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Rab27a Regulates Human Perivascular Adipose Progenitor Cell Differentiation

  • Joshua M. Boucher
  • Michael Robich
  • S. Spencer Scott
  • Xuehui Yang
  • Larisa Ryzhova
  • Jacqueline E. Turner
  • Ilka Pinz
  • Lucy Liaw
ORIGINAL ARTICLE

Abstract

Purpose

Perivascular adipose tissue (PVAT) surrounds blood vessels and regulates vascular tone through paracrine secretion of cytokines. During conditions promoting cardiometabolic dysfunction, such as obesity, cytokine secretion is altered towards a proinflammatory and proatherogenic profile. Despite the clinical implications for cardiovascular disease, studies addressing the biology of human PVAT remain limited. We are interested in characterizing the resident adipose progenitor cells (APCs) because of their potential role in PVAT expansion during obesity. We also focused on proteins regulating paracrine interactions, including the small GTPase Rab27a, which regulates protein trafficking and secretion.

Methods

PVAT from the ascending aorta was collected from patients with severe cardiovascular disease undergoing coronary artery bypass grafting (CABG). Freshly-isolated PVAT was digested and APC expanded in culture for characterizing progenitor markers, evaluating adipogenic potential and assessing the function(s) of Rab27a.

Results

Using flow cytometry, RT-PCR, and immunoblot, we characterized APC from human PVAT as negative for CD45 and CD31 and expressing CD73, CD105, and CD140A. These APCs differentiate into multilocular, UCP1-producing adipocytes in vitro. Rab27a was detected in interstitial cells of human PVAT in vivo and along F-actin tracks of PVAT-APC in vitro. Knockdown of Rab27a using siRNA in PVAT-APC prior to induction resulted in a marked reduction in lipid accumulation and reduced expression of adipogenic differentiation markers.

Conclusions

PVAT-APC from CABG donors express common adipocyte progenitor markers and differentiate into UCP1-containing adipocytes. Rab27a has an endogenous role in promoting the maturation of adipocytes from human PVAT-derived APC.

Keywords

Perivascular Adipose Cardiovascular Disease Rab27a Progenitor 

Notes

Funding

This research was supported by NIH grant R01HL141149 (L. Liaw, PI) and American Heart Association grant 17GRNT33670972 (L. Liaw, PI). JB was partially supported by a pilot project from NIH grant 5P30GM106391, which also supported the Progenitor Cell Analysis Core, which was used for flow cytometry (R. Friesel PI). This work was also supported by our Histopathology and Histomorphometry Core, which is supported by NIH grants P20GM121301 (L. Liaw, PI), P30GM106391 (R. Friesel, PI), and U54GM115516 (C. Rosen, PI).

Compliance with Ethical Standards

Conflicts of Interest

The authors declare that they have no conflict of interest.

Ethical Approval

This study utilized human tissue that was procured via our Maine Medical Center Biobank, which provides de-identified samples. This study was reviewed and deemed exempt by our Maine Medical Center Institutional Review Board. The BioBank protocols are in accordance with the ethical standards of our institution and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.

This article does not contain any studies with animals performed by any of the authors.

Informed consent

Informed consent was obtained from all individuals for de-identified use of their samples for research purposes.

