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Abstract: TH-PO441

TNF-Treated Kidney Organoids Recapitulate Poor Outcome Nephrotic Syndrome

Session Information

Category: Glomerular Diseases

  • 1302 Glomerular Diseases: Immunology and Inflammation

Authors

  • Fischer, Matthew, University of Michigan Michigan Medicine, Ann Arbor, Michigan, United States
  • Lassé, Moritz, Universitatsklinikum Hamburg-Eppendorf Zentrum fur Innere Medizin, Hamburg, Hamburg, Germany
  • Eddy, Sean, University of Michigan Michigan Medicine, Ann Arbor, Michigan, United States
  • El Saghir, Jamal, University of Michigan Michigan Medicine, Ann Arbor, Michigan, United States
  • Vega-Warner, V., University of Michigan Michigan Medicine, Ann Arbor, Michigan, United States
  • Kretzler, Matthias, University of Michigan Michigan Medicine, Ann Arbor, Michigan, United States
  • Rinschen, Markus M., Aarhus Universitet, Aarhus, Midtjylland, Denmark
  • Harder, Jennifer L., University of Michigan Michigan Medicine, Ann Arbor, Michigan, United States
Background

Nephrotic syndrome (NS) includes heterogeneous rare diseases (i.e. FSGS and MCD) complicating therapeutic development and intervention; individuals with NS could benefit greatly from a precision medicine-based approach. Human pluripotent stem cell derived kidney organoids (hKOs) provide an opportunity to identify pathomechanisms and therapeutics pertinent to subgroups of NS patients. We recently identified 3 subgroups of proteinuric individuals in NEPTUNE (1 with poorer outcomes) based on kidney tissue transcription profiles. Differential gene expression and pathway analysis of the poor outcome subgroup revealed a 272-gene signature of TNF activation, a pattern also seen in TNF-treated hKOs. The aim of this study was to further evaluate alterations induced in TNF-treated hKOs to assess their ability to capture the TNF-associated disease state.

Methods

hKOs were treated with TNF up to 48h. hKO lysates and culture media were analyzed for changes in gene and protein expression by a combination of qRT-PCR, ELISA, IF, proteomics, bulk and single cell RNAseq. Genes reflecting the TNF-induced hKO response were then evaluated in the NEPTUNE cohort tissue based bulk and single nuclear RNAseq datasets.

Results

Proteomic analysis of hKOs identified 320 peptides differentially expressed by TNF treatment, an overlap of 1.7% with the 272-gene TNF signature. Interrogation of the NEPTUNE cohort subgroups revealed that summary expression of genes encoding this peptide set was significantly increased in the poor outcome subgroup. Further evaluation of proteins secreted in TNF-treated hKO cultures revealed increased C3 and VCAM, both in the original 272-gene TNF signature. Expression of C3 and VCAM1 was higher in kidney tubular cells in NEPTUNE individuals with high TNF activation scores.

Conclusion

We discovered a set of peptides from TNF-treated hKOs that captured disease relevant molecular events in poor outcome NS. Further, we found that genes encoding TNF-induced proteins secreted by hKOs were also expressed by tubular cells in individuals with poor outcome NS. Our findings demonstrate the ability of the TNF-treated kidney organoid model to identify pathomechanistic elements relevant to a subset of individuals with nephrotic syndrome and to aid in discovery of non-invasive biomarkers.

Funding

  • Other NIH Support