Heparanase modulates Shh and Wnt3a signaling in human medulloblastoma cells

Ridgway,Lon  D.; Wetzel,Michael  D.; Marchetti,Dario

doi:10.3892/etm.2010.189

Heparanase modulates Shh and Wnt3a signaling in human medulloblastoma cells

Authors:
- Lon D. Ridgway
- Michael D. Wetzel
- Dario Marchetti
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Affiliations: Department of Pathology and Immunology, Baylor College of Medicine, Houston, TX 77030, USA, Department of Pathology and Immunology, Baylor College of Medicine, BCM - Taub Bldg., Suite T240A, Mail stop 315, One Baylor Plaza, Houston, TX 77030, USA
Published online on: December 28, 2010 https://doi.org/10.3892/etm.2010.189
Pages: 229-237

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Abstract

The pathogenesis of medulloblastoma (MB), the most common and aggressive brain tumor in children, is poorly understood. MB tumors respond to factors secreted by cerebellar Purkinje neurons, such as Sonic hedgehog (Shh) and Wingless-type MMTV integration site family member 3a (Wnt3a). Understanding the modulation of Shh/Wnt signaling is critical for the development of new MB treatments. Shh and Wnt3a induce MB cell proliferation and bind heparan sulfate glycosaminoglycan chains (HS-GAGs). HS-GAGs are components of syndecans, cell surface HS proteoglycans (HSPGs) which act as co-receptors for extracellular matrix-based ligands, and are targets of heparanase (HPSE). We hypothesized that extracellular HPSE activity modulates MB intracellular signaling of Shh/Wnt3a, involving syndecan-1 and -4 carboxy terminal-associated proteins and downstream targets. We compared the regulation of Shh/Wnt3a signaling subsequent to treatment with exogenous human active HPSE in MB lines possessing increased invasive abilities. We identified guanine nucleotide exchange factor-H1 (GEF-H1), a small GTPase guanine nucleotide exchange factor, as a new component of a syndecan signaling complex. Secondly, we demonstrated that HPSE modulated Shh/Wnt3-dependent expression and the intracellular distribution of GEF-H1, β-catenin and N-Myc. Thirdly, HPSE modulated Shh/Wnt3a-dependent gene expression of heparan sulfate proteoglycan (HSPG) and Gli transcription factors. Fourthly, pre-treatment with HPSE, alone or prior to Shh/Wnt3a exposure, altered small GTPase (Rac1/RhoA) activities differentially and promoted RhoA activation. Finally, the differential regulation of Rac1/RhoA activities by HPSE affected MB cell proliferation and invasion. Our results indicate that the HPSE/HSPG axis is implicated in critical MB cell signaling pathways with potential relevance for MB treatment.

