Urine-Derived Stem Cell-Secreted Klotho Plays a Crucial Role in the HK-2 Fibrosis Model by Inhibiting the TGF-β Signaling Pathway
Abstract
:1. Introduction
2. Results
2.1. Characterization and Homing Effect of UDSCs
2.2. Klotho Expression Patterns of UDSCs
2.3. TGF-β1 Induced HK-2 Fibrosis Model
2.4. Klotho siRNA Transfection Efficiency in UDSCs
2.5. UDSCs Suppress Fibrosis in HK-2 Cells via the TGF-β Signaling Pathway
3. Discussion
4. Materials and Methods
4.1. Isolation and Culture
4.2. Characterization
4.3. Adipogenic and Osteogenic Differentiation
4.4. Chondrogenic Differentiation
4.5. Homing Assay
4.6. Cell Culture
4.7. siRNA Assay
4.8. Co-Culture Assay
4.9. Real-Time PCR
4.10. Western Blotting
4.11. Klotho ELISA Assay
4.12. Immunofluorescence Staining
4.13. Statistical Analysis
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
- Zelmer, J.L. The economic burden of end-stage renal disease in Canada. Kidney Int. 2007, 72, 1122–1129. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Chow, F.Y.; Briganti, E.M.; Kerr, P.G.; Chadban, S.J.; Zimmet, P.Z.; Atkins, R.C. Health-related quality of life in Australian adults with renal insufficiency: A population-based study. Am. J. Kidney Dis. 2003, 41, 596–604. [Google Scholar] [CrossRef] [PubMed]
- Kusek, J.W.; Greene, P.; Wang, S.R.; Beck, G.; West, D.; Jamerson, K.; Agodoa, L.Y.; Faulkner, M.; Level, B. Cross-sectional study of health-related quality of life in African Americans with chronic renal insufficiency: The African American Study of Kidney Disease and Hypertension Trial. Am. J. Kidney Dis. 2002, 39, 513–524. [Google Scholar] [CrossRef] [PubMed]
- Pagels, A.A.; Soderkvist, B.K.; Medin, C.; Hylander, B.; Heiwe, S. Health-related quality of life in different stages of chronic kidney disease and at initiation of dialysis treatment. Health Qual. Life Outcomes 2012, 10, 71. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Mujais, S.K.; Story, K.; Brouillette, J.; Takano, T.; Soroka, S.; Franek, C.; Mendelssohn, D.; Finkelstein, F.O. Health-related quality of life in CKD Patients: Correlates and evolution over time. Clin. J. Am. Soc. Nephrol. 2009, 4, 1293–1301. [Google Scholar] [CrossRef] [Green Version]
- Perlman, R.L.; Finkelstein, F.O.; Liu, L.; Roys, E.; Kiser, M.; Eisele, G.; Burrows-Hudson, S.; Messana, J.M.; Levin, N.; Rajagopalan, S.; et al. Quality of life in chronic kidney disease (CKD): A cross-sectional analysis in the Renal Research Institute-CKD study. Am. J. Kidney Dis. 2005, 45, 658–666. [Google Scholar] [CrossRef] [PubMed]
- Xie, Y.; Bowe, B.; Mokdad, A.H.; Xian, H.; Yan, Y.; Li, T.; Maddukuri, G.; Tsai, C.Y.; Floyd, T.; Al-Aly, Z. Analysis of the Global Burden of Disease study highlights the global, regional, and national trends of chronic kidney disease epidemiology from 1990 to 2016. Kidney Int. 2018, 94, 567–581. [Google Scholar] [CrossRef] [Green Version]
- Luyckx, V.A.; Tuttle, K.R.; Garcia-Garcia, G.; Gharbi, M.B.; Heerspink, H.J.L.; Johnson, D.W.; Liu, Z.H.; Massy, Z.A.; Moe, O.; Nelson, R.G.; et al. Reducing major risk factors for chronic kidney disease. Kidney Int. Suppl. 2017, 7, 71–87. [Google Scholar] [CrossRef] [Green Version]
