ASN's Mission

To create a world without kidney diseases, the ASN Alliance for Kidney Health elevates care by educating and informing, driving breakthroughs and innovation, and advocating for policies that create transformative changes in kidney medicine throughout the world.

learn more

Contact ASN

1401 H St, NW, Ste 900, Washington, DC 20005

email@asn-online.org

202-640-4660

The Latest on X

Kidney Week

Abstract: FR-PO192

Evaluating Proteomics in a Rat Model of AKI

Session Information

  • AKI: Mechanisms
    October 25, 2024 | Location: Exhibit Hall, Convention Center
    Abstract Time: 10:00 AM - 12:00 PM

Category: Acute Kidney Injury

  • 103 AKI: Mechanisms

Authors

  • Schwabauer, Denise L., Inotiv, Inc., Westminster, Colorado, United States
  • Camarillo, Jeannie M., Inotiv, Inc., Nashville, Tennessee, United States
  • Richards, Toni L., Inotiv, Inc., Westminster, Colorado, United States
  • Liebler, Daniel C., Inotiv, Inc., Nashville, Tennessee, United States
  • Minor, Kenneth H., Inotiv, Inc., Westminster, Colorado, United States
Background

Acute kidney injury (AKI) secondary to acute renal ischemia is associated with high mortality and morbidity, few effective treatments, and risk of chronic kidney disease. AKI therapeutics can be advanced with well characterized disease models, which inform early therapeutic development. Here we describe the proteomic landscape of a rat model of AKI and describe characteristic injury pathways and potential therapeutic targets.

Methods

Rats were subjected to sham or warm, bilateral renal ischemia (40’) using proprietary vascular clamps. Rats were housed in metabolic cages for 24 or 48 hours post-reperfusion. In 24 hour intervals, blood and urine samples were collected and evaluated for progression of kidney injury biomarkers. At each endpoint, renal tissue was harvested and subjected to global proteomic analysis using tandem mass tags (TMT) and high resolution mass spectrometry (MS) to define quantitative changes in the kidney proteome following renal ischemia/reperfusion (I/R). Gene set enrichment analysis (GSEA) was utilized to identify cellular pathways significantly altered between sham and I/R timepoints.

Results

Proteomic analysis identified 3,084 proteins that were altered across the I/R time course. GSEA identified several protein networks and pathways that displayed time-dependent alteration with development of AKI. Pathways significantly upregulated with injury include complement and coagulation cascades, cell adhesion and metabolic pathways, and kidney inflammation/injury-associated pathways. Proteomic alterations were correlated with data from orthogonal approaches including gene expression profiling and immunohistochemistry.

Conclusion

Proteomic profiling captures both well-characterized and novel molecular alterations underlying renal injury. Global proteomics enables quantitative assessment of changes in the kidney proteome in the progression of AKI. GSEA with rich proteome data enables rapid identification of cellular pathways significantly altered following renal I/R and represents all the features of the injury phenotype. Variations of the protocol may enable nuanced dissection of injury. Moreover, targeted MS provides robust biomarker assays for any protein, without antibodies. Distinct biomarker expression profiles of varying durations of renal I/R can identify potential therapeutic targets and possible new AKI diagnostic markers.

Funding

  • Commercial Support – Inotiv