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Everolimus Alleviates Renal Allograft Interstitial Fibrosis by Inhibiting Epithelial to Mesenchymal Transition Not Only via Inducing Autophagy but Also via Stabilizing iκB-α

Z. Gui1, W. Zijie2, M. Zheng3, M. Gu1, R. Tan3

1Department of Urology, The Second Affiliated Hospital with Nanjing Medical University, Nanjing, China, 2Jiangsu Province Hospital, Nanjing, China, 3Department of Urology, The First Affiliated Hospital with Nanjing Medical University, Nanjing, China

Meeting: 2022 American Transplant Congress

Abstract number: 578

Keywords: Epithelial cells, Fibrosis, Kidney transplantation, Tumor necrosis factor (TNF)

Topic: Basic Science » Basic Clinical Science » 19 - Chronic Organ Rejection

Session Information

Session Name: Chronic Organ Rejection

Session Type: Rapid Fire Oral Abstract

Date: Tuesday, June 7, 2022

Session Time: 5:30pm-7:00pm

 Presentation Time: 6:40pm-6:50pm

Location: Hynes Room 309

*Purpose: Chronic allograft dysfunction (CAD) is the major cause of late graft loss in long-term renal transplantation. In our previous study, we found Epithelial-mesenchymal transition (EMT) is a significant event in the progression of renal allograft tubulointerstitial fibrosis, and impaired autophagic flux plays a critical role in renal allograft fibrosis. Everolimus (EVR) has been reported to widely used to prevent the progression of organs fibrosis and graft rejection. However, the pharmacological mechanism of EVR in kidney transplantation remains to be determined. The purpose of this study was to explore the effect and the mechanism of EVR on CAD after renal transplantation.

*Methods: We performed kidney transplantation of Balb/c mice kidneys into C57BL/6 recipients following bilateral nephrectomy to establish CAD mice model. Based on CAD models, we intervened with EVR to explore the effect of IGU on transplanted renal interstitial fibrosis. Hematoxylin-eosin (HE) staining, periodate schiff (PAS) staining, immunohistochemical and western blot detection were used to observe the pathological changes, level of autophagy and EMT of transplanted kidney in each group. In addition, human kidney 2 (HK2) cells line treated with tumor necrosis factor-α (TNF-α) and EVR on TNF-α-induced-EMT and autochondrial damage in vivo. Real-time fluorescence quantitative polymerase chain reaction and western blot were used to detect the related genes and proteins. RNA sequencing was used to detect the changes of gene transcriptome level after the treatment with TNF-α.

*Results: Here, we found that EVR could attenuate the progression of EMT and renal allograft interstitial fibrosis, and also activate autophagy in vivo. To explore the mechanism behind it, we detected the relationship among EVR, autophagy level and TNF-α-induced-EMT in HK2 cells. Our results showed that autophagy was upregulated upon mTOR pathway inhibition by EVR which could significant reduced expression of TNF-α-induced-EMT. However, the inhibition of EVR on TNF-α-induced-EMT was partly reversed following the addition of autophagy inhibitor chloroquine. In addition, we found that TNF-α activated EMT through protein kinase B (Akt) as well as nuclear factor kappa B (NF-κB) pathway according to the RNA sequencing, and EVR’s effect on the EMT was only associated with IκB-α stabilization instead of Akt pathway.

*Conclusions: Together, our findings suggest that EVR may retard impaired autophagic flux and block NF-κB pathway activation, thereby prevent progression of TNF-α-induced-EMT and renal allograft interstitial fibrosis.

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To cite this abstract in AMA style:

Gui Z, Zijie W, Zheng M, Gu M, Tan R. Everolimus Alleviates Renal Allograft Interstitial Fibrosis by Inhibiting Epithelial to Mesenchymal Transition Not Only via Inducing Autophagy but Also via Stabilizing iκB-α [abstract]. Am J Transplant. 2022; 22 (suppl 3). https://atcmeetingabstracts.com/abstract/everolimus-alleviates-renal-allograft-interstitial-fibrosis-by-inhibiting-epithelial-to-mesenchymal-transition-not-only-via-inducing-autophagy-but-also-via-stabilizing-i%ce%bab-%ce%b1/. Accessed May 30, 2025.

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