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Abstract: SA-PO1066

Metabolic Changes in Peripheral Blood Mononuclear Cells Isolated from Dialysis Patients with Vascular Access Dysfunction

Session Information

Category: Pharmacology (PharmacoKinetics, -Dynamics, -Genomics)

  • 1800 Pharmacology (PharmacoKinetics, -Dynamics, -Genomics)

Authors

  • Altintas, Mehmet M., Rush University Medical Center, Chicago, Illinois, United States
  • DiBartolo, Salvatore, Rush University Medical Center, Chicago, Illinois, United States
  • Samelko, Beata, Rush University Medical Center, Chicago, Illinois, United States
  • Wasse, Monnie, Rush University Medical Center, Chicago, Illinois, United States
Background

As numerous complex pathologies can stem from cellular energy dysfunction, we aimed to elucidate whether mitochondrial dysfunction contributes to arteriovenous fistula (AVF) and arteriovenous graft (AVG) failure in a cohort of dialysis patients. We used peripheral blood mononuclear cells (PBMCs) as the model system of disease monitoring for bioenergetic analyses.

Methods

The bioenergetics study was conducted using PBMCs and serum from dialysis patients with AVF or AVG (re)stenosis. PBMCs and serum are isolated from whole blood through the use of a density gradient centrifugation, aliquoted and frozen at -80°C until analysis. On the day of analysis, PBMCs from healthy controls and patients were thawed, diluted and counted before being seeded into Seahorse XF24 assay plate to detect changes in mitochondrial respiration. The bioenergetics analysis was performed in the presence of Seahorse XF medium (free of bicarbonate, pH 7.4) using mitochondrial stress test kit and Seahorse flux analyzer. In order to test the metabolic changes caused by patient serum, we used commercially available control PBMCs and treated those with 10% serum from healthy controls and patients in 6-well plates. After 24 hours, cells were harvested and loaded to Seahorse XF24 assay plates for the bioenergetic analysis.

Results

We developed a technique to measure mitochondrial oxygen consumption in PBMCs isolated from dialysis patients with AVF or AVG (re)stenosis and control PBMCs fed with patient’s serum for 24 h. In PBMCs of patients, we found a reduction in each of fundamental parameters of mitochondrial function such as basal respiration, ATP turnover, proton leak, maximal respiration and spare respiratory capacity. A similar trend was observed when the control PBMCs were cultured in the presence of 10% serum from patients for 24 h.

Conclusion

Our data demonstrates a correlation between mitochondrial oxygen consumption of PBMCs and end-stage renal disease (ESRD) in a case-control study of 30 patients. We propose a link between mitochondrial dysfunction and vascular access failure since PBMCs are exposed to metabolic and hemodynamic stimuli in the vasculature. Our findings and the methodology may help identify individuals at risk for hemodialysis vascular access dysfunction.

Funding

  • NIDDK Support