Histone Deacetylase Inhibitor (Trapoxin A) Enhances Stemness Properties in Adipose Tissue Derived Mesenchymal Stem Cells

Abstract

Back ground Adipose tissue derived mesenchymal stem cells (ASCs) have unique potential for regenerative cell therapies. However, during ex-vivo cultivation, they undergo considerable quality loss regarding their phenotypic properties, stemness genes expression and differentiation potential. Recent studies reported that the loss of stemness properties of MSCs is a result of chromatin histone deacetylations through in-vitro cultivation. The present work aimed to study the effect of Trapoxin A (TPX) as a histone deacetylase inhibitor (HDACi) on overall stemness properties of ASCs.

Methods First, the effects of TPX treatments on ASCs viability and proliferation were evaluated using MTT assay. Second, the desired doses of TPX supporting ASCs proliferation were determined and the lack of their negative effects was confirmed by DAPI staining. In addition, the influence of TPX on cell cycle of ASCs and the mRNA levels of stemness genes were measured by flowcytometry and qPCR, respectively. Finally, the effect of TPX treatment on osteogenic potential of ASCs was studied.

Results The results indicated that short time TPX treatment (nM concentrations) caused stimulation of proliferation and considerable percentage of ASCs entered to S-phase of cell cycle (p<0.05). Moreover, the findings demonstrated significant up-regulation of stemness markers genes (Oct-4, Sox-2, Nanog, TERT, Klf-4, Rex-1) (p<0.05) and enhanced osteogenic differentiation potential of ASC after TPX treatment.

Conclusion The addition of low dose of TPX to the expansion medium could possibly enhance the stemness properties and prevent the quality decline of ex-vivo cultured ASCs.

