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Kidney Week

Abstract: SA-OR070

Tcf21 Preserves Stromal Cell Identity and Restrains Profibrotic Fate Adoption During Kidney Development

Session Information

Category: Development, Stem Cells, and Regenerative Medicine

  • 600 Development, Stem Cells, and Regenerative Medicine

Authors

  • Yacu, George Shawket, Ann & Robert H Lurie Children's Hospital of Chicago, Chicago, Illinois, United States
  • Sayed, Mohammed, University of Cincinnati College of Medicine, Cincinnati, Ohio, United States
  • Zhou, Yalu, Northwestern University Feinberg School of Medicine, Chicago, Illinois, United States
  • Ward, Sarah E, Ann & Robert H Lurie Children's Hospital of Chicago, Chicago, Illinois, United States
  • Gomez, Roberto Ariel, University of Virginia School of Medicine, Charlottesville, Virginia, United States
  • Sequeira Lopez, Maria Luisa S., University of Virginia School of Medicine, Charlottesville, Virginia, United States
  • Park, Joo-Seop, Northwestern University Feinberg School of Medicine, Chicago, Illinois, United States
  • Lim, Hee Woong, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio, United States
  • Quaggin, Susan E., Northwestern Memorial Hospital, Chicago, Illinois, United States
  • Winter, Deborah R., Northwestern University Feinberg School of Medicine, Chicago, Illinois, United States
  • Finer, Gal, Ann & Robert H Lurie Children's Hospital of Chicago, Chicago, Illinois, United States
Background

Foxd1+ cells give rise to diverse cell types critical for nephron and vascular patterning, but the transcriptional control of their differentiation is not well understood. Tcf21 is a key regulator, though its downstream roles and long-term impact remain unclear.

Methods

We used scRNA-seq and scATAC-seq to profile Foxd1-lineage cells in E14.5 Tcf21-cKO and control kidneys. E18.5 transcriptomics, CellChat modeling, and immunostaining assessed inter-compartment effects. Adult kidney pathology and function were evaluated histologically and biochemically.

Results

scRNA-seq of E14.5 Foxd1-lineage cells revealed a spectrum of stromal subpopulations, including a mutant-specific Emcn+ population, absent in controls, enriched for ECM and fibrotic genes (Fras1, Postn, Sparcl1). scATAC-seq confirmed increased chromatin accessibility at the Emcn promoter and elevated Emcn gene activity in cKO. Mutant kidneys showed depletion of Collecting duct-associated, Nephrogenic zone-associated, Proliferating, and Medullary/Perivascular stroma, and a shift toward dysregulated differentiation. By E18.5, the Emcn+ population persisted, co-expressing Meis1/2/3 and matrisome genes, occupying perivascular zones and contributing to stromal expansion and vascular remodeling. Bulk RNA-seq revealed upregulation of fibrotic and ectopic muscle markers (Fap, Myh3, Inhba) and downregulation of epithelial genes (Aqp4, Slc14a1). CellChat analysis identified loss of developmental stromal cues (Bmp4-Bmpr1a+Acvr2b, Wnt5a-Fzd4) and emergence of maladaptive ligand-receptor signaling (Wnt4-Fzd4+Lrp6, Col4a5-Itga2+Itgb1). Immunostaining confirmed disrupted E-cad+ epithelium and endothelial disorganization. In adult mice, Tcf21-cKO kidneys exhibited persistent Emcn+ cells, interstitial fibrosis, glomerulosclerosis, reduced nephron number, hypodysplasia, and elevated serum creatinine, establishing a link between early stromal mis-specification and long-term fibrotic remodeling.

Conclusion

Tcf21 governs stromal cell identity during nephrogenesis, promoting key cell types while suppressing pro-fibrotic fates. Its loss induces aberrant Emcn+ cells, disrupts inter-compartment signaling, and leads to fibrosis and renal dysfunction, implicating it in the developmental origins of CKD.

Funding

  • NIDDK Support

Digital Object Identifier (DOI)