Indoxyl Sulfate Inhibits Osteogenesis in Bone Marrow Mesenchymal Stem Cells through the AhR/Hes1 Pathway
Abstract
:1. Introduction
2. Results
2.1. IS Suppresses the Mineralization of D1 Cells
2.2. IS Downregulates the Expression of Osteogenic Markers in D1 Cells
2.3. IS Activates AhR Signaling
2.4. IS Impairs Osteogenesis by Inducing AhR
2.5. IS Upregulates the Expression of Hes1 through AhR
2.6. Hes1 Downregulates the Expression of Runx2 and Bmp2
2.7. Hes1 Directly Binds to the Promoters of Runx2 and Bmp2
3. Discussion
4. Materials and Methods
4.1. Cell Culture and Treatment
4.2. Alizarin Red S Staining
4.3. Quantitative Real-Time Polymerase Chain Reaction
4.4. Western Blotting
4.5. Measurement of Cytoplasmic and Nuclear AhR Concentrations
4.6. Immunofluorescence Staining
4.7. Ahr Knockdown
4.8. Luciferase Assay
4.9. Chromatin Immunoprecipitation
4.10. Statistical Analysis
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
- GBD Chronic Kidney Disease Collaboration. Global, regional, and national burden of chronic kidney disease, 1990–2017: A systematic analysis for the Global Burden of Disease Study 2017. Lancet 2020, 395, 709–733. [Google Scholar] [CrossRef] [PubMed]
- Naylor, K.L.; McArthur, E.; Leslie, W.D.; Fraser, L.A.; Jamal, S.A.; Cadarette, S.M.; Pouget, J.G.; Lok, C.E.; Hodsman, A.B.; Adachi, J.D.; et al. The three-year incidence of fracture in chronic kidney disease. Kidney Int. 2014, 86, 810–818. [Google Scholar] [CrossRef] [PubMed]
- Sprague, S.M.; Bellorin-Font, E.; Jorgetti, V.; Carvalho, A.B.; Malluche, H.H.; Ferreira, A.; D’Haese, P.C.; Drueke, T.B.; Du, H.; Manley, T.; et al. Diagnostic Accuracy of Bone Turnover Markers and Bone Histology in Patients with CKD Treated by Dialysis. Am. J. Kidney Dis. 2016, 67, 559–566. [Google Scholar] [CrossRef] [PubMed]
- El-Husseini, A.; Abdalbary, M.; Lima, F.; Issa, M.; Ahmed, M.T.; Winkler, M.; Srour, H.; Davenport, D.; Wang, G.; Faugere, M.C.; et al. Low Turnover Renal Osteodystrophy With Abnormal Bone Quality and Vascular Calcification in Patients with Mild-to-Moderate CKD. Kidney Int. Rep. 2022, 7, 1016–1026. [Google Scholar] [CrossRef] [PubMed]
- Goto, S.; Fujii, H.; Hamada, Y.; Yoshiya, K.; Fukagawa, M. Association between indoxyl sulfate and skeletal resistance in hemodialysis patients. Ther. Apher. Dial. 2010, 14, 417–423. [Google Scholar] [CrossRef] [PubMed]
- Iwasaki-Ishizuka, Y.; Yamato, H.; Nii-Kono, T.; Kurokawa, K.; Fukagawa, M. Downregulation of parathyroid hormone receptor gene expression and osteoblastic dysfunction associated with skeletal resistance to parathyroid hormone in a rat model of renal failure with low turnover bone. Nephrol. Dial. Transplant. 2005, 20, 1904–1911. [Google Scholar] [CrossRef]
- Kim, Y.H.; Kwak, K.A.; Gil, H.W.; Song, H.Y.; Hong, S.Y. Indoxyl sulfate promotes apoptosis in cultured osteoblast cells. BMC Pharmacol. Toxicol. 2013, 14, 60. [Google Scholar] [CrossRef] [PubMed]
- Idziak, M.; Pedzisz, P.; Burdzinska, A.; Gala, K.; Paczek, L. Uremic toxins impair human bone marrow-derived mesenchymal stem cells functionality in vitro. Exp. Toxicol. Pathol. 2014, 66, 187–194. [Google Scholar] [CrossRef]
