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Cytoskeletal Priming of Mesenchymal Stem Cells to a Medicinal Phenotype

Regenerative Engineering and Translational Medicine volume 3, pages 5–14 (2017)Cite this article

Abstract

Mesenchymal stem cell (MSC) therapy is a promising approach for the treatment of cardiovascular disease, demonstrating pronounced trophic, immunomodulatory, and pro-angiogenic activity. However, clinical efficacy has suffered from broad variability, presumably due to cell death upon implantation, and the heterogeneous population of autologous cells. Micropatterning single cells in the same geometry can normalize the phenotype in a population, and variations in subcellular curvature will guide focal adhesion, cytoskeletal organization, and the regulation of distinct epigenetic marks to orchestrate a medicinal secretome. Within 2 days, activated cells show elevated expression of pericyte markers and will recapitulate functional pericyte activity through enhanced association with endothelial cell tubules in co-culture. MSCs are believed to undergo a temporary switch in vivo to an activated state in response to injury; thus, we propose engineering actomyosin contractility after isolation can similarly activate MSCs, which may serve as a general approach to prime a medicinal phenotype for cell-based therapies.

Lay Summary

Patient-derived mesenchymal stem cells will secrete molecules that promote new vasculature and have demonstrated clinical efficacy as a therapy for treating myocardial infarction. While autologous cell implantation is promising, there is considerable variability in current treatments and controversy over the underlying mechanisms. In this work, we reveal that careful control of actomyosin contractility in mesenchymal stem cells can prime a pericyte state with a medicinal secretome that shows sustained pro-angiogenic activity in vitro and in vivo. Activation of mesenchymal stem cells to a medicinal phenotype may increase the efficacy and reproducibility of cell-based therapies for cardiovascular disease

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References

  1. Mozaffarian D, Benjamin EJ, Go AS, Arnett DK, Blaha MJ, Cushman M, et al. Heart disease and stroke statistics—2015 update: a report from the American Heart Association. Circulation. 2015;131;e29–322.

  2. Deveza L, Choi J, Yang F. Therapeutic angiogenesis for treating cardiovascular diseases. Theranostics. 2012;2:801–14.

    Article  Google Scholar 

  3. Zachary I, Morgan RD. Therapeutic angiogenesis for cardiovascular disease: biological context, challenges, prospects. Heart. 2011;97:181–9.

    Article  Google Scholar 

  4. Carmeliet P, Jain RK. Molecular mechanisms and clinical applications of angiogenesis. Nature. 2011;473:298–307.

    Article  Google Scholar 

  5. Epstein SE, Kornowski R, Fuchs S, Dvorak HF. Angiogenesis therapy: amidst the hype, the neglected potential for serious side effects. Circulation. 2001;104:115–9.

    Article  Google Scholar 

  6. Beohar N, Rapp J, Pandya S, Losordo DW. Rebuilding the damaged heart: the potential of cytokines and growth factors in the treatment of ischemic heart disease. J Am Coll Cardiol. 2010;56:1287–97.

    Article  Google Scholar 

  7. Ranganath SH, Levy O, Inamdar MS, Karp JM. Harnessing the mesenchymal stem cell secretome for the treatment of cardiovascular disease. Cell Stem Cell. 2012;10:244–58.

    Article  Google Scholar 

  8. Pittenger MF, Mackay AM, Beck SC, Jaiswal RK, Douglas R, Mosca JD, et al. Multilineage potential of adult human mesenchymal stem cells. Science. 1999;284:143–7.

    Article  Google Scholar 

  9. Wingate K, Bonani W, Tan Y, Bryant SJ, Tan W. Compressive elasticity of three-dimensional nanofiber matrix directs mesenchymal stem cell differentiation to vascular cells with endothelial or smooth muscle cell markers. Acta Biomater. 2012;8:1440–9.

    Article  Google Scholar 

  10. Pijnappels DA, Schalij MJ, Ramkisoensing AA, Van Tuyn J, De Vries AA, Van Der Laarse A, et al. Forced alignment of mesenchymal stem cells undergoing cardiomyogenic differentiation affects functional integration with cardiomyocyte cultures. Circ Res. 2008;103:167–76.

