DOI QR코드

DOI QR Code

SIRT1 Knockdown Enhances the Differentiation of Human Embryonic Stem Cells into Pancreatic β Cells

  • Seo, Nan-Hee (Dept. of Microbiology, Jeonbuk National University Medical School) ;
  • Song, Hwa-Ryung (Dept. of Microbiology, Jeonbuk National University Medical School) ;
  • Han, Myung-Kwan (Dept. of Microbiology, Jeonbuk National University Medical School)
  • 투고 : 2019.10.21
  • 심사 : 2019.11.25
  • 발행 : 2019.12.31

초록

Nicotinamide is used to maturate pancreatic progenitors from embryonic stem cells (ESCs) into insulin-producing cells (IPCs). It has been known that nicotinamide inhibits the enzymatic activity of SIRT1, an NAD+-dependent deacetylase. Here we show that SIRT1 knockdown enhances the differentiation of human ESCs into IPCs. SIRT1 knockdown enhances the clustering size of IPCs and the expression of pancreatic genes including c-peptide, pancreas/duodenum homeobox protein 1 (PDX1), insulin, somatostatin, glucagon and Nkx6.1 in human ESC-derived IPCs. In addition, We found that IPCs differentiated from SIRT1 knockdowned human ESCs have more zinc compared to those from control human ESCs. Our data suggest that SIRT1 negatively regulates the differentiation of β cells from human ESCs.

