Abstract
Adipose derived stem cells (ADSCs) have been increasingly explored for use in cell-based therapy against ischemic diseases. However, unsatisfactory angiogenesis limits the therapeutic efficacy. Netrin-1, a known axon guidance molecule, improves neovascularization in the ischemic region. Thus, our study was performed to evaluate the potential effect of Netrin-1 on the angiogenic behaviors of human ADSCs (hADSCs). hADSCs acquired from human abdominal adipose tissue were modified by liposome transfection of Netrin-1 plasmid, and the proliferation of hADSCs was determined by Cell Counting Kit-8 (CCK-8) assay. The transcript levels of pro-invasive proteins such as matrix metalloproteinase 2 (MMP2) and matrix metalloproteinase 9 (MMP-9), were measured to test migratory and invasive capabilities, and the levels of vascular endothelial growth factors were assayed to monitor angiogenic activity. Our results showed that Netrin-1 overexpression enhanced the proliferation of hADSCs, and promoted the migration and invasion of hADSCs, as indicated by increased levels of MMP-2 and MMP-9. Furthermore, Netrin-1 overexpression increased the expression of vascular endothelial growth factor and placental growth factor in hADSCs. Our results highlighted the possibility that genetic modification of hADSCs by Netrin-1 overexpression might be beneficial for cell transplantation therapy against ischemic diseases.
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The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.
References
Ahmed RP, Haider KH, Shujia J, Afzal MR, Ashraf M (2010) Sonic Hedgehog gene delivery to the rodent heart promotes angiogenesis via iNOS/netrin-1/PKC pathway. PLoS One 5:e8576. https://doi.org/10.1371/journal.pone.0008576
Ballestín A, Casado JG, Abellán E, Vela FJ, Álvarez V, Usón A, Blázquez R, Sánchez-Margallo FM (2018) Adipose-derived stem cells ameliorate ischemia-reperfusion injury in a rat skin free flap model. J Reconstr Microsurg 34:601–609. https://doi.org/10.1055/s-0038-1648246
Biressi S, Filareto A, Rando TA (2020) Stem cell therapy for muscular dystrophies. J Clin Invest 130:5652–5664. https://doi.org/10.1172/JCI142031
Calabrese EJ (2021) Hormesis and adult adipose-derived stem cells. Pharmacol Res 172:105803. https://doi.org/10.1016/j.phrs.2021.105803
Charalambous F, Elia A, Georgiades P (2012) Decidual spiral artery remodeling during early post-implantation period in mice: investigation of associations with decidual uNK cells and invasive trophoblast. Biochem Biophys Res Commun 417:847–52. https://doi.org/10.1016/j.bbrc.2011.12.057
Colamarino SA, Tessier-Lavigne M (1995) The axonal chemoattractant netrin-1 is also a chemorepellent for trochlear motor axons. Cell 81:621–9. https://doi.org/10.1016/0092-8674(95)90083-7
Dakouane-Giudicelli M, Alfaidy N, de Mazancourt P (2014) Netrins and their roles in placental angiogenesis. Biomed Res Int 2014:901941. https://doi.org/10.1155/2014/901941
Dhanasekaran M, Indumathi S, Poojitha R, Kanmani A, Rajkumar JS, Sudarsanam D (2013) Plasticity and banking potential of cultured adipose tissue derived mesenchymal stem cells. Cell Tissue Bank 14:303–315. https://doi.org/10.1007/s10561-012-9311-7
Dun XP, Parkinson DB (2017) Role of Netrin-1 signaling in nerve regeneration. Int J Mol Sci 18:491. https://doi.org/10.3390/ijms18030491
Fisher SA, Doree C, Brunskill SJ, Mathur A, Martin-Rendon E (2013) Bone marrow stem cell treatment for ischemic heart disease in patients with no option of revascularization: a systematic review and meta-analysis. PLoS One 8:e64669. https://doi.org/10.1371/journal.pone.0064669
