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Kidney Week

Abstract: FR-PO984

4-(2-Aminoethyl) Benzenesulfonyl Fluoride Hydrochloride (AEBSF) Reduced Kidney Fibrosis in Part by Targeting CREB3L1 in Myofibroblasts

Session Information

  • CKD: Pathobiology - I
    November 04, 2022 | Location: Exhibit Hall, Orange County Convention Center‚ West Building
    Abstract Time: 10:00 AM - 12:00 PM

Category: CKD (Non-Dialysis)

  • 2203 CKD (Non-Dialysis): Mechanisms

Authors

  • Yamamoto, Ayaha, Osaka Daigaku, Suita, Osaka, Japan
  • Obana, Masanori, Osaka Daigaku, Suita, Osaka, Japan
  • Fujio, Yasushi, Osaka Daigaku, Suita, Osaka, Japan
Background

Since kidney fibrosis is a critical event for the onset of renal failure, novel therapeutic agents to prevent and/or mitigate kidney fibrosis are urgently needed. Previously, we found that cAMP responsible element binding protein 3-like 1 (CREB3L1), a transcription factor, exacerbated kidney fibrosis, using conventional knockout mice. However, it is unclear that pharmacological approach to inhibit CREB3L1 is useful for the treatment of kidney fibrosis. CREB3L1 is cleaved by site-1/2 protease, resulting in activation. In this study, we examined whether 4-(2-aminoethyl) benzenesulfonyl fluoride hydrochloride (AEBSF), a site-1 protease inhibitor, suppressed kidney fibrosis.

Methods

CREB3L1 expression in human and murine kidneys was examined by immunohistochemistry with anti-CREB3L1 and α-SMA antibodies. To examine the effects of CREB3L1 in myofibroblasts on kidney fibrosis, myofibroblast-specific CREB3L1 knockout (cKO) mice were subjected to unilateral ureteral obstruction (UUO). Day 7 after UUO, kidney fibrosis was examined by Sirius Red staining, hydroxyproline assay and immunofluorescence analysis. Cultured TGF-β1-stimulated NRK49F cells (myofibroblasts) were treated with AEBSF. In addition, C57BL/6 mice were intraperitoneally injected with AEBSF for 9 consecutive days starting 2 days before UUO.

Results

CREB3L1 was increased in myofibroblasts in human and murine fibrotic kidneys. Kidney fibrosis was attenuated in CREB3L1 cKO mice compared with control mice. In addition, CREB3L1 cKO mice showed reduced number of Ki-67-positive proliferative myofibroblasts in fibrotic kidneys, suggesting that CREB3L1 in myofibroblasts may be therapeutic target for kidney fibrosis. AEBSF suppressed CREB3L1 activation in myofibroblasts. Significantly, AEBSF treatment reduced kidney fibrosis after UUO(hydroxyproline content: PBS-contralateral; 0.17±0.03 µg/mg, PBS-UUO; 0.32±0.06 µg/mg, AEBSF-contralateral; 0.12±0.03 µg/mg, AEBSF-UUO; 0.25±0.05 µg/mg, n = 9 for PBS, n = 6 for AEBSF).

Conclusion

AEBSF could be a novel therapeutic tool for kidney fibrosis by targeting CREB3L1 in myofibroblasts.