References

  1. 1.
    Chang L, Villacorta L, Li R, Hamblin M, Xu W, Dou C, et al. Loss of perivascular adipose tissue on peroxisome proliferator-activated receptor-gamma deletion in smooth muscle cells impairs intravascular thermoregulation and enhances atherosclerosis. Circulation. 2012;126(9):1067–78.CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Greenstein AS, Khavandi K, Withers SB, Sonoyama K, Clancy O, Jeziorska M, et al. Local inflammation and hypoxia abolish the protective anticontractile properties of perivascular fat in obese patients. Circulation. 2009;119(12):1661–70.CrossRefPubMedGoogle Scholar
  3. 3.
    Mazurek T, Opolski G. Pericoronary adipose tissue: a novel therapeutic target in obesity-related coronary atherosclerosis. J Am Coll Nutr. 2015;34(3):244–54.CrossRefPubMedGoogle Scholar
  4. 4.
    Lu D, Wang W, Xia L, Xia P, Yan Y. Gene expression profiling reveals heterogeneity of perivascular adipose tissues surrounding coronary and internal thoracic arteries. Acta Biochim Biophys Sin. 2017;49(12):1075–82.CrossRefPubMedGoogle Scholar
  5. 5.
    Samano N, Geijer H, Liden M, Fremes S, Bodin L, Souza D. The no-touch saphenous vein for coronary artery bypass grafting maintains a patency, after 16 years, comparable to the left internal thoracic artery: a randomized trial. J Thorac Cardiovasc Surg. 2015;150(4):880–8.CrossRefPubMedGoogle Scholar
  6. 6.
    Pawliszak W, Kowalewski M, Raffa GM, Malvindi PG, Kowalkowska ME, Szwed KA, et al. Cerebrovascular events after no-touch off-pump coronary artery bypass grafting, conventional side-clamp off-pump coronary artery bypass, and proximal anastomotic devices: a meta-analysis. J Am Heart Assoc. 2016;5(2).Google Scholar
  7. 7.
    Lindroos B, Suuronen R, Miettinen S. The potential of adipose stem cells in regenerative medicine. Stem Cell Rev. 2011;7(2):269–91.CrossRefPubMedGoogle Scholar
  8. 8.
    Suzuki E, Fujita D, Takahashi M, Oba S, Nishimatsu H. Adipose tissue-derived stem cells as a therapeutic tool for cardiovascular disease. World J Cardiol. 2015;7(8):454–65.CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Traustadottir GA, Kosmina R, Sheikh SP, Jensen CH, Andersen DC. Preadipocytes proliferate and differentiate under the guidance of Delta-like 1 homolog (DLK1). Adipocyte. 2013;2(4):272–5.CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Cawthorn WP, Scheller EL, MacDougald OA. Adipose tissue stem cells meet preadipocyte commitment: going back to the future. J Lipid Res. 2012;53(2):227–46.CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Baer PC. Adipose-derived mesenchymal stromal/stem cells: an update on their phenotype in vivo and in vitro. World J Stem Cells. 2014;6(3):256–65.CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Hamid AA, Idrus RB, Saim AB, Sathappan S, Chua KH. Characterization of human adipose-derived stem cells and expression of chondrogenic genes during induction of cartilage differentiation. Clinics. 2012;67(2):99–106.CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Zimmerlin L, Donnenberg VS, Rubin JP, Donnenberg AD. Mesenchymal markers on human adipose stem/progenitor cells. Cytometry A. 2013;83(1):134–40.CrossRefPubMedGoogle Scholar
  14. 14.
    Lee YH, Petkova AP, Mottillo EP, Granneman JG. In vivo identification of bipotential adipocyte progenitors recruited by beta3-adrenoceptor activation and high-fat feeding. Cell Metab. 2012;15(4):480–91.CrossRefPubMedPubMedCentralGoogle Scholar
  15. 15.