- Duranton, F.; Cohen, G.; De Smet, R.; Rodriguez, M.; Jankowski, J.; Vanholder, R.; Argiles, A.; European Uremic Toxin Work, G. Normal and pathologic concentrations of uremic toxins. J. Am. Soc. Nephrol. 2012, 23, 1258–1270. [Google Scholar] [CrossRef]
- Stevens, E.A.; Mezrich, J.D.; Bradfield, C.A. The aryl hydrocarbon receptor: A perspective on potential roles in the immune system. Immunology 2009, 127, 299–311. [Google Scholar] [CrossRef]
- Gonzalez, F.J.; Fernandez-Salguero, P. The aryl hydrocarbon receptor: Studies using the AHR-null mice. Drug Metab. Dispos. 1998, 26, 1194–1198. [Google Scholar]
- Huang, B.; Butler, R.; Miao, Y.; Dai, Y.; Wu, W.; Su, W.; Fujii-Kuriyama, Y.; Warner, M.; Gustafsson, J.A. Dysregulation of Notch and ERalpha signaling in AhR-/- male mice. Proc. Natl. Acad. Sci. USA 2016, 113, 11883–11888. [Google Scholar] [CrossRef]
- Gerard, D.; Schmidt, F.; Ginolhac, A.; Schmitz, M.; Halder, R.; Ebert, P.; Schulz, M.H.; Sauter, T.; Sinkkonen, L. Temporal enhancer profiling of parallel lineages identifies AHR and GLIS1 as regulators of mesenchymal multipotency. Nucleic Acids Res. 2019, 47, 1141–1163. [Google Scholar] [CrossRef] [PubMed]
- Ryan, E.P.; Holz, J.D.; Mulcahey, M.; Sheu, T.J.; Gasiewicz, T.A.; Puzas, J.E. Environmental toxicants may modulate osteoblast differentiation by a mechanism involving the aryl hydrocarbon receptor. J. Bone Miner. Res. 2007, 22, 1571–1580. [Google Scholar] [CrossRef]
- Schroeder, J.C.; Dinatale, B.C.; Murray, I.A.; Flaveny, C.A.; Liu, Q.; Laurenzana, E.M.; Lin, J.M.; Strom, S.C.; Omiecinski, C.J.; Amin, S.; et al. The uremic toxin 3-indoxyl sulfate is a potent endogenous agonist for the human aryl hydrocarbon receptor. Biochemistry 2010, 49, 393–400. [Google Scholar] [CrossRef]
- Liu, W.C.; Shyu, J.F.; Lin, Y.F.; Chiu, H.W.; Lim, P.S.; Lu, C.L.; Zheng, C.M.; Hou, Y.C.; Chen, P.H.; Lu, K.C. Resveratrol Rescue Indoxyl Sulfate-Induced Deterioration of Osteoblastogenesis via the Aryl Hydrocarbon Receptor/MAPK Pathway. Int. J. Mol. Sci. 2020, 21, 7483. [Google Scholar] [CrossRef] [PubMed]
- Kageyama, R.; Ohtsuka, T.; Kobayashi, T. The Hes gene family: Repressors and oscillators that orchestrate embryogenesis. Development 2007, 134, 1243–1251. [Google Scholar] [CrossRef] [PubMed]
- Vecera, J.; Prochazkova, J.; Sumberova, V.; Panska, V.; Paculova, H.; Lanova, M.K.; Masek, J.; Bohaciakova, D.; Andersson, E.R.; Pachernik, J. Hypoxia/Hif1alpha prevents premature neuronal differentiation of neural stem cells through the activation of Hes1. Stem Cell Res. 2020, 45, 101770. [Google Scholar] [CrossRef]
- Grogan, S.P.; Olee, T.; Hiraoka, K.; Lotz, M.K. Repression of chondrogenesis through binding of notch signaling proteins HES-1 and HEY-1 to N-box domains in the COL2A1 enhancer site. Arthritis Rheum. 2008, 58, 2754–2763. [Google Scholar] [CrossRef]
- Hilton, M.J.; Tu, X.; Wu, X.; Bai, S.; Zhao, H.; Kobayashi, T.; Kronenberg, H.M.; Teitelbaum, S.L.; Ross, F.P.; Kopan, R.; et al. Notch signaling maintains bone marrow mesenchymal progenitors by suppressing osteoblast differentiation. Nat. Med. 2008, 14, 306–314. [Google Scholar] [CrossRef]
