Fibroblast expression of transmembrane protein smoothened governs microenvironment characteristics after acute kidney injury
Yuan Gui, Haiyan Fu, Zachary Palanza, Jianling Tao, Yi-Han Lin, Wenjian Min, Yi Qiao, Christopher Bonin, Geneva Hargis, Yuanyuan Wang, Peng Yang, Donald L. Kreutzer, Yanlin Wang, Yansheng Liu, Yanbao Yu, Youhua Liu, Dong Zhou
Yuan Gui, Haiyan Fu, Zachary Palanza, Jianling Tao, Yi-Han Lin, Wenjian Min, Yi Qiao, Christopher Bonin, Geneva Hargis, Yuanyuan Wang, Peng Yang, Donald L. Kreutzer, Yanlin Wang, Yansheng Liu, Yanbao Yu, Youhua Liu, Dong Zhou
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Research Article Nephrology

Fibroblast expression of transmembrane protein smoothened governs microenvironment characteristics after acute kidney injury

Abstract

The smoothened (Smo) receptor facilitates hedgehog signaling between kidney fibroblasts and tubules during acute kidney injury (AKI). Tubule-derived hedgehog is protective in AKI, but the role of fibroblast-selective Smo is unclear. Here, we report that Smo-specific ablation in fibroblasts reduced tubular cell apoptosis and inflammation, enhanced perivascular mesenchymal cell activities, and preserved kidney function after AKI. Global proteomics of these kidneys identified extracellular matrix proteins, and nidogen-1 glycoprotein in particular, as key response markers to AKI. Intriguingly, Smo was bound to nidogen-1 in cells, suggesting that loss of Smo could affect nidogen-1 accessibility. Phosphoproteomics revealed that the ‘AKI protector’ Wnt signaling pathway was activated in these kidneys. Mechanistically, nidogen-1 interacted with integrin β1 to induce Wnt in tubules to mitigate AKI. Altogether, our results support that fibroblast-selective Smo dictates AKI fate through cell-matrix interactions, including nidogen-1, and offers a robust resource and path to further dissect AKI pathogenesis.

Authors

Yuan Gui, Haiyan Fu, Zachary Palanza, Jianling Tao, Yi-Han Lin, Wenjian Min, Yi Qiao, Christopher Bonin, Geneva Hargis, Yuanyuan Wang, Peng Yang, Donald L. Kreutzer, Yanlin Wang, Yansheng Liu, Yanbao Yu, Youhua Liu, Dong Zhou

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Figure 6

Specific deletion of Smo in fibroblasts mitigates AKI induced by cisplatin.

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Specific deletion of Smo in fibroblasts mitigates AKI induced by cisplat...
(A) Schematic diagram. i.p., intraperitoneal. At 3 days after cisplatin (Cis) injection, (B) Serum creatinine (Scr) levels in Gli1-Smo+/+ and Gli1-Smo–/– mice (n = 12). (C) Scr levels in Pdgfr-β-Smo+/+ and Pdgfr-β-Smo–/– mice (n = 12). (D and E) Western blots assay showing NGAL and Bax protein levels in Gli1-Smo+/+ and Gli1-Smo–/– (D) or in Pdgfr-β-Smo+/+ and Pdgfr-β-Smo–/– (E) kidneys. (F) Periodic Acid–Schiff (PAS) staining showing kidney morphological changes in Gli1-Smo+/+ and Gli1-Smo–/– mice. Blue asterisks indicate injured tubules. Representative micrographs of TUNEL assay or IHC staining against CD45 in Gli1-Smo+/+ and Gli1-Smo–/– kidneys. Scale bar: 25 μm. White arrows indicate apoptotic cells adn black arrows indicate CD45+ cells. (G) Representative micrographs of PAS staining, TUNEL assay, and IHC staining against CD45 in Pdgfr-β-Smo+/+ and Pdgfr-β-Smo–/– kidneys. Scale bar: 25 μm. Blue asterisks indicate injured tubules. White and black arrows, respectively, indicate apoptotic cells and CD45+ cells. (H and I) Western blots assay of NID1, Wnt1, and integrin β1 proteins in Gli1-Smo+/+ and Gli1-Smo–/– (H) or in Pdgfr-β-Smo+/+ and Pdgfr-β-Smo–/– (I) kidneys. Quantitative data are accordingly presented in J and K (Sham, n = 3; Cis, n = 5). †P < 0.05 versus sham control, *P < 0.05 versus Gli1-Smo+/+ or Pdgfr-β-Smo+/+ mice. Dots indicate individual animals in a given group. (L) IHC staining showing NID1, Wnt1, and integrin β1 (Intg β1) in Gli1-Smo+/+ and Gli1-Smo–/– or in Pdgfr-β-Smo+/+ and Pdgfr-β-Smo–/– kidneys. Scale bar: 25 μm. Arrows indicate positive cells. DAPI is a nuclear counterstain. For all Western blot panels, numbers indicate individual animals in a given group. Graphs are presented as means ± SEM. Differences among groups were analyzed using 1-way ANOVA, followed by the Student-Newman-Keuls test.