    Beckenkamp LR, Souza LEB, Melo FUF, Thome CH, Magalhaes DAR, Palma PVB, et al. Comparative characterization of CD271(+) and CD271(−) subpopulations of CD34(+) human adipose-derived stromal cells. J Cell Biochem. 2018;119(5):3873–84.CrossRefPubMedGoogle Scholar
  16. 16.
    Rosen ED, MacDougald OA. Adipocyte differentiation from the inside out. Nat Rev Mol Cell Biol. 2006;7(12):885–96.CrossRefPubMedGoogle Scholar
  17. 17.
    Park KW, Halperin DS, Tontonoz P. Before they were fat: adipocyte progenitors. Cell Metab. 2008;8(6):454–7.CrossRefPubMedGoogle Scholar
  18. 18.
    Moon YS, Smas CM, Lee K, Villena JA, Kim KH, Yun EJ, et al. Mice lacking paternally expressed Pref-1/Dlk1 display growth retardation and accelerated adiposity. Mol Cell Biol. 2002;22(15):5585–92.CrossRefPubMedPubMedCentralGoogle Scholar
  19. 19.
    Maumus M, Sengenes C, Decaunes P, Zakaroff-Girard A, Bourlier V, Lafontan M, et al. Evidence of in situ proliferation of adult adipose tissue-derived progenitor cells: influence of fat mass microenvironment and growth. J Clin Endocrinol Metab. 2008;93(10):4098–106.CrossRefPubMedGoogle Scholar
  20. 20.
    Fukuda M. Rab27 effectors, pleiotropic regulators in secretory pathways. Traffic. 2013;14(9):949–63.CrossRefPubMedGoogle Scholar
  21. 21.
    Boucher JM, Clark RP, Chong DC, Citrin KM, Wylie LA, Bautch VL. Dynamic alterations in decoy VEGF receptor-1 stability regulate angiogenesis. Nat Commun. 2017;8:15699.CrossRefPubMedPubMedCentralGoogle Scholar
  22. 22.
    Kasai K, Ohara-Imaizumi M, Takahashi N, Mizutani S, Zhao S, Kikuta T, et al. Rab27a mediates the tight docking of insulin granules onto the plasma membrane during glucose stimulation. J Clin Invest. 2005;115(2):388–96.CrossRefPubMedPubMedCentralGoogle Scholar
  23. 23.
    Johnson JL, Brzezinska AA, Tolmachova T, Munafo DB, Ellis BA, Seabra MC, et al. Rab27a and Rab27b regulate neutrophil azurophilic granule exocytosis and NADPH oxidase activity by independent mechanisms. Traffic. 2010;11(4):533–47.CrossRefPubMedGoogle Scholar
  24. 24.
    Menasche G, Pastural E, Feldmann J, Certain S, Ersoy F, Dupuis S, et al. Mutations in RAB27A cause Griscelli syndrome associated with haemophagocytic syndrome. Nat Genet. 2000;25(2):173–6.CrossRefPubMedGoogle Scholar
  25. 25.
    Wilson SM, Yip R, Swing DA, O'Sullivan TN, Zhang Y, Novak EK, et al. A mutation in Rab27a causes the vesicle transport defects observed in ashen mice. Proc Natl Acad Sci U S A. 2000;97(14):7933–8.CrossRefPubMedPubMedCentralGoogle Scholar
  26. 26.
    Ostrowski M, Carmo NB, Krumeich S, Fanget I, Raposo G, Savina A, et al. Rab27a and Rab27b control different steps of the exosome secretion pathway. Nat Cell Biol. 2010;12(1):19–30. sup pp 11–13CrossRefPubMedGoogle Scholar
  27. 27.
    Shimada-Sugawara M, Sakai E, Okamoto K, Fukuda M, Izumi T, Yoshida N, et al. Rab27A regulates transport of cell surface receptors modulating multinucleation and lysosome-related organelles in osteoclasts. Sci Rep. 2015;5:9620.CrossRefPubMedPubMedCentralGoogle Scholar
  28. 28.
    Kapur SK, Dos-Anjos Vilaboa S, Llull R, Katz AJ. Adipose tissue and stem/progenitor cells: discovery and development. Clin Plast Surg. 2015;42(2):155–67.CrossRefPubMedGoogle Scholar
  29. 29.
    Gupta RK, Mepani RJ, Kleiner S, Lo JC, Khandekar MJ, Cohen P, et al. Zfp423 expression identifies committed preadipocytes and localizes to adipose endothelial and perivascular cells. Cell Metab. 2012;15(2):230–9.CrossRefPubMedPubMedCentralGoogle Scholar
  30. 30.
