Propranolol treatment of infantile hemangioma endothelial cells: A molecular analysis

Stiles,Jessica; Amaya,Clarissa; Pham,Robert; Rowntree,Rebecca K.; Lacaze,Mary; Mulne,Arlynn; Bischoff,Joyce; Kokta,Victor; Boucheron,Laura  E.; Mitchell,Dianne   C.; Bryan,Brad  A.

doi:10.3892/etm.2012.654

Propranolol treatment of infantile hemangioma endothelial cells: A molecular analysis

Authors:
- Jessica Stiles
- Clarissa Amaya
- Robert Pham
- Rebecca K. Rowntree
- Mary Lacaze
- Arlynn Mulne
- Joyce Bischoff
- Victor Kokta
- Laura E. Boucheron
- Dianne C. Mitchell
- Brad A. Bryan
View Affiliations

Affiliations: Department of Biomedical Sciences, Paul L. Foster School of Medicine, Texas Tech University Health Sciences Center, El Paso, TX, USA, Klipsch School of Electrical and Computer Engineering, New Mexico State University, Las Cruces, NM, USA , Department of Pediatrics, Paul L. Foster School of Medicine, Texas Tech University Health Sciences Center, El Paso, TX, USA, Vascular Biology Program and Department of Surgery, Children's Hospital Boston and Harvard Medical School, Boston, MA, USA, Department of Pathology, CHU Sainte-Justine, University of Montreal, Montreal, Quebec, Canada
Published online on: August 3, 2012 https://doi.org/10.3892/etm.2012.654
Pages: 594-604

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Infantile hemangiomas (IHs) are non-malignant, largely cutaneous vascular tumors affecting approximately 5-10% of children to varying degrees. During the first year of life, these tumors are strongly proliferative, reaching an average size ranging from 2 to 20 cm. These lesions subsequently stabilize, undergo a spontaneous slow involution and are fully regressed by 5 to 10 years of age. Systemic treatment of infants with the non-selective β-adrenergic receptor blocker, propranolol, has demonstrated remarkable efficacy in reducing the size and appearance of IHs. However, the mechanism by which this occurs is largely unknown. In this study, we sought to understand the molecular mechanisms underlying the effectiveness of β blocker treatment in IHs. Our data reveal that propranolol treatment of IH endothelial cells, as well as a panel of normal primary endothelial cells, blocks endothelial cell proliferation, migration, and formation of the actin cytoskeleton coincident with alterations in vascular endothelial growth factor receptor-2 (VEGFR-2), p38 and cofilin signaling. Moreover, propranolol induces major alterations in the protein levels of key cyclins and cyclin-dependent kinase inhibitors, and modulates global gene expression patterns with a particular affect on genes involved in lipid/sterol metabolism, cell cycle regulation, angiogenesis and ubiquitination. Interestingly, the effects of propranolol were endothelial cell-type independent, affecting the properties of IH endothelial cells at similar levels to that observed in neonatal dermal microvascular and coronary artery endothelial cells. This data suggests that while propranolol markedly inhibits hemangioma and normal endothelial cell function, its lack of endothelial cell specificity hints that the efficacy of this drug in the treatment of IHs may be more complex than simply blockage of endothelial function as previously believed.

