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New insights into cyclosporine A nephrotoxicity by proteome analysis
Abstract:
Using two-dimensional gel electrophoresis (2-DE), we recently discovered an association between decreased calcium-binding protein, calbindin-D 28 kDa, urinary calcium wasting and intratubular corticomedullary calcifications in rat kidney. This observation prompted us to investigate kidney tissues of other species, including man. In this paper we show that in dogs and monkeys, which are generally devoid of cyclosporine A (CsA)-mediated nephrotoxicity, renal calbindin levels were not affected by the CsA treatment whereas in CsA-treated human kidney-transplant recipients with renal vascular or tubular toxicity, a marked decrease in renal calbindin-D 28 kDa protein level was found in most of the kidney biopsy sections. The present results strongly suggest that calbindin is a marker for CsA-nephrotoxicity. The discovery of calbindin-D 28 kDa being involved in CsA toxicity has evolved from the application of 2-DE and has not been reported previously, proving that proteomics can provide essential information in mechanistic toxicology. Considering the current improvements in proteome methods it is expected that high throughput proteomics will become an indispensable tool in preclinical safety testing.
Insights
Calbindin-D 28 kDa, a calcium-binding protein, is decreased in cyclosporine A (CsA)-induced kidney toxicity. This finding suggests calbindin is a potential biomarker for CsA nephrotoxicity in humans.
Area of Science:
- Nephrology
- Toxicology
- Proteomics
Background:
- Cyclosporine A (CsA) is a widely used immunosuppressant with known nephrotoxic potential.
- Previous studies identified a link between decreased calbindin-D 28 kDa, urinary calcium wasting, and calcifications in rat kidneys.
Purpose of the Study:
- To investigate the role of calbindin-D 28 kDa as a potential biomarker for CsA-induced nephrotoxicity across different species.
- To determine if CsA treatment affects renal calbindin levels in species resistant to its nephrotoxic effects.
Main Methods:
- Two-dimensional gel electrophoresis (2-DE) was used to identify protein changes.
- Analysis of kidney tissues from CsA-treated and untreated dogs, monkeys, and human kidney-transplant recipients.
- Assessment of renal calbindin-D 28 kDa protein levels in biopsy sections.
Main Results:
- In dogs and monkeys, CsA treatment did not alter renal calbindin levels.
- CsA-treated human kidney-transplant recipients with evidence of nephrotoxicity showed a significant decrease in renal calbindin-D 28 kDa protein.
- Calbindin-D 28 kDa levels were unaffected in CsA-treated animals lacking nephrotoxicity.
Conclusions:
- Calbindin-D 28 kDa is a promising biomarker for detecting cyclosporine A-induced nephrotoxicity in humans.
- Proteomics, specifically 2-DE, offers valuable insights into drug-induced toxicity mechanisms.
- High-throughput proteomics may become essential for preclinical safety assessments.