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Kidney Week

Abstract: FR-PO637

Selected Renal Cells Exhibit Renal Tubule Formation Associated with Transforming Growth Factor B2 Expression

Session Information

  • Pediatric Nephrology - II
    November 03, 2023 | Location: Exhibit Hall, Pennsylvania Convention Center
    Abstract Time: 10:00 AM - 12:00 PM

Category: Pediatric Nephrology

  • 1900 Pediatric Nephrology

Authors

  • Narayan, Prakash, ProKidney, Winston-Salem, North Carolina, United States
  • Bruce, Andrew T., ProKidney, Winston-Salem, North Carolina, United States
  • Rohlfing, Mark A., ProKidney, Winston-Salem, North Carolina, United States
  • Bertram, Tim A., ProKidney, Winston-Salem, North Carolina, United States
  • Jain, Deepak, ProKidney, Winston-Salem, North Carolina, United States
Background

TGFB2 mediates interactions between the ureteric bud and the metanephric mesenchyme resulting in renal tubule formation. Selected renal cells (SRCs) express ureteric bud markers RET and FGF8, and cap mesenchyme markers, LHX1, OSR1 and SIX2, their administration is therapeutic in models of chronic kidney disease. We tested the hypothesis that SRCs express tgfb2 and assemble into renal tubules.

Methods

Human SRCs (sourced from NDRI kidneys) were submitted to miRNAseq and mRNAseq, differentially expressed nodes identified and seeded into miRNet for interactome visualization. SRCs were submitted to scRNA-seq to map gene expression. SRCs were placed in culture, and secreted TGFB2 measured using ELISA, and evidence of renal tubule formation confirmed by antibody staining for epithelial markers GGT1 and SLC12A1.

Results

Compared to the source biopsy hsa-miR-145-5p (log2FC=-6.9) and hsa-miR-199a-5p (log2FC=-5.7) were downregulated in SRCs (padj<0.01). Gene ontology revealed that these miRNAs regulate expression of tgfb2 together with renal epithelial markers (A). SRCs overexpressed tgfb2 (log2FC=2.5, padj <0.01). scRNA-seq confirmed tgfb2 expression (B) and SRC secreted TGFB2 exhibited 12-fold increase vs. control; p<0.01). Gene ontology revealed that tgfb2 forms an interactome with ret, fgf8, lhx1, osr1 and six2, ureteric bud and cap mesenchyme markers expressed by SRCs (C). Placed in culture, SRCs assembled into GGT1- and SLC12A1-positive renal tubules (D).

Conclusion

These data suggest that therapeutic activity of SRCs may be mediated in part via formation of renal tubules and maintenance of electrolyte balance, fluid homeostasis, reabsorption of essential nutrients, urine concentration and Cystatin C metabolism.

(A) hsa-miR-145-and hsa-miR-199a-5p regulate expression of tgfb2 and renal epithelial markers. SRCs express tgfb2 (B) which forms an interactome with ureteric bud and cap mesenchyme markers (C). Placed in culture, SRCs assemble into GGT1-and SCL12A1-positive renal tubules (D, arrows).

Funding

  • Commercial Support – ProKidney