- Ameh, O.I.; Ekrikpo, U.; Bello, A.; Okpechi, I. Current Management Strategies of Chronic Kidney Disease in Resource-Limited Countries. Int. J. Nephrol. Renovasc. Dis. 2020, 13, 239–251. [Google Scholar] [CrossRef]
- Stevens, L.A.; Viswanathan, G.; Weiner, D.E. Chronic kidney disease and end-stage renal disease in the elderly population: Current prevalence, future projections, and clinical significance. Adv. Chronic. Kidney Dis. 2010, 17, 293–301. [Google Scholar] [CrossRef] [Green Version]
- Sheng, L.; Zhuang, S. New Insights Into the Role and Mechanism of Partial Epithelial-Mesenchymal Transition in Kidney Fibrosis. Front. Physiol. 2020, 11, 569322. [Google Scholar] [CrossRef] [PubMed]
- Guo, L.; Peng, W.; Tao, J.; Lan, Z.; Hei, H.; Tian, L.; Pan, W.; Wang, L.; Zhang, X. Hydrogen Sulfide Inhibits Transforming Growth Factor-beta1-Induced EMT via Wnt/Catenin Pathway. PLoS ONE 2016, 11, e0147018. [Google Scholar] [CrossRef] [Green Version]
- Lamouille, S.; Xu, J.; Derynck, R. Molecular mechanisms of epithelial-mesenchymal transition. Nat. Rev. Mol. Cell Biol. 2014, 15, 178–196. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Grgic, I.; Duffield, J.S.; Humphreys, B.D. The origin of interstitial myofibroblasts in chronic kidney disease. Pediatr. Nephrol. 2012, 27, 183–193. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Fragiadaki, M.; Mason, R.M. Epithelial-mesenchymal transition in renal fibrosis—Evidence for and against. Int. J. Exp. Pathol. 2011, 92, 143–150. [Google Scholar] [CrossRef] [PubMed]
- Liu, D.; Cheng, F.; Pan, S.; Liu, Z. Stem cells: A potential treatment option for kidney diseases. Stem Cell Res. Ther. 2020, 11, 249. [Google Scholar] [CrossRef]
- Wong, C.Y. Current advances of stem cell-based therapy for kidney diseases. World J. Stem Cells 2021, 13, 914–933. [Google Scholar] [CrossRef]
- Rota, C.; Morigi, M.; Imberti, B. Stem Cell Therapies in Kidney Diseases: Progress and Challenges. Int. J. Mol. Sci. 2019, 20, 2790. [Google Scholar] [CrossRef] [Green Version]
- Chung, B.H. Use of mesenchymal stem cells for chronic kidney disease. Kidney Res. Clin. Pract. 2019, 38, 131–134. [Google Scholar] [CrossRef] [Green Version]
- Yokote, S.; Katsuoka, Y.; Yamada, A.; Ohkido, I.; Yokoo, T. Effect of adipose-derived mesenchymal stem cell transplantation on vascular calcification in rats with adenine-induced kidney disease. Sci. Rep. 2017, 7, 14036. [Google Scholar] [CrossRef] [Green Version]
- Qi, S.; Wu, D. Bone marrow-derived mesenchymal stem cells protect against cisplatin-induced acute kidney injury in rats by inhibiting cell apoptosis. Int. J. Mol. Med. 2013, 32, 1262–1272. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Xiang, E.; Han, B.; Zhang, Q.; Rao, W.; Wang, Z.; Chang, C.; Zhang, Y.; Tu, C.; Li, C.; Wu, D. Human umbilical cord-derived mesenchymal stem cells prevent the progression of early diabetic nephropathy through inhibiting inflammation and fibrosis. Stem Cell Res. Ther. 2020, 11, 336. [Google Scholar] [CrossRef] [PubMed]
