Kidney Week

Abstract: TH-PO096

Hedgehog Interacting Protein (Hhip) Deficiency in Endothelial Cells Prevents Ischemia Reperfusion-Induced Renal Tubular Cell Injury Through Inhibition of NF-KB Signaling

Session Information

• AKI: Mechanisms - I
  November 02, 2023 | Location: Exhibit Hall, Pennsylvania Convention Center
  Abstract Time: 10:00 AM - 12:00 PM

Category: Acute Kidney Injury

• 103 AKI: Mechanisms

Authors

• Pang, Yuchao, Centre de Recherche du Centre Hospitalier de l'Universite de Montreal, Montreal, Quebec, Canada

Yuchao Pang,

• Peng, Junzheng, Centre de Recherche du Centre Hospitalier de l'Universite de Montreal, Montreal, Quebec, Canada

Junzheng Peng,

• Zhao, Xin-Ping, Centre de Recherche du Centre Hospitalier de l'Universite de Montreal, Montreal, Quebec, Canada

Xin-Ping Zhao,

• Chang, Shiao-Ying, Centre de Recherche du Centre Hospitalier de l'Universite de Montreal, Montreal, Quebec, Canada

Shiao-Ying Chang,

• Liao, Min-Chun, Centre de Recherche du Centre Hospitalier de l'Universite de Montreal, Montreal, Quebec, Canada

Min-Chun Liao,

• Rivard, Alain, Centre de Recherche du Centre Hospitalier de l'Universite de Montreal, Montreal, Quebec, Canada

Alain Rivard,

• Ingelfinger, Julie R., Massachusetts General Hospital, Boston, Massachusetts, United States

Julie R. Ingelfinger,

• Chan, John S.D., Centre de Recherche du Centre Hospitalier de l'Universite de Montreal, Montreal, Quebec, Canada

John S.D. Chan,

• Zhang, Shao-Ling, Centre de Recherche du Centre Hospitalier de l'Universite de Montreal, Montreal, Quebec, Canada

Shao-Ling Zhang,

Background

Acute kidney injury (AKI) caused by renal ischemia reperfusion (R I/R) is a major clinical issue that is lacking viable therapies, and its underlying mechanisms are poorly understood. Given the important role of hedgehog interacting protein in chronic kidney disease-related renal fibrosis, we investigated whether Hhip knockout in endothelial cells (EC) (Hhip^EC KO) could prevent R I/R-induced kidney injury in vivo and in vitro.

Methods

Male Hhip^EC KO and control Hhip^fl/fl mice at 10-weeks old were subjected to 45 minutes of the unilateral R I/R or sham surgery, both with contralateral nephrectomy. The AKI time-course was examined at Day1, Day 3, Day7 and Day 21. In vitro, naïve renal proximal tubular cells (IRPTCs) exposed to the conditioned media harvested from mouse endothelial cell (mECs) with or without Hhip (siRNA) ± gentamicin treatment, were studied.

Results

Compared to the respective controls, R I/R mice (Hhip^fl/fl and Hhip^EC KO) body weight loss (BW, g) occurred from Day 1 until Day 21, regardless of Hhip deficiency. In line with the AKI time course, Hhip^fi/fl-R I/R mice displayed kidney dysfunction marked by increased urinary albumin-to-creatinine ratio; tubular/endothelial cell injury and fibrosis as assessed by kidney morphological staining (Periodic Acid-Schiff, Masson’s trichrome and Sirius Red) with enhanced cystatin C-, kidney-induced molecular 1-and TGFβ1- immunoexpression in their kidneys. Maladaptive and inadequate renal tubular damage repair was observed by enhanced vascular cell adhesion protein 1 and NF/kB (p50/p65) expression. In contrast, these changes were significantly ameliorated in the kidneys of Hhip^EC KO -R I/R mice. In vitro, naïve IRPTCs exposed to the conditioned media harvested from mECs treated with gentamicin (2.5mg/ml) had lower wound healing ability/capacity, higher cellular senescent activity and increased NF/kB expression. Those changes were largely prevented in naïve IRPTCs exposed to conditioned media harvested from Hhip KO mECs treated with gentamicin.

Conclusion

Our data suggest that Hhip knockout in endothelial cells prevented AKI-induced renal tubular injury. This action might be mediated, at least, in part, by inhibition of NF-KB signaling.

Funding

• Government Support – Non-U.S.