View Figures

View References

1	Waner M, North PE, Scherer KA, Frieden IJ, Waner A and Mihm MC Jr: The nonrandom distribution of facial hemangiomas. Arch Dermatol. 139:869–875. 2003. View Article : Google Scholar : PubMed/NCBI
2	Léauté-Labrèze C, Dumas de la Roque E, Hubiche T, Boralevi F, Thambo JB and Taïeb A: Propranolol for severe hemangiomas of infancy. N Engl J Med. 358:2649–2651. 2008.PubMed/NCBI
3	D’Angelo G, Lee H and Weiner RI: cAMP-dependent protein kinase inhibits the mitogenic action of vascular endothelial growth factor and fibroblast growth factor in capillary endothelial cells by blocking Raf activation. J Cell Biochem. 67:353–366. 1997.
4	Sommers Smith SK and Smith DM: Beta blockade induces apoptosis in cultured capillary endothelial cells. In Vitro Cell Dev Biol Anim. 38:298–304. 2002.PubMed/NCBI
5	Lamy S, Lachambre MP, Lord-Dufour S and Béliveau R: Propranolol suppresses angiogenesis in vitro: inhibition of proliferation, migration, and differentiation of endothelial cells. Vascul Pharmacol. 53:200–208. 2010. View Article : Google Scholar : PubMed/NCBI
6	Spindler V and Waschke J: Beta-adrenergic stimulation contributes to maintenance of endothelial barrier functions under baseline conditions. Microcirculation. 18:118–127. 2011. View Article : Google Scholar
7	Makarski JS: Stimulation of cyclic AMP production by vasoactive agents in cultured bovine aortic and pulmonary artery endothelial cells. In Vitro. 17:450–458. 1981. View Article : Google Scholar : PubMed/NCBI
8	Welles SL, Shepro D and Hechtman HB: Vasoactive amines modulate actin cables (stress fibers) and surface area in cultured bovine endothelium. J Cell Physiol. 123:337–342. 1985. View Article : Google Scholar : PubMed/NCBI
9	Brehm BR, Bertsch D, von Fallois J and Wolf SC: Beta-blockers of the third generation inhibit endothelin-1 liberation, mRNA production and proliferation of human coronary smooth muscle and endothelial cells. J Cardiovasc Pharmacol. 36(Suppl 1): S401–S403. 2000. View Article : Google Scholar : PubMed/NCBI
10	Dai Y, Hou F, Buckmiller L, Fan CY, Saad A, Suen J and Richter GT: Decreased eNOS protein expression in involuting and propranolol-treated hemangiomas. Arch Otolaryngol Head Neck Surg. 138:177–182. 2012. View Article : Google Scholar : PubMed/NCBI
11	Chim H, Armijo BS, Miller E, Gliniak C, Serret MA and Gosain AK: Propranolol induces regression of hemangioma cells through HIF-1alpha-mediated inhibition of VEGF-A. Ann Surg. 256:146–156. 2012. View Article : Google Scholar : PubMed/NCBI
12	Boye E, Yu Y, Paranya G, Mulliken JB, Olsen BR and Bischoff J: Clonality and altered behavior of endothelial cells from hemangiomas. J Clin Invest. 107:745–752. 2001. View Article : Google Scholar : PubMed/NCBI
13	Lichtenstein N, Geiger B and Kam Z: Quantitative analysis of cytoskeletal organization by digital fluorescent microscopy. Cytometry A. 54:8–18. 2003. View Article : Google Scholar : PubMed/NCBI
14	Howell RE, Albelda SM, Daise ML and Levine EM: Characterization of beta-adrenergic receptors in cultured human and bovine endothelial cells. J Appl Physiol. 65:1251–1257. 1988.PubMed/NCBI
15	Jinnin M, Medici D, Park L, Limaye N, Liu Y, Boscolo E, Bischoff J, Vikkula M, Boye E and Olsen BR: Suppressed NFAT-dependent VEGFR1 expression and constitutive VEGFR2 signaling in infantile hemangioma. Nat Med. 14:1236–1246. 2008. View Article : Google Scholar : PubMed/NCBI
16	Souza BR, Santos JS and Costa AM: Blockade of beta1- and beta2-adrenoceptors delays wound contraction and re-epithelialization in rats. Clin Exp Pharmacol Physiol. 33:421–430. 2006. View Article : Google Scholar : PubMed/NCBI
17	Romana-Souza B, Santos JS and Monte-Alto-Costa A: beta-1 and beta-2, but not alpha-1 and alpha-2, adrenoceptor blockade delays rat cutaneous wound healing. Wound Repair Regen. 17:230–239. 2009. View Article : Google Scholar : PubMed/NCBI