- Sun, B.; Luo, X.; Yang, C.; Liu, P.; Yang, Y.; Dong, X.; Yang, Z.; Xu, J.; Zhang, Y.; Li, L. Therapeutic Effects of Human Urine-Derived Stem Cells in a Rat Model of Cisplatin-Induced Acute Kidney Injury In Vivo and In Vitro. Stem Cells Int. 2019, 2019, 8035076. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Zhang, C.; George, S.K.; Wu, R.; Thakker, P.U.; Abolbashari, M.; Kim, T.H.; Ko, I.K.; Zhang, Y.; Sun, Y.; Jackson, J.; et al. Reno-protection of Urine-derived Stem Cells in A Chronic Kidney Disease Rat Model Induced by Renal Ischemia and Nephrotoxicity. Int. J. Biol. Sci. 2020, 16, 435–446. [Google Scholar] [CrossRef] [Green Version]
- Chang, J.W.; Hung, S.P.; Wu, H.H.; Wu, W.M.; Yang, A.H.; Tsai, H.L.; Yang, L.Y.; Lee, O.K. Therapeutic effects of umbilical cord blood-derived mesenchymal stem cell transplantation in experimental lupus nephritis. Cell Transpl. 2011, 20, 245–257. [Google Scholar] [CrossRef]
- Wu, Y.; Zhang, C.; Guo, R.; Wu, D.; Shi, J.; Li, L.; Chu, Y.; Yuan, X.; Gao, J. Mesenchymal Stem Cells: An Overview of Their Potential in Cell-Based Therapy for Diabetic Nephropathy. Stem Cells Int. 2021, 2021, 6620811. [Google Scholar] [CrossRef]
- Liu, Y.; Tang, S.C. Recent Progress in Stem Cell Therapy for Diabetic Nephropathy. Kidney Dis. 2016, 2, 20–27. [Google Scholar] [CrossRef]
- Bharadwaj, S.; Liu, G.; Shi, Y.; Wu, R.; Yang, B.; He, T.; Fan, Y.; Lu, X.; Zhou, X.; Liu, H.; et al. Multipotential differentiation of human urine-derived stem cells: Potential for therapeutic applications in urology. Stem Cells 2013, 31, 1840–1856. [Google Scholar] [CrossRef]
- Tian, S.F.; Jiang, Z.Z.; Liu, Y.M.; Niu, X.; Hu, B.; Guo, S.C.; Wang, N.S.; Wang, Y. Human urine-derived stem cells contribute to the repair of ischemic acute kidney injury in rats. Mol. Med. Rep. 2017, 16, 5541–5548. [Google Scholar] [CrossRef]
- Zhou, Q.; Cheng, Y.; Sun, F.; Shen, J.; Nasser, M.I.; Zhu, P.; Zhang, X.; Li, Y.; Yin, G.; Wang, Y.; et al. A Comprehensive Review of the Therapeutic Value of Urine-Derived Stem Cells. Front. Genet. 2021, 12, 781597. [Google Scholar] [CrossRef]
- Zhang, W.; Hu, J.; Huang, Y.; Wu, C.; Xie, H. Urine-derived stem cells: Applications in skin, bone and articular cartilage repair. Burn. Trauma 2021, 9, tkab039. [Google Scholar] [CrossRef]
- Hu, M.C.; Shi, M.; Zhang, J.; Pastor, J.; Nakatani, T.; Lanske, B.; Razzaque, M.S.; Rosenblatt, K.P.; Baum, M.G.; Kuro-o, M.; et al. Klotho: A novel phosphaturic substance acting as an autocrine enzyme in the renal proximal tubule. FASEB J. 2010, 24, 3438–3450. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Ide, N.; Olauson, H.; Sato, T.; Densmore, M.J.; Wang, H.; Hanai, J.I.; Larsson, T.E.; Lanske, B. In vivo evidence for a limited role of proximal tubular Klotho in renal phosphate handling. Kidney Int. 2016, 90, 348–362. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Sugiura, H.; Yoshida, T.; Shiohira, S.; Kohei, J.; Mitobe, M.; Kurosu, H.; Kuro-o, M.; Nitta, K.; Tsuchiya, K. Reduced Klotho expression level in kidney aggravates renal interstitial fibrosis. Am. J. Physiol. Renal. Physiol. 2012, 302, F1252–F1264. [Google Scholar] [CrossRef] [PubMed]