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1.	Guessous F, Li Y and Abounader R: Signaling pathways in medulloblastoma. J Cell Physiol. 217:577–583. 2008. View Article : Google Scholar
2.	Flora A, Klisch TJ, Schuster G and Zoghbi HY: Deletion of Atoh1 disrupts Sonic Hedgehog signaling in the developing cerebellum and prevents medulloblastoma. Science. 326:1424–1427. 2009. View Article : Google Scholar : PubMed/NCBI
3.	Ribi K, Relly C, Landolt MA, Alber FD, Boltshauser E and Grotzer MA: Outcome of medulloblastoma in children: long-term complications and quality of life. Neuropediatrics. 36:357–365. 2005. View Article : Google Scholar : PubMed/NCBI
4.	Marchetti D, Mrak RE, Paulsen DD and Sinnappah-Kang ND: Neurotrophin receptors and heparanase: a functional axis in human medulloblastoma invasion. J Exp Clin Cancer Res. 26:5–23. 2007.PubMed/NCBI
5.	Gottardo NG and Gajjar A: Current therapy for medulloblastoma. Curr Treat Options Neurol. 8:319–334. 2006. View Article : Google Scholar
6.	Yauch RL, Dijkgraaf GJ, Alicke B, et al: Smoothened mutation confers resistance to a hedgehog pathway inhibitor in medulloblastoma. Science. 326:572–574. 2009. View Article : Google Scholar : PubMed/NCBI
7.	Vaillant C and Monard D: SHH pathway and cerebellar development. Cerebellum. 8:291–301. 2009. View Article : Google Scholar : PubMed/NCBI
8.	Cool SM and Nurcombe V: Heparan sulfate regulation of progenitor cell fate. J Cell Biochem. 99:1040–1051. 2006. View Article : Google Scholar : PubMed/NCBI
9.	Rubin JB, Choi Y and Segal RA: Cerebellar proteoglycans regulate sonic hedgehog responses during development. Development. 129:2223–2232. 2002.PubMed/NCBI
10.	Mythreye K and Blobe GC: Proteoglycan signaling co-receptors: roles in cell adhesion, migration and invasion. Cell Signal. 21:1548–1558. 2009. View Article : Google Scholar : PubMed/NCBI
11.	O'Connell MP, Fiori JL, Kershner EK, et al: Heparan sulfate proteoglycan modulation of Wnt5A signal transduction in meta-static melanoma cells. J Biol Chem. 284:28704–28712. 2009.PubMed/NCBI
12.	Iozzo RV: Heparan sulfate proteoglycans: intricate molecules with intriguing functions. J Clin Invest. 108:165–167. 2001. View Article : Google Scholar : PubMed/NCBI
13.	Sanderson RD and Yang Y: Syndecan-1: a dynamic regulator of the myeloma microenvironment. Clin Exp Metastasis. 25:149–159. 2008. View Article : Google Scholar : PubMed/NCBI
14.	Beauvais DM and Rapraeger AC: Syndecans in tumor cell adhesion and signaling. Reprod Biol Endocrinol. 2:1–12. 2004. View Article : Google Scholar
15.	Tkachenko E, Rhodes JM and Simons M: Syndecans: new kids on the signaling block. Circ Res. 96:488–500. 2005. View Article : Google Scholar : PubMed/NCBI
16.	Fux L, Ilan N, Sanderson RD and Vlodavsky I: Heparanase: busy at the cell surface. Trends Biochem Sci. 34:511–519. 2009. View Article : Google Scholar : PubMed/NCBI
17.	Ilan N, Elkin M and Vlodavsky I: Regulation, function and clinical significance of heparanase in cancer metastasis and angiogenesis. Int J Biochem Cell Biol. 38:2018–2039. 2006. View Article : Google Scholar : PubMed/NCBI
18.	Goodrich LV, Milenkovic L, Higgins KM and Scott MP: Altered neural cell fates and medulloblastoma in mouse patched mutants. Science. 277:1109–1113. 1997. View Article : Google Scholar : PubMed/NCBI
19.	Berman DM, Karhadkar SS, Hallahan AR, et al: Medulloblastoma growth inhibition by hedgehog pathway blockade. Science. 297:1559–1561. 2002. View Article : Google Scholar : PubMed/NCBI
20.	Fattet S, Haberler C, Legoix P, et al: Beta-catenin status in paediatric medulloblastomas: correlation of immunohistochemical expression with mutational status, genetic profiles, and clinical characteristics. J Pathol. 218:86–94. 2009. View Article : Google Scholar
21.	Symons M and Segall JE: Rac and Rho driving tumor invasion: who's at the wheel? Genome Biol. 10:1–4. 2009.PubMed/NCBI
22.	Sanz-Moreno V and Marshall CJ: Rho-GTPase signaling drives melanoma cell plasticity. Cell Cycle. 8:1484–1487. 2009. View Article : Google Scholar : PubMed/NCBI
23.	Ilina O and Friedl P: Mechanisms of collective cell migration at a glance. J Cell Sci. 122:3203–3208. 2009. View Article : Google Scholar : PubMed/NCBI