    Rim JS, Mynatt RL, Gawronska-Kozak B. Mesenchymal stem cells from the outer ear: a novel adult stem cell model system for the study of adipogenesis. FASEB J. 2005;19(9):1205–7.CrossRefPubMedGoogle Scholar
  31. 31.
    Kraus NA, Ehebauer F, Zapp B, Rudolphi B, Kraus BJ, Kraus D. Quantitative assessment of adipocyte differentiation in cell culture. Adipocyte. 2016;5(4):351–8.CrossRefPubMedPubMedCentralGoogle Scholar
  32. 32.
    Brown NK, Zhou Z, Zhang J, Zeng R, Wu J, Eitzman DT, et al. Perivascular adipose tissue in vascular function and disease: a review of current research and animal models. Aterioscler Thromb Vasc Biol. 2014;8:1621–30.CrossRefGoogle Scholar
  33. 33.
    Skinner JR, Harris LA, Shew TM, Abumrad NA, Wolins NE. Perilipin 1 moves between the fat droplet and the endoplasmic reticulum. Adipocyte. 2013;2(2):80–6.CrossRefPubMedPubMedCentralGoogle Scholar
  34. 34.
    Miao CY, Li ZY. The role of perivascular adipose tissue in vascular smooth muscle cell growth. Br J Pharmacol. 2012;165(3):643–58.CrossRefPubMedPubMedCentralGoogle Scholar
  35. 35.
    Akoumianakis I, Tarun A, Antoniades C. Perivascular adipose tissue as a regulator of vascular disease pathogenesis: identifying novel therapeutic targets. Br J Pharmacol. 2017;174(20):3411–24.CrossRefPubMedGoogle Scholar
  36. 36.
    Nosalski R, Guzik TJ. Perivascular adipose tissue inflammation in vascular disease. Br J Pharmacol. 2017;174(20):3496–513.CrossRefPubMedPubMedCentralGoogle Scholar
  37. 37.
    Fukuda M. Versatile role of Rab27 in membrane trafficking: focus on the Rab27 effector families. J Biochem. 2005;137(1):9–16.CrossRefPubMedGoogle Scholar
  38. 38.
    Tolmachova T, Anders R, Stinchcombe J, Bossi G, Griffiths GM, Huxley C, et al. A general role for Rab27a in secretory cells. Mol Biol Cell. 2004;15(1):332–44.CrossRefPubMedPubMedCentralGoogle Scholar
  39. 39.
    Feng F, Jiang Y, Lu H, Lu X, Wang S, Wang L, et al. Rab27A mediated by NF-kappaB promotes the stemness of colon cancer cells via up-regulation of cytokine secretion. Oncotarget. 2016;7(39):63342–51.CrossRefPubMedPubMedCentralGoogle Scholar
  40. 40.
    Hannah MJ, Hume AN, Arribas M, Williams R, Hewlett LJ, Seabra MC, et al. Weibel-Palade bodies recruit Rab27 by a content-driven, maturation-dependent mechanism that is independent of cell type. J Cell Sci. 2003;116(Pt 19):3939–48.CrossRefPubMedGoogle Scholar
  41. 41.
    Stinchcombe JC, Barral DC, Mules EH, Booth S, Hume AN, Machesky LM, et al. Rab27a is required for regulated secretion in cytotoxic T lymphocytes. J Cell Biol. 2001;152(4):825–34.CrossRefPubMedPubMedCentralGoogle Scholar
  42. 42.
    Desnos C, Schonn JS, Huet S, Tran VS, El-Amraoui A, Raposo G, et al. Rab27A and its effector MyRIP link secretory granules to F-actin and control their motion towards release sites. J Cell Biol. 2003;163(3):559–70.CrossRefPubMedPubMedCentralGoogle Scholar
  43. 43.
    Ohyama K, Matsumoto Y, Nishimiya K, Hao K, Tsuburaya R, Ota H, et al. Increased coronary perivascular adipose tissue volume in patients with vasospastic angina. Circ J. 2016;80(7):1653–6.CrossRefPubMedGoogle Scholar
  44. 44.
    Folkesson M, Vorkapic E, Gulbins E, Japtok L, Kleuser B, Welander M, et al. Inflammatory cells, ceramides, and expression of proteases in perivascular adipose tissue adjacent to human abdominal aortic aneurysms. J Vasc Surg. 2017;65(4):1171–1179. e1.CrossRefPubMedGoogle Scholar
  45. 45.