    Article  Google Scholar 

  11. Leiker M. Assessment of a nuclear affinity labeling method for tracking implanted mesenchymal stem cells. Cell Transplant. 2008;17:911–22.

    Article  Google Scholar 

  12. Ankrum J, Karp JM. Mesenchymal stem cell therapy: two steps forward, one step back. Trends Mol Med. 2010;16:203–9.

    Article  Google Scholar 

  13. Crisan M, Yap S, Casteilla L, Chen C-W, Corselli M, Park TS, et al. A perivascular origin for mesenchymal stem cells in multiple human organs. Cell Stem Cell. 2008;3:301–13.

    Article  Google Scholar 

  14. Traktuev DO, Merfeld-Clauss S, Li J, Kolonin M, Arap W, Pasqualini R, et al. A population of multipotent CD34-positive adipose stromal cells share pericyte and mesenchymal surface markers, reside in a periendothelial location, and stabilize endothelial networks. Circ Res. 2008;102:77–85.

    Article  Google Scholar 

  15. Caplan AI. All MSCs are pericytes? Cell Stem Cell. 2008;3:229–30.

    Article  Google Scholar 

  16. Mills SJ, Cowin AJ, Kaur P. Pericytes, mesenchymal stem cells and the wound healing process. Cells. 2013;2:621–34.

    Article  Google Scholar 

  17. da Silva ML, Caplan AI, Nardi NB. In search of the in vivo identity of mesenchymal stem cells. Stem Cells. 2008;26:2287–99.

    Article  Google Scholar 

  18. Koike N, Fukumura D, Gralla O, Au P, Schechner JS, Jain RK. Tissue engineering: creation of long-lasting blood vessels. Nature. 2004;428:138–9.

    Article  Google Scholar 

  19. Engler AJ, Sen S, Sweeney HL, Discher DE. Matrix elasticity directs stem cell lineage specification. Cell. 2006;126:677–89.

    Article  Google Scholar 

  20. Lee J, Abdeen AA, Zhang D, Kilian KA. Directing stem cell fate on hydrogel substrates by controlling cell geometry, matrix mechanics and adhesion ligand composition. Biomaterials. 2013;34:8140–8.

    Article  Google Scholar 

  21. Li Y, Kilian KA. Bridging the gap: from 2D cell culture to 3D microengineered extracellular matrices. Adv Healthc Mater. 2015;4:2780–96.

    Article  Google Scholar 

  22. Kilian KA, Bugarija B, Lahn BT, Mrksich M. Geometric cues for directing the differentiation of mesenchymal stem cells. Proc Natl Acad Sci. 2010;107:4872–7.

    Article  Google Scholar 

  23. Zhang D, Kilian KA. The effect of mesenchymal stem cell shape on the maintenance of multipotency. Biomaterials. 2013;34:3962–9.

    Article  Google Scholar 

  24. Abdeen AA, Weiss JB, Lee J, Kilian KA. Matrix composition and mechanics direct proangiogenic signaling from mesenchymal stem cells. Tissue Eng Part A. 2014;20:2737–45.

    Article  Google Scholar 

  25. Krock BL, Skuli N, Simon MC. Hypoxia-induced angiogenesis: good and evil. Genes Cancer. 2011;2:1117–33.

    Article  Google Scholar 

  26. Johnson KE, Wilgus TA. Vascular endothelial growth factor and angiogenesis in the regulation of cutaneous wound repair. Adv. wound care. 2014;3:647–61.

    Article  Google Scholar 

  27. Mammoto A, Connor KM, Mammoto T, Yung CW, Huh D, Aderman CM, et al. A mechanosensitive transcriptional mechanism that controls angiogenesis. Nature. 2009;457:1103–8.

    Article  Google Scholar 

  28. Chen W, Frangogiannis NG. Fibroblasts in post-infarction inflammation and cardiac repair. Biochim Biophys Acta - Mol Cell Res. 1833;2013:945–53.

    Google Scholar 

  29. Darby I, Skalli O, Gabbiani G. α-smooth muscle actin is transiently expressed by myofibroblasts during experimental wound healing. Lab Investig. 63:21–9.

