The Histone Deacetylase Inhibitor Sodium Butyrate Stimulates Adipogenesis Through a Limited Transcriptional Switch in Periodontal Ligament-Derived Stem Cells
Keywords:
Periodontal ligament stem cells; Histone acetylation; Adipogenesis; Histone deacetylase inhibitors.Abstract
Adipogenic differentiation plays a crucial role in adipose tissue biology, an endocrine organ that regulates energy storage and hormone secretion. Dysfunction in this process contributes to metabolic diseases such as obesity and type II diabetes. In vitro models have been developed to investigate the mechanisms of adipogenesis, with periodontal ligament stem cells (PDLSCs) emerging as a promising model due to their multipotent capacity. Previous studies have shown that epigenetic manipulation can enhance the adipogenic response in various cell lines. Acetylation of lysine 9 on histone H3 (H3K9ac) is associated with the activation of key genes, such as PPARγ-2. In this study, we evaluated whether class I histone deacetylase inhibitors, such as valproic acid (VPA) and sodium butyrate (NaBu), both short-chain fatty acids, can increase H3K9 acetylation and influence adipogenic differentiation. We used 1, 4, and 8 mM VPA concentrations and 1, 2, and 5 mM NaBu to assess their effects on cell viability, morphology, H3K9ac distribution, and adipogenic differentiation. The results indicated that cells treated with 5 mM NaBu exhibited morphological changes, reduced viability, increased H3K9ac signal intensity, and enhanced intracellular lipid deposition. These results infer that inhibition of HDACs by NaBu increases plasticity toward adipogenesis of PDLSCs through a limited transcriptional change in their key genes.
References
De Fano M., Malara M., Vermigli C., Murdolo G. Adipose Tissue: A Novel Target of the Incretin Axis? A Paradigm Shift in Obesity-Linked Insulin Resistance. Int J Mol Sci. 2024; 25 (16).
Rodríguez-Fuentes D.E., Fernández-Garza L.E., Samia-Meza J.A., Barrera-Barrera S.A., Caplan A.I., Barrera-Saldaña H.A. Mesenchymal Stem Cells Current Clinical Applications: A Systematic Review. Arch Med Res. 2021; 52 (1): 93-101.
Thant L., Kaku M., Kakihara Y., Mizukoshi M., Kitami M., Arai M., et al. Extracellular Matrix-Oriented Proteomic Analysis of Periodontal Ligament Under Mechanical Stress. Front Physiol. 2022; 13: 899699.
Iwayama T., Sakashita H., Takedachi M., Murakami S. Periodontal tissue stem cells and mesenchymal stem cells in the periodontal ligament. Jpn Dent Sci Rev. 2022; 58: 172-8.
Trejo Iriarte C.G., Ramírez Ramírez O., Muñoz García A., Verdín Terán S.L., Gómez Clavel J.F. Aislamiento de células mesenquimales del ligamento periodontal de premolares extraídos. Método simplificado. Rev. Odont. Mex. 2017; 21 (1): 13-21.
Wu Y., Wang Y., Ji Y., Ou Y., Xia H., Zhang B., et al. C4orf7 modulates osteogenesis and adipogenesis of human periodontal ligament cells. Am J Transl Res. 2017; 9 (12): 5708.
Matsushita K., Dzau V.J. Mesenchymal stem cells in obesity: insights for translational applications. Lab. Invest. 2017; 97 (10): 1158-66.
Musri M.M., Gomis R., Párrizas M. A chromatin perspective of adipogenesis. Organogenesis. 2010; 6 (1): 15-23.
Argaez-Sosa A.A., Rodas-Junco B.A., Carrillo-Cocom L.M., Rojas-Herrera R.A., Coral-Sosa A., Aguilar-Ayala F.J., et al. Higher Expression of DNA (de)methylation-Related Genes Reduces Adipogenicity in Dental Pulp Stem Cells. Front Cell Dev Biol. 2022; 10: 791667.
Musri M.M., Gomis R., Párrizas M. Chromatin and chromatin-modifying proteins in adipogenesis. Biochem Cell Biol. 2007; 85 (4): 397-410.
