Kidney Week

Abstract: SA-OR18

Role of Tcf21 in Mediating Renal Stromal Cell Fate Determination: Insights from Single-Cell Transcriptomics

Session Information

• Bioengineered Model Systems and Insights in Kidney Development and Function
  October 26, 2024 | Location: Room 2, Convention Center
  Abstract Time: 05:50 PM - 06:00 PM

Category: Development, Stem Cells, and Regenerative Medicine

• 600 Development, Stem Cells, and Regenerative Medicine

Authors

• Finer, Gal, Division of Pediatric Nephrology, Ann and Robert H Lurie Children's Hospital of Chicago, Chicago, Illinois, United States

Gal Finer, MD, PhD

• Zhou, Yalu, Feinberg Cardiovascular and Renal Research Institute, Northwestern University Feinberg School of Medicine, Chicago, Illinois, United States

Yalu Zhou, MS, MSc

• Khan, Mohammad Daud, Division of Rheumatology, Northwestern University Feinberg School of Medicine, Chicago, Illinois, United States

Mohammad Daud Khan, MS

• Gadhvi, Gaurav, Division of Rheumatology, Northwestern University Feinberg School of Medicine, Chicago, Illinois, United States

Gaurav Gadhvi, MS

• Yacu, George S., Division of Pediatric Nephrology, Ann and Robert H Lurie Children's Hospital of Chicago, Chicago, Illinois, United States

George S. Yacu

• Park, Joo-Seop, Feinberg Cardiovascular and Renal Research Institute, Northwestern University Feinberg School of Medicine, Chicago, Illinois, United States

Joo-Seop Park, PhD

• Gomez, Roberto Ariel, Department of Pediatrics, Child Health Research Center, University of Virginia, Charlottesville, Virginia, United States

Roberto Ariel Gomez, MD

• Sequeira Lopez, Maria Luisa S., Department of Pediatrics, Child Health Research Center, University of Virginia, Charlottesville, Virginia, United States

Maria Luisa S. Sequeira Lopez, MD

• Quaggin, Susan E., Feinberg Cardiovascular and Renal Research Institute, Northwestern University Feinberg School of Medicine, Chicago, Illinois, United States

Susan E. Quaggin, MD, FASN

• Winter, Deborah R., Division of Rheumatology, Northwestern University Feinberg School of Medicine, Chicago, Illinois, United States

Deborah R. Winter, PhD

Background

Normal kidney development requires coordinated interactions between multiple progenitor cell lineages. The Foxd1+ stromal progenitors are critical for normal nephrogenesis and their heterogeneity is increasingly appreciated. However, the molecular mechanisms and trajectories that drive the differentiation of Foxd1+ cells into renal stroma, capsule, mesangial cells, renin cells, pericytes, and vascular smooth muscle cells are poorly understood. Our previous work demonstrated that deletion of the mesoderm-specific bHLH transcription factor Tcf21 from Foxd1+ cells leads to a significant reduction in medullary stroma size and perivascular cell number. Here we investigate the role of Tcf21 in kidney stromal cell fate determination.

Methods

We isolated single cells from kidneys of Foxd1^Cre/+;Rosa26^mTmG/+;Tcf21^f/f cKO and Foxd1^Cre/+;Rosa26^mTmG/+;Tcf21^+/+ control mice at E14.5. Following enrichment for GFP+ cells by flow cytometry sorting, cDNA library was prepared using the 10x Genomics Chromium Platform and sequenced at a depth of 50,000 reads per cell.

Results

Clustering of the entire dataset (n=32,461) identified a large stromal population and a smaller representation of non-stromal lineages. Sub-clustering of stromal cells (n=22,355) identified seven molecularly distinct populations: medullary/perivascular, proliferating, ECM-expressing, differentiating nephron, nephrogenic, collecting duct, and ureteric-associated stroma. Loss of Tcf21 resulted in a dramatic reduction in medullary/perivascular, proliferating, nephrogenic, and collecting duct associated stroma. Lineage tracing of Foxd1 indicated a severe reduction in the medullary stromal space from E14.5 through E18.5. A novel population, expressing elevated levels of extracellular matrix components, was exclusively present in Tcf21 cKO kidneys. DEG analysis demonstrated that members of the Tgfβ/SMAD signaling were among the upregulated genes in Tcf21 cKO.

Conclusion

Taken together, these data underscore a role for Tcf21 in the emergence of the milieu of Foxd1+ derivatives; loss of Tcf21 leads to a shift in stromal cell fates that results in abnormal kidney development.

Funding

• NIDDK Support