Abstract: FR-PO1016

Caspase Inhibition during Cold Storage Improves Graft Function and Histology of Transplanted Kidneys

Session Information

Category: Transplantation

  • 1701 Transplantation: Basic and Experimental

Authors

  • Jain, Swati, UC Denver, Aurora, Colorado, United States
  • Nydam, Trevor L., University of Colorado Denver, Aurora, Colorado, United States
  • Plenter, Robert J., University of Colorado Denver, Aurora, Colorado, United States
  • Jani, Alkesh, University of Denver Colorado, Aurora, Colorado, United States
Background

Prolonged cold ischemia is a risk factor for delayed graft function (DGF) of kidney transplants, and is associated with caspase-3 mediated apoptotic tubular cell death. We hypothesized that treatment of a donor organ with the caspase inhibitor, QVD-OPh, prior to kidney transplantation would be associated with significantly reduced renal tubular epithelial cell (RTEC) apoptosis, histology and improved renal function post-transplant in a mouse kidney transplant model of DGF.

Methods

For in vitro studies, mouse RTECs were incubated with either DMSO or QVD-OPh during cold storage in saline followed by rewarming in RPMI media. For in vivo studies, donor kidneys from C57BL/6 mice were perfused with either cold saline, DMSO (vehicle), or QVD-OPh, recovered, stored in the same solution at 4°C for 60 minutes, and transplanted into syngeneic C57BL/6 recipients. RTEC apoptosis, histological changes and sCr were quantitated

Results

Tubular cells treated with the caspase inhibitor QVD-OPh had significantly reduced capsase-3 protein expression, caspase-3 activity, and apoptotic cell death vs saline or DMSO (vehicle) in a dose dependent manner. Treatment of donor kidneys with QVD-OPh significantly reduced sCr, and resulted in significantly less tubular cell apoptosis, brush border injury, tubular injury, cast formation, and tubule lumen dilation vs. DMSO and saline treated kidneys (Table 1).

Conclusion

Treatment of RTECs and donor kidneys with Q-VD-OPh significantly reduces apoptosis, histological injury and improves renal function in a mouse model of kidney transplantation. Caspase inhibition may be a useful strategy to prevent DGF and increase the donor pool.

in vitro (cold storage and rewarming) n=3
 SalineDMSOQVD-OPh
Caspase-3 protein++++++_
Caspase-3 activity0.149 ± 0.0030.139 ± 0.0070.029 ± 0.003*
Apoptotic cell death21.98 ± 1.0719.12 ± 1.319.53 ± 0.66*
in vivo (kidney transplant)
 Control (n=4)Saline (n=11)DMSO (n=7)QVD-OPh (n=7)
Serum creatinine (mg/dl)0.33 ± 0.022.71 ± 0.211.85 ± 0.290.99 ±0.41*
Apoptotic cell death/hpf0.5 ± 0.13.3 ± 0.242.78 ± 0.281.1 ± 0.19*
TUNEL assay0.02 ± 0.021.31 ± 0.150.79 ± 0.10.3 ± 0.09*
Brush Border Injury1.03 ± 0.023.5 ± 0.083.29 ± 0.142.5 ± 0.23*
Tubular Injury07.64 ± 0.484.46 ± 0.473.2 ± 0.52*
Cast01.88 ± 2.341.92 ± 0.210.8 ± 0.15*
Lumen area22.5 ± 9.062030 ± 143.61887 ± 180.51166 ± 201.2*

*p<0.05 vs. Saline & DMSO

Funding

  • Veterans Affairs Support