Zhang Q., Ramlee M.K., Brunmeir R., Villanueva C.J., Halperin D., Xu F. Dynamic and distinct histone modifications modulate the expression of key adipogenesis regulatory genes. Cell Cycle. 2012; 11 (23): 4310-22.
Hu W., Jiang C., Kim M., Xiao Y., Richter H.J., Guan D., et al. Isoform-specific functions of PPARγ in gene regulation and metabolism. Genes Dev. 2022; 36 (5-6): 300-12.
Fu Y., Luo N., Klein R.L., Garvey W.T. Adiponectin promotes adipocyte differentiation, insulin sensitivity, and lipid accumulation. J Lipid Res. 2005; 46 (7): 1369-79.
Mortada I., Mortada R. Epigenetic changes in mesenchymal stem cells differentiation. Eur J Med Genet. 2018; 61 (2): 114-8.
Steger DJ, Grant GR, Schupp M, Tomaru T, Lefterova MI, Schug J, et al. Propagation of adipogenic signals through an epigenomic transition state. Genes Dev. 2010; 24 (10): 1035-44.
King J., Patel M., Chandrasekaran S. Metabolism, HDACs, and HDAC Inhibitors: A Systems Biology Perspective. Metabolites. 2021; 11 (1): 792.
Jang S., Hwang J., Jeong H-S. The Role of Histone Acetylation in Mesenchymal Stem Cell Differentiation. Chonnam Med J. 2022; 58 (1): 6-12.
Yoo E.J., Chung J.J., Choe S.S., Kim K.H., Kim J.B. Down-regulation of histone deacetylases stimulates adipocyte differentiation. J Biol Chem. 2006; 281 (10): 6608-15.
Kuzmochka C., Abdou H-S, Haché R.J.G., Atlas E. Inactivation of Histone Deacetylase 1 (HDAC1) But Not HDAC2 Is Required for the Glucocorticoid-Dependent CCAAT/Enhancer-Binding Protein α (C/EBPα) Expression and Preadipocyte Differentiation. Endocrinology. 2014; 155 (12): 4762-73.
Pant R., Alam A., Choksi A., Shah V.K., Firmal P., Chattopadhyay S. Chromatin remodeling protein SMAR1 regulates adipogenesis by modulating the expression of PPARγ. Biochim. Biophys. Acta Mol. Cell Biol. Lipids. 2021; 1866 (12): 159045.
Lagace D.C., Nachtigal M.W. Inhibition of histone deacetylase activity by valproic acid blocks adipogenesis. J Biol Chem. 2004; 279 (18): 18851-60.
Ho R.H., Chan J.C.Y., Fan H., Kioh D.Y.Q., Lee B.W., Chan E.C.Y. In Silico and in Vitro Interactions between Short Chain Fatty Acids and Human Histone Deacetylases. Biochemistry. 2017; 56 (36): 4871-8.
Sixto-López Y., Bello M., Correa-Basurto J. Exploring the inhibitory activity of valproic acid against the HDAC family using an MMGBSA approach. J Comput Aided Mol Des. 2020; 34 (8): 857-78.
Romoli M., Mazzocchetti P., D’Alonzo R., Siliquini S., Rinaldi V.E., Verrotti A., et al. Valproic Acid and Epilepsy: From Molecular Mechanisms to Clinical Evidences. Curr Neuropharmacol. 2019; 17 (10): 926-46.
Fu Y., Zhang P., Ge J., Cheng J., Dong W., Yuan H., et al. Histone deacetylase 8 suppresses osteogenic differentiation of bone marrow stromal cells by inhibiting histone H3K9 acetylation and RUNX2 activity. Int J Biochem Cell Biol. 2014; 54: 68-77.
Santos J., Hubert T., Milthorpe B.K. Valproic Acid Promotes Early Neural Differentiation in Adult Mesenchymal Stem Cells Through Protein Signalling Pathways. Cells. 2020; 9 (3).
Um S., Lee H., Zhang Q., Kim H.Y., Lee J-H, Seo B.M. Valproic Acid Modulates the Multipotency in Periodontal Ligament Stem Cells via p53-Mediated Cell Cycle. Tissue Eng Regen Med. 2017; 14 (2):153-62.