키워드

참고문헌

  1. Avalos JL, Bever KM, Wolberger C (2005) Mechanism of sirtuin inhibition by nicotinamide: Altering the NAD(+) cosubstrate specificity of a Sir2 enzyme. Mol Cell 17:855-868. https://doi.org/10.1016/j.molcel.2005.02.022
  2. Baur JA, Chen D, Chini EN, Chua K, Cohen HY, de Cabo R, Deng C, Dimmeler S, Gius D, Guarente LP, Helfand SL, Imai S, Itoh H, Kadowaki T, Koya D, Leeuwenburgh C, McBurney M, Nabeshima Y, Neri C, Oberdoerffer P, Pestell RG, Rogina B, Sadoshima J, Sartorelli V, Serrano M, Sinclair DA, Steegborn C, Tatar M, Tissenbaum HA, Tong Q, Tsubota K, Vaquero A, Verdin E (2010) Dietary restriction: Standing up for sirtuins. Science 329:1012-1013. https://doi.org/10.1126/science.329.5995.1012
  3. Bellin MD, Barton FB, Heitman A, Harmon JV, Kandaswamy R, Balamurugan AN, Sutherland DE, Alejandro R, Hering BJ (2012) Potent induction immunotherapy promotes long-term insulin independence after islet transplantation in type 1 diabetes. Am J Transplant 12:1576-1583. https://doi.org/10.1111/j.1600-6143.2011.03977.x
  4. Bordone L, Motta MC, Picard F, Robinson A, Jhala US, Apfeld J, Mcdonagh T, Lemieux M, Mcburney M, Szilvasi A, Easlon EJ, Lin SJ, Guarente L (2006) Sirt1 regulates insulin secretion by repressing UCP2 in pancreatic beta cells. PLOS Biol 4:e31. https://doi.org/10.1371/journal.pbio.0040031
  5. Donmez G, Guarente L (2010) Aging and disease: Connections to sirtuins. Aging Cell 9:285-290. https://doi.org/10.1111/j.1474-9726.2010.00548.x
  6. Doss MX, Koehler CI, Gissel C, Hescheler J, Sachinidis A (2004) Embryonic stem cells: A promising tool for cell replacement therapy. J Cell Mol Med 8:465-473. https://doi.org/10.1111/j.1582-4934.2004.tb00471.x
  7. Haigis MC, Sinclair DA (2010) Mammalian sirtuins: Biological insights and disease relevance. Annu Rev Pathol 5:253-295. https://doi.org/10.1146/annurev.pathol.4.110807.092250
  8. Han MK, Song EK, Guo Y, Ou X, Mantel C, Broxmeyer HE (2008) SIRT1 regulates apoptosis and Nanog expression in mouse embryonic stem cells by controlling p53 subcellular localization. Cell Stem Cell 2:241-251. https://doi.org/10.1016/j.stem.2008.01.002
  9. Kroon E, Martinson LA, Kadoya K, Bang AG, Kelly OG, Eliazer S, Young H, Richardson M, Smart NG, Cunningham J, Agulnick AD, D'Amour KA, Carpenter MK, Baetge EE (2008) Pancreatic endoderm derived from human embryonic stem cells generates glucose-responsive insulin-secreting cells in vivo. Nat Biotechnol 26:443-452. https://doi.org/10.1038/nbt1393
  10. Kume S, Haneda M, Kanasaki K, Sugimoto T, Araki S, Isono M, Isshiki K, Uzu T, Kashiwagi A, Koya D (2006) Silent information regulator 2 (SIRT1) attenuates oxidative stress-induced mesangial cell apoptosis via p53 deacetylation. Free Radic Biol Med 40:2175-2182. https://doi.org/10.1016/j.freeradbiomed.2006.02.014
  11. Latif ZA, Noel J, Alejandro R (1988) A simple method of staining fresh and cultured islets. Transplantation 45:827- 830. https://doi.org/10.1097/00007890-198804000-00038
  12. Lumelsky N, Blondel O, Laeng P, Velasco I, Ravin R, Mckay R (2001) Differentiation of embryonic stem cells to insulin-secreting structures similar to pancreatic islets. Science 292:1389-1394. https://doi.org/10.1126/science.1058866
  13. Mao GH, Chen GA, Bai HY, Song TR, Wang YX (2009) The reversal of hyperglycaemia in diabetic mice using PLGA scaffolds seeded with islet-like cells derived from human embryonic stem cells. Biomaterials 30:1706-1714. https://doi.org/10.1016/j.biomaterials.2008.12.030
  14. Nishikawa S, Jakt LM, Era T (2007) Embryonic stem-cell culture as a tool for developmental cell biology. Nat Rev Mol Cell Biol 8:502-507. https://doi.org/10.1038/nrm2189
  15. Rezania A, Bruin JE, Riedel MJ, Mojibian M, Asadi A, Xu J, Gauvin R, Narayan K, Karanu F, O'neil JJ, Ao Z, Warnock GL, Kieffer TJ (2012) Maturation of human embryonic stem cell-derived pancreatic progenitors into functional islets capable of treating pre-existing diabetes in mice. Diabetes 61:2016-2029. https://doi.org/10.2337/db11-1711
  16. Sander M, Sussel L, Conners J, Scheel D, Kalamaras J, Dela Cruz F, Schwitzgebel V, Hayes-Jordan A, German M (2000) Homeobox gene Nkx6.1 lies downstream of Nkx2.2 in the major pathway of beta-cell formation in the pancreas. Development 127:5533-5540. https://doi.org/10.1242/dev.127.24.5533
  17. Schisler JC, Fueger PT, Babu DA, Hohmeier HE, Tessem JS, Lu D, Becker TC, Naziruddin B, Levy M, Mirmira RG, Newgard CB (2008) Stimulation of human and rat islet beta-cell proliferation with retention of function by the homeodomain transcription factor Nkx6.1. Mol Cell Biol 28:3465-3476. https://doi.org/10.1128/MCB.01791-07
  18. Steiner DF, Cunningham D, Spigelman L, Aten B (1967) Insulin biosynthesis: Evidence for a precursor. Science 157:697-700. https://doi.org/10.1126/science.157.3789.697
  19. Takahashi K, Yamanaka S (2006) Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell 126:663-676. https://doi.org/10.1016/j.cell.2006.07.024
  20. Thomson JA, Itskovitz-Eldor J, Shapiro SS, Waknitz MA, Swiergiel JJ, Marshall VS, Jones JM (1998) Embryonic stem cell lines derived from human blastocysts. Science 282:1145-1147. https://doi.org/10.1126/science.282.5391.1145
  21. Vaziri H, Dessain SK, Ng Eaton E, Imai SI, Frye RA, Pandita TK, Guarente L, Weinberg RA (2001) hSIR2 (SIRT1) functions as an NAD-dependent p53 deacetylase. Cell 107:149-159. https://doi.org/10.1016/S0092-8674(01)00527-X
  22. Wei R, Yang J, Hou W, Liu G, Gao M, Zhang L, Wang H, Mao G, Gao H, Chen G, Hong T. (2013) Insulin-producing cells derived from human embryonic stem cells: Comparison of definitive endoderm- and nestin-positive progenitor-based differentiation strategies. PLOS ONE 8:e72513. https://doi.org/10.1371/journal.pone.0072513