Abstract: SA-PO606
IL233 Regulates Mitochondrial Function and WNT Signaling for Lupus Glomerulonephritis Remission
Session Information
- Glomerular Diseases: Immunology, Inflammation - II
November 09, 2019 | Location: Exhibit Hall, Walter E. Washington Convention Center
Abstract Time: 10:00 AM - 12:00 PM
Category: Glomerular Diseases
- 1202 Glomerular Diseases: Immunology and Inflammation
Authors
- Venkatadri, Rajkumar, Division of Nephrology, Charlottesville, Virginia, United States
- Sabapathy, Vikram, Division of Nephrology, Charlottesville, Virginia, United States
- Dogan, Murat, Division of Nephrology, Charlottesville, Virginia, United States
- Mohammad, Saleh, Division of Nephrology, Charlottesville, Virginia, United States
- Fu, Shu man, Division of Rheumatology, Charlottesville, Virginia, United States
- Sharma, Rahul, Division of Nephrology, Charlottesville, Virginia, United States
Background
We showed that the hybrid cytokine IL233 induced persistent remission in ongoing lupus glomerulonephritis (GN) in NZM2328 mice. The progression of GN in NZM2328 animals involves stages of acute (aGN), transitional (tGN) and chronic GN (cGN). As a means to further understand the mechanisms involved in IL233-rendered protection, we investigated modulation of mitochondrial function and canonical Wnt signaling that is understudied in the setting of aGN to cGN progression, utilizing both in vitro and in vivo approaches.
Methods
Mouse glomerular endothelial cells (mGECs) and proximal tubular TKPTS cells were treated with varying concentrations of IL233 and investigated for mitochondrial membrane potential and for genes and proteins associated with mitochondrial function. In vivo, kidney lysates from treated NZM2328 animals were screened for transcripts of mitochondrial genes by real time PCR. Canonical Wnt signaling proteins were studied by Western blotting. Seahorse XF Cell Mito Stress assay was performed with isolated regulatory T cells (Tregs) from treated animals.
Results
IL233 treatment induced a marked elevation of genes related to mitochondrial function and biogenesis (Pgc1α, Mfn1, Nrf1, Nrf2, Tfam and Drp1) in mGECs and TKPTS cells indicating a direct regulation of mitochondrial function. These cells also displayed higher mitochondrial membrane potential by flow cytometry. IL233-treated mice with established GN had higher expression of genes related to mitochondrial function and biogenesis, thus, complementing in vitro observations. Levels of canonical Wnt activators (LRP6 and Dvl3) and ��-catenin were significantly reduced in IL233 treated kidneys. Tregs isolated from saline, IL-2, IL-33, IL2+IL-33 and IL233 treated animals were investigated for oxidative respiration with Seahorse. Although there was a trend of a higher rate of oxidative respiration in mice treated with IL-2 or IL-33 only or the combination of IL-2 and IL-33, the Tregs isolated from IL233 treated mice had a significantly higher basal and burst respiration rate.
Conclusion
We present evidence for the use of IL233 for therapy of lupus GN in a murine model. In vitro and in vivo data reveals enhancement of mitochondrial function and modulation of canonical Wnt signaling as possible mechanisms of IL233-mediated remission from lupus GN.
Funding
- NIDDK Support