18	Romana-Souza B, Porto LC and Monte-Alto-Costa A: Cutaneous wound healing of chronically stressed mice is improved through catecholamine blockade. Exp Dermatol. 19:821–829. 2010. View Article : Google Scholar : PubMed/NCBI
19	Romana-Souza B, Otranto M, Vieira AM, Filgueiras CC, Fierro IM and Monte-Alto-Costa A: Rotational stress-induced increase in epinephrine levels delays cutaneous wound healing in mice. Brain Behav Immun. 24:427–437. 2010. View Article : Google Scholar : PubMed/NCBI
20	Romana-Souza B, Nascimento AP and Monte-Alto-Costa A: Propranolol improves cutaneous wound healing in streptozotocin-induced diabetic rats. Eur J Pharmacol. 611:77–84. 2009. View Article : Google Scholar : PubMed/NCBI
21	Romana-Souza B, Nascimento AP and Monte-Alto-Costa A: Low-dose propranolol improves cutaneous wound healing of burn-injured rats. Plast Reconstr Surg. 122:1690–1699. 2008. View Article : Google Scholar : PubMed/NCBI
22	Mohammadi AA, Bakhshaeekia A, Alibeigi P, Hasheminasab MJ, Tolide-ei HR, Tavakkolian AR and Mohammadi MK: Efficacy of propranolol in wound healing for hospitalized burn patients. J Burn Care Res. 30:1013–1017. 2009.PubMed/NCBI
23	Musumeci M, Maccari S, Sestili P, Signore M, Molinari P, Ambrosio C, Stati T, Colledge WH, Grace AA, Catalano L and Marano G: Propranolol enhances cell cycle-related gene expression in pressure overloaded hearts. Br J Pharmacol. 164:1917–1928. 2011. View Article : Google Scholar : PubMed/NCBI
24	Erraji-Benchekroun L, Couton D, Postic C, Borde I, Gaston J, Guillet JG and André C: Overexpression of beta2-adrenergic receptors in mouse liver alters the expression of gluconeogenic and glycolytic enzymes. Am J Physiol Endocrinol Metab. 288:E715–E722. 2005. View Article : Google Scholar : PubMed/NCBI
25	Patrizio M, Musumeci M, Stati T, Fecchi K, Mattei E, Catalano L and Marano G: Propranolol promotes Egr1 gene expression in cardiomyocytes via beta-adrenoceptors. Eur J Pharmacol. 587:85–89. 2008. View Article : Google Scholar : PubMed/NCBI
26	Lashbrook BL and Steinle JJ: Beta-adrenergic receptor regulation of pigment epithelial-derived factor expression in rat retina. Auton Neurosci. 121:33–39. 2005. View Article : Google Scholar : PubMed/NCBI
27	Haggstrom AN, Beaumont JL, Lai JS, Adams DM, Drolet BA, Frieden IJ, Garzon MC, Holland KE, Horii KA, Lucky AW, et al: Measuring the severity of infantile hemangiomas: instrument development and reliability. Arch Dermatol. 148:197–202. 2012. View Article : Google Scholar : PubMed/NCBI
28	Boye E and Olsen BR: Signaling mechanisms in infantile hemangioma. Curr Opin Hematol. 16:202–208. 2009. View Article : Google Scholar : PubMed/NCBI
29	Grimmer JF, Williams MS, Pimentel R, Mineau G, Wood GM, Bayrak-Toydemir P and Stevenson DA: Familial clustering of hemangiomas. Arch Otolaryngol Head Neck Surg. 137:757–760. 2011. View Article : Google Scholar : PubMed/NCBI
30	Khan ZA, Boscolo E, Picard A, Psutka S, Melero-Martin JM, Bartch TC, Mulliken JB and Bischoff J: Multipotential stem cells recapitulate human infantile hemangioma in immunodeficient mice. J Clin Invest. 118:2592–2599. 2008.PubMed/NCBI
31	Calicchio ML, Collins T and Kozakewich HP: Identification of signaling systems in proliferating and involuting phase infantile hemangiomas by genome-wide transcriptional profiling. Am J Pathol. 174:1638–1649. 2009. View Article : Google Scholar : PubMed/NCBI
32	Iaccarino G, Ciccarelli M, Sorriento D, Galasso G, Campanile A, Santulli G, Cipolletta E, Cerullo V, Cimini V, Altobelli GG, et al: Ischemic neoangiogenesis enhanced by beta2-adrenergic receptor overexpression: a novel role for the endothelial adrenergic system. Circ Res. 97:1182–1189. 2005. View Article : Google Scholar
33	Storch CH and Hoeger PH: Propranolol for infantile haemangiomas: insights into the molecular mechanisms of action. Br J Dermatol. 163:269–274. 2010. View Article : Google Scholar : PubMed/NCBI