- Nobta, M.; Tsukazaki, T.; Shibata, Y.; Xin, C.; Moriishi, T.; Sakano, S.; Shindo, H.; Yamaguchi, A. Critical regulation of bone morphogenetic protein-induced osteoblastic differentiation by Delta1/Jagged1-activated Notch1 signaling. J. Biol. Chem. 2005, 280, 15842–15848. [Google Scholar] [CrossRef] [PubMed]
- Thomsen, J.S.; Kietz, S.; Strom, A.; Gustafsson, J.A. HES-1, a novel target gene for the aryl hydrocarbon receptor. Mol. Pharmacol. 2004, 65, 165–171. [Google Scholar] [CrossRef]
- Faust, D.; Vondracek, J.; Krcmar, P.; Smerdova, L.; Prochazkova, J.; Hruba, E.; Hulinkova, P.; Kaina, B.; Dietrich, C.; Machala, M. AhR-mediated changes in global gene expression in rat liver progenitor cells. Arch. Toxicol. 2013, 87, 681–698. [Google Scholar] [CrossRef]
- Dong, B.; Cheng, W.; Li, W.; Zheng, J.; Wu, D.; Matsumura, F.; Vogel, C.F. FRET analysis of protein tyrosine kinase c-Src activation mediated via aryl hydrocarbon receptor. Biochim. Biophys. Acta 2011, 1810, 427–431. [Google Scholar] [CrossRef] [PubMed]
- Naruse, M.; Ishihara, Y.; Miyagawa-Tomita, S.; Koyama, A.; Hagiwara, H. 3-Methylcholanthrene, which binds to the arylhydrocarbon receptor, inhibits proliferation and differentiation of osteoblasts in vitro and ossification in vivo. Endocrinology 2002, 143, 3575–3581. [Google Scholar] [CrossRef] [PubMed]
- Ronis, M.J.; Watt, J.; Pulliam, C.F.; Williams, A.E.; Alund, A.W.; Haque, E.; Gadupudi, G.S.; Robertson, L.W. Skeletal toxicity resulting from exposure of growing male rats to coplanar PCB 126 is associated with disruption of calcium homeostasis and the GH-IGF-1 axis and direct effects on bone formation. Arch. Toxicol. 2020, 94, 389–399. [Google Scholar] [CrossRef] [PubMed]
- Tong, Y.; Niu, M.; Du, Y.; Mei, W.; Cao, W.; Dou, Y.; Yu, H.; Du, X.; Yuan, H.; Zhao, W. Aryl hydrocarbon receptor suppresses the osteogenesis of mesenchymal stem cells in collagen-induced arthritic mice through the inhibition of beta-catenin. Exp. Cell Res. 2017, 350, 349–357. [Google Scholar] [CrossRef] [PubMed]
- Ge, L.; Cui, Y.; Cheng, K.; Han, J. Isopsoralen Enhanced Osteogenesis by Targeting AhR/ERalpha. Molecules 2018, 23. [Google Scholar] [CrossRef]
- An, L.; Shi, Q.; Fan, M.; Huang, G.; Zhu, M.; Zhang, M.; Liu, Y.; Weng, Y. Benzo[a]pyrene injures BMP2-induced osteogenic differentiation of mesenchymal stem cells through AhR reducing BMPRII. Ecotoxicol. Environ. Saf. 2020, 203, 110930. [Google Scholar] [CrossRef]
- Pfeuty, B. A computational model for the coordination of neural progenitor self-renewal and differentiation through Hes1 dynamics. Development 2015, 142, 477–485. [Google Scholar] [CrossRef]
- Watanabe, N.; Tezuka, Y.; Matsuno, K.; Miyatani, S.; Morimura, N.; Yasuda, M.; Fujimaki, R.; Kuroda, K.; Hiraki, Y.; Hozumi, N.; et al. Suppression of differentiation and proliferation of early chondrogenic cells by Notch. J. Bone Miner. Metab. 2003, 21, 344–352. [Google Scholar] [CrossRef]