- Hu, M.C.; Kuro-o, M.; Moe, O.W. Secreted klotho and chronic kidney disease. Adv. Exp. Med. Biol. 2012, 728, 126–157. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Doi, S.; Zou, Y.; Togao, O.; Pastor, J.V.; John, G.B.; Wang, L.; Shiizaki, K.; Gotschall, R.; Schiavi, S.; Yorioka, N.; et al. Klotho inhibits transforming growth factor-beta1 (TGF-beta1) signaling and suppresses renal fibrosis and cancer metastasis in mice. J. Biol. Chem. 2011, 286, 8655–8665. [Google Scholar] [CrossRef] [Green Version]
- Hu, M.C.; Shi, M.; Zhang, J.; Quinones, H.; Griffith, C.; Kuro-o, M.; Moe, O.W. Klotho deficiency causes vascular calcification in chronic kidney disease. J. Am. Soc. Nephrol. 2011, 22, 124–136. [Google Scholar] [CrossRef]
- Shimamura, Y.; Hamada, K.; Inoue, K.; Ogata, K.; Ishihara, M.; Kagawa, T.; Inoue, M.; Fujimoto, S.; Ikebe, M.; Yuasa, K.; et al. Serum levels of soluble secreted alpha-Klotho are decreased in the early stages of chronic kidney disease, making it a probable novel biomarker for early diagnosis. Clin. Exp. Nephrol. 2012, 16, 722–729. [Google Scholar] [CrossRef]
- Koh, N.; Fujimori, T.; Nishiguchi, S.; Tamori, A.; Shiomi, S.; Nakatani, T.; Sugimura, K.; Kishimoto, T.; Kinoshita, S.; Kuroki, T.; et al. Severely reduced production of klotho in human chronic renal failure kidney. Biochem. Biophys. Res. Commun. 2001, 280, 1015–1020. [Google Scholar] [CrossRef]
- Chen, L.; Li, L.; Xing, F.; Peng, J.; Peng, K.; Wang, Y.; Xiang, Z. Human Urine-Derived Stem Cells: Potential for Cell-Based Therapy of Cartilage Defects. Stem Cells Int. 2018, 2018, 4686259. [Google Scholar] [CrossRef] [Green Version]
- Luo, S.; Shao, L.; Geng, R.; Liu, Q.; Jiang, W.; Gong, M.; Zhang, Y.; He, Y. Identification and biological characteristics of clear cell renal cell carcinoma associated urine-derived stem cells. Am. J. Transl. Res. 2021, 13, 2143–2162. [Google Scholar] [PubMed]
- Sun, J.; Xing, F.; Zou, M.; Gong, M.; Li, L.; Xiang, Z. Comparison of chondrogenesis-related biological behaviors between human urine-derived stem cells and human bone marrow mesenchymal stem cells from the same individual. Stem Cell Res. Ther. 2021, 12, 366. [Google Scholar] [CrossRef] [PubMed]
- Azuma, M.; Koyama, D.; Kikuchi, J.; Yoshizawa, H.; Thasinas, D.; Shiizaki, K.; Kuro-o, M.; Furukawa, Y.; Kusano, E. Promoter methylation confers kidney-specific expression of the Klotho gene. FASEB J. 2012, 26, 4264–4274. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Yan, J.D.; Yang, S.; Zhang, J.; Zhu, T.H. BMP6 reverses TGF-beta1-induced changes in HK-2 cells: Implications for the treatment of renal fibrosis. Acta Pharmacol. Sin. 2009, 30, 994–1000. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Tian, Y.C.; Phillips, A.O. TGF-beta1-mediated inhibition of HK-2 cell migration. J. Am. Soc. Nephrol. 2003, 14, 631–640. [Google Scholar] [CrossRef] [Green Version]
- Lawson, J.S.; Liu, H.H.; Syme, H.M.; Purcell, R.; Wheeler-Jones, C.P.D.; Elliott, J. The cat as a naturally occurring model of renal interstitial fibrosis: Characterisation of primary feline proximal tubular epithelial cells and comparative pro-fibrotic effects of TGF-beta1. PLoS ONE 2018, 13, e0202577. [Google Scholar] [CrossRef]
- Meng, X.M.; Tang, P.M.; Li, J.; Lan, H.Y. TGF-beta/Smad signaling in renal fibrosis. Front. Physiol. 2015, 6, 82. [Google Scholar] [CrossRef] [Green Version]