Frese L, Dijkman PE, Hoerstrup SP (2016) Adipose tissue-derived stem cells in regenerative medicine. Transfus Med Hemother 43:268–274. https://doi.org/10.1159/000448180
Han Y, Ren J, Bai Y, Pei X, Han Y (2019) Exosomes from hypoxia-treated human adipose-derived mesenchymal stem cells enhance angiogenesis through VEGF/VEGF-R. Int J Biochem Cell Biol 109:59–68. https://doi.org/10.1016/j.biocel.2019.01.017
Heidari-Moghadam A, Bayati V, Orazizadeh M, Rashno M (2020) Role of vascular endothelial growth factor and human umbilical vein endothelial cells in designing an in vitro vascular-muscle cellular model using adipose-derived stem cells. Cell J 22:19–28. https://doi.org/10.22074/CELLJ.2020.7034
Hou L, Kim JJ, Woo YJ, Huang NF (2016) Stem cell-based therapies to promote angiogenesis in ischemic cardiovascular disease. Am J Physiol Heart Circ Physiol 310:H455-65. https://doi.org/10.1152/ajpheart.00726.2015
Hu L, Pan J (2020) Adipose-derived stem cell therapy shows promising results for secondary lymphedema. World J Stem Cells 12:612–620. https://doi.org/10.4252/wjsc.v12.i7.612
Hwang OK, Noh YW, Hong JT, Lee JW (2020) Hypoxia pretreatment promotes chondrocyte differentiation of human adipose-derived stem cells via vascular endothelial growth factor. Tissue Eng Regen Med 17:335–350. https://doi.org/10.1007/s13770-020-00265-5
Jiao ZH, Liu XN, Ma YJ, Ge YS, Zhang QZ, Liu BY, Wang HB (2020a) Adipose-derived stem cells protect ischemia-reperfusion and partial hepatectomy by attenuating endoplasmic reticulum stress. Front Cell Dev Biol 8:177. https://doi.org/10.3389/fcell.2020.00177
Jiao X, Zhang D, Hong Q, Yan L, Han Q, Shao F, Cai G, Chen X, Zhu H (2020b) Netrin-1 works with UNC5B to regulate angiogenesis in diabetic kidney disease. Front Med 14:293–304. https://doi.org/10.1007/s11684-019-0715-7
Ke XJ, Liu CX, Wang Y, Ma JH, Mao XM, Li Q (2016) Netrin-1 promotes mesenchymal stem cell revascularization of limb ischaemia. Diab Vasc Dis Res 13:145–56. https://doi.org/10.1177/1479164115611594
Khazaei S, Keshavarz G, Bozorgi A, Nazari H, Khazaei M (2022) Adipose tissue-derived stem cells: a comparative review on isolation, culture, and differentiation methods. Cell Tissue Bank 23:1–16. https://doi.org/10.1007/s10561-021-09905-z
Koh YG, Kwon OR, Kim YS, Choi YJ, Tak DH (2016) Adipose-derived mesenchymal stem cells with microfracture versus microfracture alone: 2-Year follow-up of a prospective randomized trial. Arthroscopy 32:97–109. https://doi.org/10.1016/j.arthro.2015.09.010
Lee S, Chae DS, Song BW, Lim S, Kim SW, Kim IK, Hwang KC (2021) ADSC-Based cell therapies for musculoskeletal disorders: a review of recent clinical trials. Int J Mol Sci 22:10586. https://doi.org/10.3390/ijms221910586
Liu XX, Luo QQ, Zheng YF, Liu XP, Hu Y, Wang F, Zou L (2015) The role of Delta-like 4 ligand/Notch-ephrin-B2 cascade in the pathogenesis of preeclampsia by regulating functions of endothelial progenitor cell. Placenta 36:1002–10. https://doi.org/10.1016/j.placenta.2015.07.123
Llambi F, Causeret F, Bloch-Gallego E, Mehlen P (2001) Netrin-1 acts as a survival factor via its receptors UNC5H and DCC. EMBO J 20:2715–22. https://doi.org/10.1093/emboj/20.11.2715
Luo ML, Liu XP, Wang F, Liu XX, Liu WF, Wu D, Tao H, Wang RL, Zhao Y, Zhu JW, Zou L (2018) Conditioned medium from human umbilical vein endothelial cells promotes proliferation, migration, invasion and angiogenesis of adipose dirived stem cells. Curr Med Sci 38:124–130. https://doi.org/10.1007/s11596-018-1855-8