- Zayzafoon, M.; Abdulkadir, S.A.; McDonald, J.M. Notch signaling and ERK activation are important for the osteomimetic properties of prostate cancer bone metastatic cell lines. J. Biol. Chem. 2004, 279, 3662–3670. [Google Scholar] [CrossRef] [PubMed]
- Deregowski, V.; Gazzerro, E.; Priest, L.; Rydziel, S.; Canalis, E. Notch 1 overexpression inhibits osteoblastogenesis by suppressing Wnt/beta-catenin but not bone morphogenetic protein signaling. J. Biol. Chem. 2006, 281, 6203–6210. [Google Scholar] [CrossRef] [PubMed]
- Zanotti, S.; Smerdel-Ramoya, A.; Canalis, E. HES1 (hairy and enhancer of split 1) is a determinant of bone mass. J. Biol. Chem. 2011, 286, 2648–2657. [Google Scholar] [CrossRef]
- Locklin, R.M.; Riggs, B.L.; Hicok, K.C.; Horton, H.F.; Byrne, M.C.; Khosla, S. Assessment of gene regulation by bone morphogenetic protein 2 in human marrow stromal cells using gene array technology. J. Bone Miner. Res. 2001, 16, 2192–2204. [Google Scholar] [CrossRef]
- de Jong, D.S.; Steegenga, W.T.; Hendriks, J.M.; van Zoelen, E.J.; Olijve, W.; Dechering, K.J. Regulation of Notch signaling genes during BMP2-induced differentiation of osteoblast precursor cells. Biochem. Biophys. Res. Commun. 2004, 320, 100–107. [Google Scholar] [CrossRef]
- Lo, R.; Celius, T.; Forgacs, A.L.; Dere, E.; MacPherson, L.; Harper, P.; Zacharewski, T.; Matthews, J. Identification of aryl hydrocarbon receptor binding targets in mouse hepatic tissue treated with 2,3,7,8-tetrachlorodibenzo-p-dioxin. Toxicol. Appl. Pharmacol. 2011, 257, 38–47. [Google Scholar] [CrossRef]
- Sciaudone, M.; Gazzerro, E.; Priest, L.; Delany, A.M.; Canalis, E. Notch 1 impairs osteoblastic cell differentiation. Endocrinology 2003, 144, 5631–5639. [Google Scholar] [CrossRef] [PubMed]
- Shindo, K.; Kawashima, N.; Sakamoto, K.; Yamaguchi, A.; Umezawa, A.; Takagi, M.; Katsube, K.; Suda, H. Osteogenic differentiation of the mesenchymal progenitor cells, Kusa is suppressed by Notch signaling. Exp. Cell Res. 2003, 290, 370–380. [Google Scholar] [CrossRef]
- Wang, W.; Liu, X.; Wang, W.; Li, J.; Li, Y.; Li, L.; Wang, S.; Zhang, J.; Zhang, Y.; Huang, H. The Effects of Indoxyl Sulfate on Human Umbilical Cord-Derived Mesenchymal Stem Cells In Vitro. Cell Physiol. Biochem. 2016, 38, 401–414. [Google Scholar] [CrossRef]
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Hsieh, C.-W.; Chang, L.-H.; Wang, Y.-H.; Li, W.-T.; Chang, J.-K.; Chen, C.-H.; Ho, M.-L. Indoxyl Sulfate Inhibits Osteogenesis in Bone Marrow Mesenchymal Stem Cells through the AhR/Hes1 Pathway. Int. J. Mol. Sci. 2024, 25, 8770. https://doi.org/10.3390/ijms25168770
Hsieh C-W, Chang L-H, Wang Y-H, Li W-T, Chang J-K, Chen C-H, Ho M-L. Indoxyl Sulfate Inhibits Osteogenesis in Bone Marrow Mesenchymal Stem Cells through the AhR/Hes1 Pathway. International Journal of Molecular Sciences. 2024; 25(16):8770. https://doi.org/10.3390/ijms25168770
Chicago/Turabian StyleHsieh, Chin-Wen, Ling-Hua Chang, Yan-Hsiung Wang, Wei-Ting Li, Je-Ken Chang, Chung-Hwan Chen, and Mei-Ling Ho. 2024. "Indoxyl Sulfate Inhibits Osteogenesis in Bone Marrow Mesenchymal Stem Cells through the AhR/Hes1 Pathway" International Journal of Molecular Sciences 25, no. 16: 8770. https://doi.org/10.3390/ijms25168770