Renal failure is a frequent complication of septicemia that contributes significantly to mortality, particularly in postoperative patients in the ICU. In the U.S. septicemia is the 12th leading cause of death in adults and the 9th leading cause of death in children 1-4 years of age. Current therapy is, for the most part, only supportive. Thus, the long-term objective of our research is to identify new therapeutic targets to treat sepsis-induced renal injury. In this proposal, we will describe a functional defect in the kidney that may represent a previously unrecognized vascular pathway involved in polymicrobial sepsis- induced renal injury using the most clinically relevant murine model, cecal ligation and puncture (CLP). Our preliminary data show that inhibition of inducible nitric oxide synthase, inhibition of caspases, or inhibition of p53 each can prevent CLP-induced renal failure in mice. We also discovered a dramatic loss in perfused cortical peritubular capillaries following CLP that is ameliorated by these three classes of inhibitors. This unexpected and novel finding of a CLP-induced vascular defect mediated by reactive nitrogen species (RNS), caspases, and p53 is the basis of our central hypothesis that in sepsis, RNS-initiated activation of caspases results in a decline in peritubular capillary perfusion that leads to RNS-dependent tubular epithelial injury and ultimately renal failure. We will use biochemical and intravital videomicroscopy techniques to assess peritubular capillary dysfunction and tubular injury following mild and severe CLP in mice.
Aim 1 will determine if peritubular endothelial injury and capillary dysfunction are early events that precede tubule epithelial cell injury.
Aim 2 will determine the relative contributions of caspase-3 and p53 in peritubular capillary dysfunction and tubule epithelial cell injury using both pharmacological and genetic approaches.
Aim 3 will determine if RNS-induced activation of p53 and caspases contribute to CLP-induced peritubular capillary dysfunction and tubule epithelial cell injury using pharmacological a genetic approaches.
Both Aim 2 and Aim 3 will use in vivo studies as well as complementary in vitro studies with primary cultures of renal endothelial and tubular epithelial cells.

Public Health Relevance

These studies will supply valuable information on the cascade of signaling and cell- specific events that result in sepsis-induced renal injury. They will identify new signaling pathways and test them as new therapeutic targets for this devastating disease.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
5R01DK075991-05
Application #
8289664
Study Section
Surgery, Anesthesiology and Trauma Study Section (SAT)
Program Officer
Hoshizaki, Deborah K
Project Start
2008-08-01
Project End
2014-05-31
Budget Start
2012-06-01
Budget End
2014-05-31
Support Year
5
Fiscal Year
2012
Total Cost
$299,907
Indirect Cost
$91,636
Name
University of Arkansas for Medical Sciences
Department
Pharmacology
Type
Schools of Medicine
DUNS #
122452563
City
Little Rock
State
AR
Country
United States
Zip Code
72205
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Holthoff, Joseph H; Wang, Zhen; Patil, Naeem K et al. (2013) Rolipram improves renal perfusion and function during sepsis in the mouse. J Pharmacol Exp Ther 347:357-64
Mayeux, Philip R; MacMillan-Crow, Lee Ann (2012) Pharmacological targets in the renal peritubular microenvironment: implications for therapy for sepsis-induced acute kidney injury. Pharmacol Ther 134:139-55
Pathak, Elina; MacMillan-Crow, Lee Ann; Mayeux, Philip R (2012) Role of mitochondrial oxidants in an in vitro model of sepsis-induced renal injury. J Pharmacol Exp Ther 340:192-201
Holthoff, Joseph H; Wang, Zhen; Seely, Kathryn A et al. (2012) Resveratrol improves renal microcirculation, protects the tubular epithelium, and prolongs survival in a mouse model of sepsis-induced acute kidney injury. Kidney Int 81:370-8
Wang, Zhen; Holthoff, Joseph H; Seely, Kathryn A et al. (2012) Development of oxidative stress in the peritubular capillary microenvironment mediates sepsis-induced renal microcirculatory failure and acute kidney injury. Am J Pathol 180:505-16
Wang, Zhen; Herzog, Christian; Kaushal, Gur P et al. (2011) Actinonin, a meprin A inhibitor, protects the renal microcirculation during sepsis. Shock 35:141-7
Seely, Kathryn A; Holthoff, Joseph H; Burns, Samuel T et al. (2011) Hemodynamic changes in the kidney in a pediatric rat model of sepsis-induced acute kidney injury. Am J Physiol Renal Physiol 301:F209-17
Holthoff, Joseph H; Woodling, Kellie A; Doerge, Daniel R et al. (2010) Resveratrol, a dietary polyphenolic phytoalexin, is a functional scavenger of peroxynitrite. Biochem Pharmacol 80:1260-5
Pathak, Elina; Mayeux, Philip R (2010) In vitro model of sepsis-induced renal epithelial reactive nitrogen species generation. Toxicol Sci 115:475-81