  30. Kutcher ME, Kolyada AY, Surks HK, Herman IM. Pericyte Rho GTPase mediates both pericyte contractile phenotype and capillary endothelial growth state. Am J Pathol American Society for Investigative Pathology. 2007;171:693–701.

    Google Scholar 

  31. Durham JT, Surks HK, Dulmovits BM, Herman IM. Pericyte contractility controls endothelial cell cycle progression and sprouting: insights into angiogenic switch mechanics. AJP Cell Physiol. 2014;307:C878–92.

    Article  Google Scholar 

  32. Théry M. Micropatterning as a tool to decipher cell morphogenesis and functions. J Cell Sci. 2010;123:4201–13.

    Article  Google Scholar 

  33. Lee J, Abdeen AA, Tang X, Saif TA, Kilian KA. Geometric guidance of integrin mediated traction stress during stem cell differentiation. Biomaterials. 2015;69:174–83.

    Article  Google Scholar 

  34. Kelly-goss MR, Sweat RS, Stapor PC, Peirce SM, Murfee WL. Targeting pericytes for angiogenic therapies. Microcirculation. 2014;21:345–57.

    Article  Google Scholar 

  35. Mitchell TS, Bradley J, Robinson GS, Shima DT, Ng YS. RGS5 expression is a quantitative measure of pericyte coverage of blood vessels. Angiogenesis. 2008;11:141–51.

    Article  Google Scholar 

  36. Berger M, Bergers G, Arnold B, Hammerling GJ, Ganss R. Regulator of G-protein signaling-5 induction in pericytes coincides with active vessel remodeling during neovascularization. Blood. 2005;105:1094–101.

    Article  Google Scholar 

  37. Blocki A, Wang Y, Koch M, Peh P, Beyer S, Law P, et al. Not all MSCs can act as pericytes: functional in vitro assays to distinguish pericytes from other mesenchymal stem cells in angiogenesis. Stem Cells Dev. 2013;22:2347–55.

    Article  Google Scholar 

  38. Shi YU, Li H, Zhang X, Fu Y, Huang YAN, Po P, et al. Continuous cyclic mechanical tension inhibited Runx2 expression in mesenchymal stem cells through RhoA-ERK1/2 pathway. J Cell Physiol. 2011;226:2159–69.

    Article  Google Scholar 

  39. Ponce ML, Kleinmann HK. The chick chorioallantoic membrane as an in vivo angiogenesis model. Curr. Protoc. Cell Biol. 2003;Chapter 19:Unit 19.5.

  40. Wang N, Tytell JD, Ingber DE. Mechanotransduction at a distance: mechanically coupling the extracellular matrix with the nucleus. Nat Rev Mol Cell Biol. 2009;10:75–82.

    Article  Google Scholar 

  41. Versaevel M, Grevesse T, Gabriele S. Spatial coordination between cell and nuclear shape within micropatterned endothelial cells. Nat Commun. 2012;3:671.

    Article  Google Scholar 

  42. Jain N, Iyer KV, Kumar A, Shivashankar GV. Cell geometric constraints induce modular gene-expression patterns via redistribution of HDAC3 regulated by actomyosin contractility. Proc Natl Acad Sci. 2013;110:3–8.

    Article  Google Scholar 

  43. Le Beyec J, Xu R, Lee S-Y, Nelson CM, Rizki A, Alcaraz J, et al. Cell shape regulates global histone acetylation in human mammary epithelial cells. Exp Cell Res. 2007;313:3066–75.

    Article  Google Scholar 

  44. Struhl K. Histone acetylation and transcriptional regulatory mechanisms. Genes Dev. 1998;12:599–606.

    Article  Google Scholar 

  45. Peters AHFM, O’Carroll D, Scherthan H, Mechtler K, Sauer S, Schöfer C, et al. Loss of the Suv39h histone methyltransferases impairs mammalian heterochromatin and genome stability. Cell. 2001;107:323–37.

