DNA damage is overcome by TRIP13 overexpression during cisplatin nephrotoxicity
Taketsugu Hama, Prashanth K.B. Nagesh, Pallabita Chowdhury, Bob M. Moore II, Murali M. Yallapu, Kevin R. Regner, Frank Park
Taketsugu Hama, Prashanth K.B. Nagesh, Pallabita Chowdhury, Bob M. Moore II, Murali M. Yallapu, Kevin R. Regner, Frank Park
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Research Article Nephrology

DNA damage is overcome by TRIP13 overexpression during cisplatin nephrotoxicity

Abstract

Cisplatin is a commonly used chemotherapeutic agent to treat a wide array of cancers that is frequently associated with toxic injury to the kidney due to oxidative DNA damage and perturbations in cell cycle progression leading to cell death. In this study, we investigated whether thyroid receptor interacting protein 13 (TRIP13) plays a central role in the protection of the tubular epithelia following cisplatin treatment by circumventing DNA damage. Following cisplatin treatment, double-stranded DNA repair pathways were inhibited using selective blockers to proteins involved in either homologous recombination or non-homologous end joining. This led to increased blood markers of acute kidney injury (AKI) (creatinine and neutrophil gelatinase–associated lipocalin), tubular damage, activation of DNA damage marker (γ-H2AX), elevated appearance of G2/M blockade (phosphorylated histone H3 Ser10 and cyclin B1), and apoptosis (cleaved caspase-3). Conditional proximal tubule–expressing Trip13 mice were observed to be virtually protected from the cisplatin nephrotoxicity by restoring most of the pathological phenotypes back toward normal conditions. Our findings suggest that TRIP13 could circumvent DNA damage in the proximal tubules during cisplatin injury and that TRIP13 may constitute a new therapeutic target in protecting the kidney from nephrotoxicants and reduce outcomes leading to AKI.

Authors

Taketsugu Hama, Prashanth K.B. Nagesh, Pallabita Chowdhury, Bob M. Moore II, Murali M. Yallapu, Kevin R. Regner, Frank Park

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Figure 2

Trip13 overexpression in the proximal tubules provides protection from renal tubular epithelial cell damage following cisplatin administration.

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Trip13 overexpression in the proximal tubules provides protection from ...
Mice were injected with a single injection of vehicle (20% captisol in saline) or cisplatin (15 mg/kg IP), and kidneys and blood were harvested to detect markers of renal dysfunction. (A) Kidney samples from homozygous floxed stop (Trip13Stop/Stop) and Trip13ΔStop mice were harvested from mice treated with either vehicle (-Cis; 20% captisol) or cisplatin (+Cis; 15 mg/kg IP) after 72 hours following the initial injection. Western blot analysis was performed using FLAG and GFP primary antibodies. GAPDH was used as a loading control. Arrows on the right side of the panel indicate protein standard size. (B) Serum creatinine and (C) NGAL was measured in each mouse group. (D) Percentage of tubular damage was determined in each group. (E–H) Representative histological images from (E and G) vehicle- or (F and H) cisplatin-treated (15 mg/kg IP) Trip13Stop/Stop (E and F) and Trip13ΔStop (G and H) mice. Tubular epithelial cell damage was scored as a percentage of total tubules counted. — = vehicle; + = cisplatin. *P < 0.05, ****P < 0.0001 between all other groups using 1-way ANOVA with Tukey’s post hoc analysis. n = 4–8 animals per group. Scale bar: 200 μm (E–H).