Mazo M, Hernandez S, Gavira JJ, Abizanda G, Araña M, López-Martínez T, Moreno C, Merino J, Martino-Rodríguez A, Uixeira A, de Jalón JAG, Pastrana J, Martínez-Caro D, Prósper F (2012) Treatment of reperfused ischemia with adipose-derived stem cells in a preclinical Swine model of myocardial infarction. Cell Transplant 21:2723–33. https://doi.org/10.3727/096368912X638847
Mori D, Miyagawa S, Yajima S, Saito S, Fukushima S, Ueno T, Toda K, Kawai K, Kurata H, Nishida H, Isohashi K, Hatazawa J, Sawa Y (2018) Cell spray transplantation of adipose-derived mesenchymal stem cell recovers ischemic cardiomyopathy in a porcine model. Transplantation 102:2012–2024. https://doi.org/10.1097/TP.0000000000002385
Müller P, Lemcke H, David R (2018) Stem cell therapy in heart diseases - cell types, mechanisms and improvement strategies. Cell Physiol Biochem 48:2607–2655. https://doi.org/10.1159/000492704
Park KW, Crouse D, Lee M, Karnik SK, Sorensen LK, Murphy KJ, Kuo CJ, Li DY (2004) The axonal attractant Netrin-1 is an angiogenic factor. Proc Natl Acad Sci USA 101:16210–5. https://doi.org/10.1073/pnas.0405984101
Pu LL (2012) Towards more rationalized approach to autologous fat grafting. J Plast Reconstr Aesthet Surg 65:413–9. https://doi.org/10.1016/j.bjps.2011.09.033
Sasagawa T, Shimizu T, Sekiya S, Yamato M, Okano T (2014) Comparison of angiogenic potential between prevascular and non-prevascular layered adipose-derived stem cell-sheets in early post-transplanted period. J Biomed Mater Res Part A 102:358–365. https://doi.org/10.1002/jbm.a.34707
Song L, Yang YJ, Dong QT, Qian HY, Gao RL, Qiao SB, Shen R, He ZX, Lu MJ, Zhao SH, Geng YJ, Gersh BJ (2013) Atorvastatin enhances efficacy of mesenchymal stem cells treatment for swine myocardial infarction via activation of nitric oxide synthase. PLoS One 8:e65702. https://doi.org/10.1371/journal.pone.0065702
Storck K, Ell J, Regn S, Rittler-Ungetüm B, Mayer H, Schantz T, Müller D, Buchberger M (2015) Optimization of in vitro cultivation strategies for human adipocyte derived stem cells. Adipocyte 4:181–187. https://doi.org/10.4161/21623945.2014.987580
Suresh V, West JL (2020) 3D culture facilitates VEGF-Stimulated endothelial differentiation of adipose-derived stem cells. Ann Biomed Eng 48:1034–1044. https://doi.org/10.1007/s10439-019-02297-y
Tucker HA, Bunnell BA (2011) Characterization of human adipose-derived stem cells using flow cytometry. Methods Mol Biol 702:121–31. https://doi.org/10.1007/978-1-61737-960-4_10
Volz AC, Huber B, Kluger PJ (2016) Adipose-derived stem cell differentiation as a basic tool for vascularized adipose tissue engineering. Differ Res Biol Div 92:52–64. https://doi.org/10.1016/j.diff.2016.02.003
Wilson BD, Ii M, Park KW, Suli A, Kock GH, Sorensen LK, Suh W, Larrieu-Lahargue F, Urness LD, Thomas KR, Chien CB, Losordo DW, Li DY (2006) Netrins promote developmental and therapeutic angiogenesis. Science 313:640–4. https://doi.org/10.1126/science.1124704
Wu HY, Peng Z, Xu Y, Sheng ZX, Liu YS, Liao YG, Wang Y, Wen Y, Yi JZ, Xie C, Chen XR, Hu JJ, Yan BQ, Wang HJ, Yao XD, Fu W, Ouyang HW (2022) Engineered adipose-derived stem cells with IGF-1-modified mRNA ameliorates osteoarthritis development. Stem Cell Res Ther 13:19. https://doi.org/10.1186/s13287-021-02695-x
