Docking

Molecular glues (MGs) modulate protein function by inducing or stabilizing protein interactions. However, accurately modeling MG ternary complexes remains challenging. Here, we present DiffTernary, a diffusion-based docking method for the joint prediction of relative protein arrangement and MG binding pose. We introduce Ternary-PDB, a dataset of 76,702 experimentally determined ternary complexes curated from the Protein Data Bank using structural filters and a release cutoff aligned with AlphaFold 3 (AF3). When trained on this dataset, DiffTernary achieved acceptable protein-protein interaction (PPI) predictions (DockQ > 0.23) for 50.0% of MGBench complexes with a single prediction, compared with 51.1% for AF3. In comparisons with AF3, OpenFold3, ESMFold2, Protenix-v1 and Protenix-v2, DiffTernary ranked second with a best-of-25 PPI prediction success rate of 78.4%, behind ESMFold2 (90.9%) and ahead of AF3 (72.7%). A separate DiffTernary model trained on TernaryDB achieved a mean DockQ of 0.306 on the 94 complexes in the TernaryDB MGD test set with a single prediction, compared with 0.228 for DeepTernary. Confidence-based selection among 40 samples yielded acceptable PPI predictions for 41.5% of complexes, compared with 30.9% for DeepTernary. An E2Former variant increased training and sampling throughput 6.90-fold and 4.37-fold, respectively, under the evaluated configurations. Together, DiffTernary and Ternary-PDB provide an efficient docking approach for MG complex prediction and a valuable structural resource for MG discovery and design.

bioinformatics↗