34	Fredriksson JM, Lindquist JM, Bronnikov GE and Nedergaard J: Norepinephrine induces vascular endothelial growth factor gene expression in brown adipocytes through a beta-adrenoreceptor/cAMP/protein kinase A pathway involving Src but independently of Erk1/2. J Biol Chem. 275:13802–13811. 2000. View Article : Google Scholar
35	Taylor MR: Pharmacogenetics of the human beta-adrenergic receptors. Pharmacogenomics J. 7:29–37. 2007. View Article : Google Scholar : PubMed/NCBI
36	Rohrer DK, Chruscinski A, Schauble EH, Bernstein D and Kobilka BK: Cardiovascular and metabolic alterations in mice lacking both beta1- and beta2-adrenergic receptors. J Biol Chem. 274:16701–16708. 1999. View Article : Google Scholar : PubMed/NCBI
37	Krief S, Lönnqvist F, Raimbault S, Baude B, Van Spronsen A, Arner P, Strosberg AD, Ricquier D and Emorine LJ: Tissue distribution of beta 3-adrenergic receptor mRNA in man. J Clin Invest. 91:344–349. 1993. View Article : Google Scholar : PubMed/NCBI
38	Yuan SM, Jiang HQ, Ouyang TX and Xing X: The distribution and evolution of pericytes in infantile hemangioma. Zhonghua Zheng Xing Wai Ke Za Zhi. 23:322–324. 2007.(In Chinese).
39	Ferrari-Dileo G, Davis EB and Anderson DR: Effects of cholinergic and adrenergic agonists on adenylate cyclase activity of retinal microvascular pericytes in culture. Invest Ophthalmol Vis Sci. 33:42–47. 1992.
40	Siegfried EC, Keenan WJ and Al-Jureidini S: More on propranolol for hemangiomas of infancy. N Engl J Med. 359:2846author reply 2846–2847, 2008.
41	Thaker PH, Han LY, Kamat AA, Arevalo JM, Takahashi R, Lu C, Jennings NB, Armaiz-Pena G, Bankson JA, Ravoori M, et al: Chronic stress promotes tumor growth and angiogenesis in a mouse model of ovarian carcinoma. Nat Med. 12:939–944. 2006. View Article : Google Scholar : PubMed/NCBI
42	Yang H, Xu Z, Iuvone PM and Grossniklaus HE: Angiostatin decreases cell migration and vascular endothelium growth factor (VEGF) to pigment epithelium derived factor (PEDF) RNA ratio in vitro and in a murine ocular melanoma model. Mol Vis. 12:511–517. 2006.
43	Schieven GL: The biology of p38 kinase: a central role in inflammation. Curr Top Med Chem. 5:921–928. 2005. View Article : Google Scholar : PubMed/NCBI
44	Bamburg JR and Wiggan OP: ADF/cofilin and actin dynamics in disease. Trends Cell Biol. 12:598–605. 2002. View Article : Google Scholar : PubMed/NCBI
45	Arber S, Barbayannis FA, Hanser H, Schneider C, Stanyon CA, Bernard O and Caroni P: Regulation of actin dynamics through phosphorylation of cofilin by LIM-kinase. Nature. 393:805–809. 1998. View Article : Google Scholar : PubMed/NCBI
46	Street CA and Bryan BA: Rho kinase proteins – pleiotropic modulators of cell survival and apoptosis. Anticancer Res. 31:3645–3657. 2011.
47	De Giorgi V, Gandini S, Grazzini M, Benemei S, Marchionni N and Geppetti P: Beta-blockers: a new and emerging treatment for melanoma. Recenti Prog Med. 103:11–16. 2012.(In Italian).
48	Lin X, Luo K, Lv Z and Huang J: Beta-adrenoceptor action on pancreatic cancer cell proliferation and tumor growth in mice. Hepatogastroenterology. 59:584–588. 2012.PubMed/NCBI
49	Jansen L, Below J, Chang-Claude J, Brenner H and Hoffmeister M: Beta blocker use and colorectal cancer risk: Population-based case-control study. Cancer. May 14–2012.(Epub ahead of print).
50	Pérez Piñero C, Bruzzone A, Sarappa MG, Castillo LF and Lüthy IA: Involvement of alpha2- and beta2-adrenoceptors on breast cancer cell proliferation and tumour growth regulation. Br J Pharmacol. 166:721–736. 2012.PubMed/NCBI
51	Chambers CB, Katowitz WR, Katowitz JA and Binenbaum G: A controlled study of topical 0.25% timolol maleate gel for the treatment of cutaneous infantile capillary hemangiomas. Ophthal Plast Reconstr Surg. 28:103–106. 2012.