- Zhang, Y.E. Non-Smad pathways in TGF-beta signaling. Cell Res. 2009, 19, 128–139. [Google Scholar] [CrossRef] [Green Version]
- Benigni, A.; Morigi, M.; Remuzzi, G. Kidney regeneration. Lancet 2010, 375, 1310–1317. [Google Scholar] [CrossRef] [Green Version]
- Menn-Josephy, H.; Lee, C.S.; Nolin, A.; Christov, M.; Rybin, D.V.; Weinberg, J.M.; Henderson, J.; Bonegio, R.; Havasi, A. Renal Interstitial Fibrosis: An Imperfect Predictor of Kidney Disease Progression in Some Patient Cohorts. Am. J. Nephrol. 2016, 44, 289–299. [Google Scholar] [CrossRef] [Green Version]
- Qin, D.; Long, T.; Deng, J.; Zhang, Y. Urine-derived stem cells for potential use in bladder repair. Stem Cell Res. Ther. 2014, 5, 69. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Buchanan, S.; Combet, E.; Stenvinkel, P.; Shiels, P.G. Klotho, Aging, and the Failing Kidney. Front. Endocrinol. 2020, 11, 560. [Google Scholar] [CrossRef] [PubMed]
- Eltablawy, N.; Ashour, H.; Rashed, L.A.; Hamza, W.M. Vitamin D protection from rat diabetic nephropathy is partly mediated through Klotho expression and renin-angiotensin inhibition. Arch. Physiol. Biochem. 2018, 124, 461–467. [Google Scholar] [CrossRef] [PubMed]
- Drew, D.A.; Katz, R.; Kritchevsky, S.; Ix, J.; Shlipak, M.; Gutierrez, O.M.; Newman, A.; Hoofnagle, A.; Fried, L.; Semba, R.D.; et al. Association between Soluble Klotho and Change in Kidney Function: The Health Aging and Body Composition Study. J. Am. Soc. Nephrol. 2017, 28, 1859–1866. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Kuro, O.M. Phosphate and Klotho. Kidney Int. Suppl. 2011, 79, S20–S23. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Sanchez-Nino, M.D.; Fernandez-Fernandez, B.; Ortiz, A. Klotho, the elusive kidney-derived anti-ageing factor. Clin. Kidney J. 2020, 13, 125–127. [Google Scholar] [CrossRef] [PubMed]
- Zhou, L.; Li, Y.; Zhou, D.; Tan, R.J.; Liu, Y. Loss of Klotho contributes to kidney injury by derepression of Wnt/beta-catenin signaling. J. Am. Soc. Nephrol. 2013, 24, 771–785. [Google Scholar] [CrossRef] [Green Version]
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Kim, S.-H.; Jin, J.-A.; So, H.J.; Lee, S.H.; Kang, T.-W.; Lee, J.-U.; Choi, D.E.; Jeong, J.Y.; Chang, Y.-K.; Choi, H.; et al. Urine-Derived Stem Cell-Secreted Klotho Plays a Crucial Role in the HK-2 Fibrosis Model by Inhibiting the TGF-β Signaling Pathway. Int. J. Mol. Sci. 2022, 23, 5012. https://doi.org/10.3390/ijms23095012
Kim S-H, Jin J-A, So HJ, Lee SH, Kang T-W, Lee J-U, Choi DE, Jeong JY, Chang Y-K, Choi H, et al. Urine-Derived Stem Cell-Secreted Klotho Plays a Crucial Role in the HK-2 Fibrosis Model by Inhibiting the TGF-β Signaling Pathway. International Journal of Molecular Sciences. 2022; 23(9):5012. https://doi.org/10.3390/ijms23095012
Chicago/Turabian StyleKim, Sang-Heon, Jeong-Ah Jin, Hyung Joon So, Sung Hoon Lee, Tae-Wook Kang, Jae-Ung Lee, Dae Eun Choi, Jin Young Jeong, Yoon-Kyung Chang, Hyunsu Choi, and et al. 2022. "Urine-Derived Stem Cell-Secreted Klotho Plays a Crucial Role in the HK-2 Fibrosis Model by Inhibiting the TGF-β Signaling Pathway" International Journal of Molecular Sciences 23, no. 9: 5012. https://doi.org/10.3390/ijms23095012