Xie H, Zou L, Zhu J, Yang Y (2011) Effects of netrin-1 and netrin-1 knockdown on human umbilical vein endothelial cells and angiogenesis of rat placenta. Placenta 32:546–553. https://doi.org/10.1016/j.placenta.2011.04.003
Yang Y, Zou L, Wang Y, Xu KS, Zhang JX, Zhang JH (2007) Axon guidance cue Netrin-1 has dual function in angiogenesis. Cancer Biol Ther 6:743–8. https://doi.org/10.4161/cbt.6.5.3976
Yang S, Guo S, Tong S, Sun X (2019) Promoting osteogenic differentiation of human adipose-derived stem cells by altering the expression of exosomal miRNA. Stem Cells Int 2019:1351860. https://doi.org/10.1155/2019/1351860
Yang X, Sun H, Tang T, Zhang WC, Li Y (2021) Netrin-1 promotes retinoblastoma-associated angiogenesis. Ann Transl Med 9:1683. https://doi.org/10.21037/atm-21-5560
Yu H, Lu K, Zhu JY, Wang J (2017) Stem cell therapy for ischemic heart diseases. Br Med Bull 121:135–154. https://doi.org/10.1093/bmb/ldw059
Zhai Y, Wu W, Xi X, Yu R (2020) Adipose-derived stem cells promote proliferation and invasion in cervical cancer by targeting the HGF/c-MET pathway. Cancer Manag Res 12:11823–11832. https://doi.org/10.2147/CMAR.S277130
Zhang X, Qin JB, Wang X, Guo X, Liu JC, Wang XH, Wu XY, Lu XW, Li WM, Liu XB (2018) Netrin-1 improves adipose-derived stem cell proliferation, migration, and treatment effect in type 2 diabetic mice with sciatic denervation. Stem Cell Res Ther 9:285. https://doi.org/10.1186/s13287-018-1020-0
Zhang X, Jiang YH, Huang Q, Wu ZY, Pu HJ, Xu ZJ, Li B, Lu XW, Yang XR, Qin JB, Peng ZY (2021) Exosomes derived from adipose-derived stem cells overexpressing glyoxalase-1 protect endothelial cells and enhance angiogenesis in type 2 diabetic mice with limb ischemia. Stem Cell Res Ther 12:403. https://doi.org/10.1186/s13287-021-02475-7
Zhao Y, Gao J, Lu F (2013) Human adipose-derived stem cell adipogenesis induces paracrine regulation of the invasive ability of MCF-7 human breast cancer cells in vitro. Exp Ther Med 6:937–942. https://doi.org/10.3892/etm.2013.1237
Zhao Y, Zheng YF, Luo QQ, Yan T, Liu XX, Han L, Zou L (2014) Edaravone inhibits hypoxia-induced trophoblast-soluble Fms-like tyrosine kinase 1 expression: a possible therapeutic approach to preeclampsia. Placenta 35:476–82. https://doi.org/10.1016/j.placenta.2014.04.002
Zhu Y, Wu Y, Cheng J, Wang Q, Li Z, Wang Y, Wang D, Wang H, Zhang W, Ye J, Jiang H, Wang L (2018) Pharmacological activation of TAZ enhances osteogenic differentiation and bone formation of adipose-derived stem cells. Stem Cell Res Ther 9:53. https://doi.org/10.1186/s13287-018-0799-z
Acknowledgements
This project was supported by grants from the National Natural Science Foundation of China (Grant No. 81100428).
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National Natural Science Foundation of China, 81100428.
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M-LL and J-W Z: designed the study. M-LL: carried out most experiments and analysis of the data. J-WZ: carried out the molecular experiments. X-MG: wrote the article, gave useful suggestions and discussed the manuscript. The authors declare that there is no conflict of interest with any financial organization or corporation or individual that can inappropriately influence this work.
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Luo, Ml., Zhu, Jw. & Gao, Xm. Netrin-1 promotes the vasculogenic capacity of human adipose-derived stem cells. Cell Tissue Bank 24, 357–367 (2023). https://doi.org/10.1007/s10561-022-10038-0
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DOI: https://doi.org/10.1007/s10561-022-10038-0