24.	Avalos AM, Valdivia AD, Munoz N, et al: Neuronal Thy-1 induces astrocyte adhesion by engaging syndecan-4 in a cooperative interaction with alphavbeta3 integrin that activates PKCalpha and RhoA. J Cell Sci. 122:3462–3471. 2009. View Article : Google Scholar : PubMed/NCBI
25.	Sinnappah-Kang ND, Kaiser AJ, Blust BE, Mrak RE and Marchetti D: Heparanase, TrkC and p75NTR: their functional involvement in human medulloblastoma cell invasion. Int J Oncol. 27:617–626. 2005.PubMed/NCBI
26.	Birkenfeld J, Nalbant P, Yoon SH and Bokoch GM: Cellular functions of GEF-H1, a microtubule-regulated Rho-GEF: is altered GEF-H1 activity a crucial determinant of disease pathogenesis? Trends Cell Biol. 18:210–219. 2008. View Article : Google Scholar : PubMed/NCBI
27.	Nardella C, Lahm A, Pallaoro M, Brunetti M, Vannini A and Steinkuhler C: Mechanism of activation of human heparanase investigated by protein engineering. Biochemistry. 43:1862–1873. 2004. View Article : Google Scholar : PubMed/NCBI
28.	Ridgway LD, Kim EY and Dryer SE: MAGI-1 interacts with Slo1 channel proteins and suppresses Slo1 expression on the cell surface. Am J Physiol Cell Physiol. 297:C55–C65. 2009. View Article : Google Scholar : PubMed/NCBI
29.	Xian X, Gopal S and Couchman JR: Syndecans as receptors and organizers of the extracellular matrix. Cell Tissue Res. 339:31–46. 2009. View Article : Google Scholar : PubMed/NCBI
30.	Alexander CM, Reichsman F, Hinkes MT, et al: Syndecan-1 is required for Wnt-1-induced mammary tumorigenesis in mice. Nat Genet. 25:329–332. 2000. View Article : Google Scholar : PubMed/NCBI
31.	Dovas A, Yoneda A and Couchman JR: PKCbeta-dependent activation of RhoA by syndecan-4 during focal adhesion formation. J Cell Sci. 119:2837–2846. 2006. View Article : Google Scholar : PubMed/NCBI
32.	Nalbant P, Chang YC, Birkenfeld J, Chang ZF and Bokoch GM: Guanine nucleotide exchange factor-H1 regulates cell migration via localized activation of RhoA at the leading edge. Mol Biol Cell. 20:4070–4082. 2009. View Article : Google Scholar : PubMed/NCBI
33.	Hartsock A and Nelson WJ: Adherens and tight junctions: structure, function and connections to the actin cytoskeleton. Biochim Biophys Acta. 1778:660–669. 2008. View Article : Google Scholar : PubMed/NCBI
34.	Alvarez-Medina R, Cayuso J, Okubo T, Takada S and Marti E: Wnt canonical pathway restricts graded Shh/Gli patterning activity through the regulation of Gli3 expression. Development. 135:237–247. 2008. View Article : Google Scholar : PubMed/NCBI
35.	Ruiz i Altaba A, Mas C and Stecca B: The Gli code: an information nexus regulating cell fate, stemness and cancer. Trends Cell Biol. 17:438–447. 2007.PubMed/NCBI
36.	Ridley AJ and Hall A: The small GTP-binding protein rho regulates the assembly of focal adhesions and actin stress fibers in response to growth factors. Cell. 70:389–399. 1992. View Article : Google Scholar : PubMed/NCBI
37.	Ridley AJ, Paterson HF, Johnston CL, Diekmann D and Hall A: The small GTP-binding protein rac regulates growth factor-induced membrane ruffling. Cell. 70:401–410. 1992. View Article : Google Scholar : PubMed/NCBI
38.	Sanz-Moreno V, Gadea G, Ahn J, et al: Rac activation and inactivation control plasticity of tumor cell movement. Cell. 135:510–523. 2008. View Article : Google Scholar : PubMed/NCBI
39.	Ren Y, Li R, Zheng Y and Busch H: Cloning and characterization of GEF-H1, a microtubule-associated guanine nucleotide exchange factor for Rac and Rho GTPases. J Biol Chem. 273:34954–34960. 1998. View Article : Google Scholar : PubMed/NCBI
40.	Birkenfeld J, Nalbant P, Bohl BP, Pertz O, Hahn KM and Bokoch GM: GEF-H1 modulates localized RhoA activation during cytokinesis under the control of mitotic kinases. Dev Cell. 12:699–712. 2007. View Article : Google Scholar : PubMed/NCBI
41.	Ulloa F and Marti E: Wnt won the war: antagonistic role of Wnt over Shh controls dorso-ventral patterning of the vertebrate neural tube. Dev Dyn. 239:69–76. 2009.PubMed/NCBI
42.	Sasai K, Romer JT, Lee Y, et al: Shh pathway activity is down-regulated in cultured medulloblastoma cells: implications for preclinical studies. Cancer Res. 66:4215–4222. 2006. View Article : Google Scholar : PubMed/NCBI
43.	Ridgway LD, Wetzel MD and Marchetti D: Modulation of GEF-H1 induced signaling by heparanase in brain metastatic melanoma cells. J Cell Biochem. Aug;2010.(E-pub ahead of print).