Key words

Adipose tissue mesenchymal stem cells - Stemness - Trapoxin A

• References

• 1 Alizadeh E, Akbarzadeh A, Eslaminejad MB. et al Up Regulation of Liver-enriched Transcription Factors HNF4a and HNF6 and Liver-Specific MicroRNA (miR-122) by Inhibition of Let-7b in Mesenchymal Stem Cells. Chemical biology & drug design 2015; 85: 268-279
• 2 Alizadeh E, Zarghami N, Eslaminejad MB. et al. The effect of dimethyl sulfoxide on hepatic differentiation of mesenchymal stem cells. Artificial cells, nanomedicine, and biotechnology 2016; 44: 157-164
• 3 Zhang J, Huang X, Wang H. et al. The challenges and promises of allogeneic mesenchymal stem cells for use as a cell-based therapy. Stem Cell Research & Therapy 2015; 6: 234
• 4 Turgeman G. The therapeutic potential of mesenchymal stem cells in Alzheimer's disease: converging mechanisms. Neural Regeneration Research 2015; 10: 698-699
• 5 Monfrini M, Donzelli E, Rodriguez-Menendez V. et al. Therapeutic potential of mesenchymal stem cells for the treatment of diabetic peripheral neuropathy. Experimental neurology 2017; 288: 75-84
• 6 Laube M, Stolzing A, Thome UH. et al. Therapeutic potential of mesenchymal stem cells for pulmonary complications associated with preterm birth. The International Journal of Biochemistry & Cell Biology 2016; 74: 18-32
• 7 Saei Arezoumand K, Alizadeh E, Pilehvar-Soltanahmadi Y et al. An overview on different strategies for the stemness maintenance of MSCs. Artificial Cells Nano medicine & Biotechnology 2016
• 8 Li Z, Liu C, Xie Z. et al. Epigenetic dysregulation in mesenchymal stem cell aging and spontaneous differentiation. PloS one 2011; 6: e20526
• 9
• 10 Li Y, Chu JS, Kurpinski K. et al. Biophysical regulation of histone acetylation in mesenchymal stem cells. Biophysical Journal 2011; 100: 1902-1909
• 11 Sadeghi Z, Nejati-koshki K, Akbarzadeh A. et al. Trichostatin A-induced apoptosis is mediated by krüppel-like factor 4 in ovarian and lung cancer. Asian Pacific Journal of Cancer Prevention 2014; 15: 6581-6586
• 12 Karantzali E, Schulz H, Hummel O. et al. Histone deacetylase inhibition accelerates the early events of stem cell differentiation: Transcriptomic and epigenetic analysis. Genome Biol 2008; 9: R65
• 13 Kijima M, Yoshida M, Sugita K. et al. Trapoxin, an antitumor cyclic tetrapeptide, is an irreversible inhibitor of mammalian histone deacetylase. The Journal of biological chemistry 1993; 268: 22429-22435
• 14 Yoshida M, Horinouchi S, Beppu T. et al. and trapoxin: novel chemical probes for the role of histone acetylation in chromatin structure and function. Bioessays 1995; 17: 423-430
• 15 Alizadeh E, Eslaminejad MB, Akbarzadeh A. et al. Upregulation of MiR-122 via trichostatin a treatments in hepatocyte-like cells derived from mesenchymal stem cells. Chemical biology & drug design 2016; 87: 296-305
• 16 Hoseinzadeh S, Atashi A, Soleimani M. et al. MiR-221-inhibited adipose tissue-derived mesenchymal stem cells bioengineered in a nano-hydroxy apatite scaffold. In Vitro Cellular & Developmental Biology-Animal 2016; 52: 479-487
• 17 Majidinia M, Alizadeh E, Yousefi B. et al. Co-inhibition of notch and nf-κb signaling pathway decreases proliferation through downregulating iκb-α and hes-1 expression in human ovarian cancer OVCAR-3 cells. Drug research 2017; 67: 13-19
• 18 Wang D-Z, Gao J-F, Jing S-F. et al. Antitumor effect of docetaxel in osteosarcoma by the inhibition of Wnt signal channel. Drug research 2015; 65: 597-601
• 19 Mousazadeh H, Milani M, Zarghami N et al. Study of the cytotoxic and bactericidal effects of sila-substituted thioalkyne and mercaptothionecompounds based on 1, 2, 3-Triazole Scaffold. Basic & Clinical Pharmacology & Toxicology 2017
• 20 Saki F, Sheikhi A, Omrani GHR. et al. Evaluation of bone mineral density in children with type i diabetes mellitus and relationship to serum levels of osteopontin. Drug research 2017; 67: 527-533
• 21 Shotorbani BB, Alizadeh E, Salehi R. et al. Adhesion of mesenchymal stem cells to biomimetic polymers: A review. Materials Science and Engineering: C 2017; 71: 1192-1200
• 22 Bruder SP, Jaiswal N, Haynesworth SE. Growth kinetics, self-renewal, and the osteogenic potential of purified human mesenchymal stem cells during extensive subcultivation and following cryopreservation. Journal of cellular biochemistry 1997; 64: 278-294
• 23 Wagner W, Horn P, Castoldi M. et al. Replicative senescence of mesenchymal stem cells: a continuous and organized process. PloS one 2008; 3: e2213
• 24 Kapetanou M, Chondrogianni N, Petrakis S. et al. Proteasome activation enhances stemness and lifespan of human mesenchymal stem cells. Free Radical Biology and Medicine 2017; 103: 226-235
• 25 Agalioti T, Chen G, Thanos D. Deciphering the transcriptional histone acetylation code for a human gene. Cell 2002; 111: 381-392
• 26 Lomvardas S, Thanos D. Nucleosome sliding via TBP DNA binding in vivo. Cell 2001; 106: 685-696
• 27 Richon VM, Sandhoff TW, Rifkind RA. et al. Histone deacetylase inhibitor selectively induces p21WAF1 expression and gene-associated histone acetylation. Proceedings of the National Academy of Sciences 2000; 97: 10014-10019
• 28 Han B, Li J, Li Z. et al. Stabilizes the Expression of Pluripotent Genes in Human Mesenchymal Stem Cells during Ex Vivo Expansion. PLoS ONE 2013; 8: e81781
• 29 Young J, Wu S, Hansteen G. et al. Inhibitors of histone deacetylases promote hematopoietic stem cell self-renewal. Cytotherapy 2004; 6: 328-336
• 30 Hu X, Zhang X, Dai L. et al. Histone deacetylase inhibitor trichostatin A promotes the osteogenic differentiation of rat adipose-derived stem cells by altering the epigenetic modifications on Runx2 promoter in a BMP signaling-dependent manner. Stem cells and development 2013; 22: 248-255
• 31 Tonti GA, Mannello F. From bone marrow to therapeutic applications: Different behaviour and genetic/epigenetic stability during mesenchymal stem cell expansion in autologous and foetal bovine sera?. International Journal of Developmental Biology 2002; 52: 1023-1032