Abstract: TH-PO1074
A Multi-Omics Approach to Renal Epithelial Senescence Urinary Biomarker Discovery
Session Information
- CKD Progression and Complications: Diagnosis, Prognosis, Risk Factors
November 02, 2023 | Location: Exhibit Hall, Pennsylvania Convention Center
Abstract Time: 10:00 AM - 12:00 PM
Category: CKD (Non-Dialysis)
- 2303 CKD (Non-Dialysis): Mechanisms
Authors
- Baird, David Paul, The University of Edinburgh Centre for Inflammation Research, Edinburgh, Edinburgh, United Kingdom
- Reck, Maximilian, The University of Edinburgh The Queen's Medical Research Institute, Edinburgh, Edinburgh, United Kingdom
- Campbell, Ross Alexander, The University of Edinburgh Centre for Inflammation Research, Edinburgh, Edinburgh, United Kingdom
- O Sullivan, Eoin D., The University of Edinburgh Centre for Inflammation Research, Edinburgh, Edinburgh, United Kingdom
- Docherty, Marie, The University of Edinburgh Centre for Inflammation Research, Edinburgh, Edinburgh, United Kingdom
- Traynor, Jamie P., University of Glasgow, Glasgow, Glasgow, United Kingdom
- Mark, Patrick Barry, University of Glasgow, Glasgow, Glasgow, United Kingdom
- Hughes, Jeremy, The University of Edinburgh The Queen's Medical Research Institute, Edinburgh, Edinburgh, United Kingdom
- Denby, Laura, The University of Edinburgh The Queen's Medical Research Institute, Edinburgh, Edinburgh, United Kingdom
- Conway, Bryan, The University of Edinburgh The Queen's Medical Research Institute, Edinburgh, Edinburgh, United Kingdom
- Mylonas, Katie J., The University of Edinburgh Centre for Inflammation Research, Edinburgh, Edinburgh, United Kingdom
- Ferenbach, David A., The University of Edinburgh Centre for Inflammation Research, Edinburgh, Edinburgh, United Kingdom
Group or Team Name
- Ferenbach.
Background
Epithelial senescence is proposed as a driver of kidney fibrosis with senescent cell (SC) depletion improving outcomes. There are no non-invasive biomarkers for quantifying renal SCs. We used samples from patients with kidney disease and cultured human renal proximal tubular epithelial cells (hRPTECs) to identify urinary biomarkers of renal tubular senescence (rtSCs).
Methods
hRPTEC senescence was induced in vitro using 10Gy irradiation. Bulk RNAseq was performed comparing SCs with proliferating controls (n=5/group).
Immunofluorescence staining for p21CIP1, Ki67 and tubular markers CD10/CKPAN was performed in human kidney tissue from 131 CKD patients. P21CIP1 (+) / Ki67 (neg) tubular cells were classified as senescent (expressed as % of all tubular cells). In subgroup 1 (collected in Edinburgh, n=51), LC-MS studies were performed on matched urine samples. Proteins qualified as candidate biomarkers if they predicted the level of histological senescence in multivariate linear regression models alongside baseline eGFR, age and ACR and were upregulated in senescence transcriptomically. Candidate biomarkers were validated in subgroup 2 (collected in Glasgow, matching baseline characteristics to subgroup 1, n=53) that had both urine and kidney tissue available.
Results
In vitro: Irradiation increased mRNA levels of CDKN1A and reduced LMNB1 and MKI67 in keeping with senescence induction.
In vivo: rtSCs increased with age (rho = 0.58, p<0.001) and inversely with baseline eGFR (rho = -0.48, p<0.001). 331 proteins were detected by LC-MS. 5 candidate biomarkers were identified; 3 of which remained highly correlated with and predictive of histological senescence in the validation subgroup (fig. 1) [not named pending patent applications].
Conclusion
We have identified and validated 3 urinary biomarkers of senescence. These could aid patient selection for clinical trials of senolytic treatments in kidney disease.
Spearman rank correlation used.
Funding
- Government Support – Non-U.S.