    Drosos I, Chalikias G, Pavlaki M, Kareli D, Epitropou G, Bougioukas G, et al. Differences between perivascular adipose tissue surrounding the heart and the internal mammary artery: possible role for the leptin-inflammation-fibrosis-hypoxia axis. Clin Res Cardiol. 2016;105(11):887–900.CrossRefPubMedGoogle Scholar
  46. 46.
    Xia N, Li H. The role of perivascular adipose tissue in obesity-induced vascular dysfunction. Br J Pharmacol. 2017;174(20):3425–42.CrossRefPubMedGoogle Scholar
  47. 47.
    Ozen G, Daci A, Norel X, Topal G. Human perivascular adipose tissue dysfunction as a cause of vascular disease: focus on vascular tone and wall remodeling. Eur J Pharmacol. 2015;766:16–24.CrossRefPubMedGoogle Scholar
  48. 48.
    Li Q, Qi LJ, Guo ZK, Li H, Zuo HB, Li NN. CD73+ adipose-derived mesenchymal stem cells possess higher potential to differentiate into cardiomyocytes in vitro. J Mol Histol. 2013;44(4):411–22.CrossRefPubMedGoogle Scholar
  49. 49.
    Rohban R, Pieber TR. Mesenchymal stem and progenitor cells in regeneration: tissue specificity and regenerative potential. Stem Cells Int. 2017;2017:5173732.CrossRefPubMedPubMedCentralGoogle Scholar
  50. 50.
    El-Badawy A, Amer M, Abdelbaset R, Sherif SN, Abo-Elela M, Ghallab YH, et al. Adipose stem cells display higher regenerative capacities and more adaptable electro-kinetic properties compared to bone marrow-derived mesenchymal stromal cells. Sci Rep. 2016;6:37801.CrossRefPubMedPubMedCentralGoogle Scholar
  51. 51.
    Feng F, Zhang J, Fan X, Yuan F, Jiang Y, Lv R, et al. Downregulation of Rab27A contributes to metformin-induced suppression of breast cancer stem cells. Oncol Lett. 2017;14(3):2947–53.CrossRefPubMedPubMedCentralGoogle Scholar
  52. 52.
    Sun Y, Bilan PJ, Liu Z, Klip A. Rab8A and Rab13 are activated by insulin and regulate GLUT4 translocation in muscle cells. Proc Natl Acad Sci U S A. 2010;107(46):19909–14.CrossRefPubMedPubMedCentralGoogle Scholar
  53. 53.
    Vazirani RP, Verma A, Sadacca LA, Buckman MS, Picatoste B, Beg M, et al. Disruption of adipose Rab10-dependent insulin signaling causes hepatic insulin resistance. Diabetes. 2016;65(6):1577–89.CrossRefPubMedPubMedCentralGoogle Scholar
  54. 54.
    Pulido MR, Diaz-Ruiz A, Jimenez-Gomez Y, Garcia-Navarro S, Gracia-Navarro F, Tinahones F, et al. Rab18 dynamics in adipocytes in relation to lipogenesis, lipolysis and obesity. PLoS One. 2011;6(7):e22931.CrossRefPubMedPubMedCentralGoogle Scholar
  55. 55.
    Kaddai V, Gonzalez T, Keslair F, Gremeaux T, Bonnafous S, Gugenheim J, et al. Rab4b is a small GTPase involved in the control of the glucose transporter GLUT4 localization in adipocyte. PLoS One. 2009;4(4):e5257.CrossRefPubMedPubMedCentralGoogle Scholar
  56. 56.
    Reed SE, Hodgson LR, Song S, May MT, Kelly EE, McCaffrey MW, et al. A role for Rab14 in the endocytic trafficking of GLUT4 in 3T3-L1 adipocytes. J Cell Sci. 2013;126(Pt 9):1931–41.CrossRefPubMedPubMedCentralGoogle Scholar
  57. 57.
    Chatterjee TK, Aronow BJ, Tong WS, Manka D, Tang Y, Bogdanov VY, et al. Human coronary artery perivascular adipocytes overexpress genes responsible for regulating vascular morphology, inflammation, and hemostasis. Physiol Genomics. 2013;45(16):697–709.CrossRefPubMedPubMedCentralGoogle Scholar

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© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Center for Molecular MedicineMaine Medical Center Research InstituteScarboroughUSA
  2. 2.Division of Thoracic and Cardiac SurgeryMaine Medical CenterPortlandUSA

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