Serralta-Interian A., Toro J., Nic Can G., Rojas Herrera R., Aguilar-Ayala F.J., Rodas-Junco B. Inhibition of histone deacetylases class I improves adipogenic differentiation of human periodontal ligament cells. Cell Mol Biol. 2024; 70: 40-7.
Rashid S., Salim A., Qazi R-E-M, Malick T.S., Haneef K. Sodium Butyrate Induces Hepatic Differentiation of Mesenchymal Stem Cells in 3D Collagen Scaffolds. Appl Biochem Biotechnol. 2022; 194 (8): 3721-32.
Eung J.Y., Chung J.J., Sung S.C., Kang H.K., Jae B.K. Down-regulation of histone deacetylases stimulates adipocyte differentiation. J Biol chem. 2006; 281 (10): 6608-15.
Lee H., Lee J.Y., Ha D-H, Jeong J-H, Park J-B. Effects of Valproic Acid on Morphology, Proliferation, and Differentiation of Mesenchymal Stem Cells Derived From Human Gingival Tissue. Implant Dent. 2018; 27 (1): 33-42.
Jones J., Juengel E., Mickuckyte A., Hudak L., Wedel S., Jonas D., et al. Valproic acid blocks adhesion of renal cell carcinoma cells to endothelium and extracellular matrix. J Cell Mol. 2009; 13 (8b): 2342.
Knox E.G., Aburto M.R., Tessier C., Nagpal J., Clarke G., O’Driscoll C.M., et al. Microbial-derived metabolites induce actin cytoskeletal rearrangement and protect blood-brain barrier function. iScience. 2022; 25 (12): 105648.
Fock E., Parnova R. Mechanisms of Blood-Brain Barrier Protection by Microbiota-Derived Short-Chain Fatty Acids. Cells. 2023; 12 (4).
López-García J., Lehocký M., Humpolíček P., Sáha P. HaCaT Keratinocytes Response on Antimicrobial Atelocollagen Substrates: Extent of Cytotoxicity, Cell Viability and Proliferation. J Funct Biomater. 2014; 5 (2): 43-57.
Yu Y., Oh S-Y, Kim H.Y., Choi J-Y, Jo S.A., Jo I. Valproic Acid-Induced CCN1 Promotes Osteogenic Differentiation by Increasing CCN1 Protein Stability through HDAC1 Inhibition in Tonsil-Derived Mesenchymal Stem Cells. Cells. 2022; 11 (3).
Ma X-J, Wang Y-S, Gu W-P, Zhao X. The role and possible molecular mechanism of valproic acid in the growth of MCF-7 breast cancer cells. Croat Med J. 2017; 58 (5): 349-57.
Dowker-Key P.D., Jadi P.K., Gill N.B., Hubbard K.N., Elshaarrawi A., Alfatlawy N.D., et al. A Closer Look into White Adipose Tissue Biology and the Molecular Regulation of Stem Cell Commitment and Differentiation. Genes. 2024; 15 (8): 1017.
Nunn A.D.G., Scopigno T., Pediconi N., Levrero M., Hagman H., Kiskis J., et al. The histone deacetylase inhibiting drug Entinostat induces lipid accumulation in differentiated HepaRG cells. Sci Rep. 2016; 6: 37204.
Tugnoli B., Bernardini C., Forni M., Piva A., Stahl C.H., Grilli E. Butyric acid induces spontaneous adipocytic differentiation of porcine bone marrow-derived mesenchymal stem cells. In Vitro Cell Dev Biol Anim. 2019; 55 (1): 17-24.
Stachecka J., Kolodziejski P.A., Noak M., Szczerbal I. Alteration of active and repressive histone marks during adipogenic differentiation of porcine mesenchymal stem cells. Sci Rep. 2021; 11 (1): 1325.
Montero-Del-Toro J.A., Serralta-Interian A.A., Nic-Can G.I., Rojas-Herrera R., Carrillo-Cocom L.M., Rodas-Junco B.A. Effect of Epigenetic Inhibitors on Adipogenesis in Human Periodontal Ligament Stem Cells. Odovtos-Int J Dent Sc. 2024; 116-128.
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