    Article  Google Scholar 

  46. Karmodiya K, Krebs AR, Oulad-Abdelghani M, Kimura H, Tora L. H3K9 and H3K14 acetylation co-occur at many gene regulatory elements, while H3K14ac marks a subset of inactive inducible promoters in mouse embryonic stem cells. BMC Genomics. 2012;13:424.

    Article  Google Scholar 

  47. Rao B, Shibata Y, Strahl BD, Lieb JD. Dimethylation of histone H3 at lysine 36 demarcates regulatory and nonregulatory chromatin genome-wide. Mol Cell Biol. 2005;25:9447–59.

    Article  Google Scholar 

  48. Krejcí J, Uhlírová R, Galiová G, Kozubek S, Smigová J, Bártová E. Genome-wide reduction in H3K9 acetylation during human embryonic stem cell differentiation. J Cell Physiol. 2009;219:677–87.

    Article  Google Scholar 

  49. Seib FP, Prewitz M, Werner C, Bornhäuser M. Matrix elasticity regulates the secretory profile of human bone marrow-derived multipotent mesenchymal stromal cells (MSCs). Biochem Biophys Res Commun. 2009;389:663–7.

    Article  Google Scholar 

  50. Rustad KC, Gurtner GC. Mesenchymal stem cells home to sites of injury and inflammation. Adv Wound Care. 2012;1:147–52.

    Article  Google Scholar 

  51. Murphy MB, Moncivais K, Caplan AI. Mesenchymal stem cells: environmentally responsive therapeutics for regenerative medicine. Exp Mol Med. 2013;45:e54.

    Article  Google Scholar 

  52. Caplan AI, Correa D. The MSC: an injury drugstore. Cell Stem Cell. 2011;9:11–5.

    Article  Google Scholar 

  53. Stratman AN, Malotte KM, Mahan RD, Davis MJ, Davis GE. Pericyte recruitment during vasculogenic tube assembly stimulates endothelial basement membrane matrix formation. Blood. 2009;114:5091–101.

    Article  Google Scholar 

  54. Díaz-Flores L, Gutiérrez R, Madrid JF, Varela H, Valladares F, Acosta E, et al. Pericytes. Morphofunction, interactions and pathology in a quiescent and activated mesenchymal cell niche. Histol Histopathol. 2009;24:909–69.

    Google Scholar 

  55. Merino-gonzález C, Zuñiga FA, Escudero C, Ormazabal V, Reyes C, Nova-lamperti E, et al. Mesenchymal stem cell-derived extracellular vesicles promote angiogenesis: potential clinical application. Front Physiol. 2016;7:1–9.

    Article  Google Scholar 

  56. DePina AS, Langford GM. Vesicle transport: the role of actin filaments and myosin. Microsc Res Tech. 1999;47(2):93–106.

    Article  Google Scholar 

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Acknowledgements

This work was supported by the National Heart Lung and Blood Institute of the National Institutes of Health, grant number HL121757. The authors declare no competing financial interests. We would like to thank the Beckman Institute and Institute of Genomic Biology Imaging facilities.

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Authors and Affiliations

  1. Department of Materials Science and Engineering, University of Illinois at Urbana-Champaign, Urbana, IL, 61801, USA

    Amr A. Abdeen, Junmin Lee & Kristopher A. Kilian

  2. Department of Bioengineering, University of Illinois at Urbana-Champaign, Urbana, IL, 61801, USA

    Yanfen Li & Kristopher A. Kilian

Authors
  1. Amr A. Abdeen
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  2. Junmin Lee
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  4. Kristopher A. Kilian
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Correspondence to Kristopher A. Kilian.

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A.A.A. and J.L. contributed equally to this work.

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Abdeen, A.A., Lee, J., Li, Y. et al. Cytoskeletal Priming of Mesenchymal Stem Cells to a Medicinal Phenotype. Regen. Eng. Transl. Med. 3, 5–14 (2017). https://doi.org/10.1007/s40883-016-0021-8

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  • DOI: https://doi.org/10.1007/s40883-016-0021-8

Keywords

  • Mesenchymal stem cells
  • Micropatterning
  • Actomysin conctractility
  • Pericyte
  • Angiogenesis