Inhibiting the Recruitment of PLCγ1 to Kaposi’s Sarcoma Herpesvirus K15 Protein Reduces the Invasiveness and Angiogenesis of Infected Endothelial Cells

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Kaposi’s Sarcoma (KS), etiologically linked to Kaposi’s sarcoma herpesvirus (KSHV), is a tumour of endothelial origin characterised by angiogenesis and invasiveness. In vitro, KSHV infected endothelial cells display an increased invasiveness and high angiogenicity. Here we report that the KSHV protein K15, which increases the angiogenicity of endothelial cells, contributes to KSHV-mediated invasiveness by the recruitment and activation of the cellular protein PLCγ1 and its downstream effectors βPIX, GIT1 and cdc42. We explored the functional consequences of disrupting the K15-PLCγ1 interaction by using an isolated PLCγ2 cSH2 domain as a dominant negative inhibitor. This protein fragment, by interacting with K15, reduces K15-driven recruitment and activation of PLCγ1 in a dose-dependent manner. Moreover, the PCLγ2 cSH2 domain, when overexpressed in KSHV infected endothelial cells, reduces the angiogenesis and invasiveness induced by the virus. These findings highlight the role of the K15-PLCγ1 interaction in KSHV-mediated invasiveness and identify it as a possible therapeutic target.

Vyšlo v časopise: Inhibiting the Recruitment of PLCγ1 to Kaposi’s Sarcoma Herpesvirus K15 Protein Reduces the Invasiveness and Angiogenesis of Infected Endothelial Cells. PLoS Pathog 11(8): e32767. doi:10.1371/journal.ppat.1005105
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1005105

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1. Chang Y, Cesarman E, Pessin MS, Lee F, Culpepper J, et al. (1994) Identification of herpesvirus-like DNA sequences in AIDS-associated Kaposi's sarcoma. Science 266: 1865–1869. 7997879

2. Chang Y, Moore P (2014) Twenty years of KSHV. Viruses 6: 4258–4264. doi: 10.3390/v6114258 25386844

3. Cesarman E, Chang Y, Moore PS, Said JW, Knowles DM (1995) Kaposi's sarcoma-associated herpesvirus-like DNA sequences in AIDS-related body-cavity-based lymphomas. N Engl J Med 332: 1186–1191. 7700311

4. Soulier J, Grollet L, Oksenhendler E, Cacoub P, Cazals-Hatem D, et al. (1995) Kaposi's sarcoma-associated herpesvirus-like DNA sequences in multicentric Castleman's disease. Blood 86: 1276–1280. 7632932

5. Parkin DM, Bray F, Ferlay J, Jemal A (2014) Cancer in Africa 2012. Cancer Epidemiol Biomarkers Prev 23: 953–966. doi: 10.1158/1055-9965.EPI-14-0281 24700176

6. Rios A (2014) HIV-Related Hematological Malignancies: A Concise Review. Clin Lymphoma Myeloma Leuk 14S: S96–S103.

7. Orenstein JM (2008) Ultrastructure of Kaposi sarcoma. Ultrastruct Pathol 32: 211–220. doi: 10.1080/01913120802343871 18958795

8. Ensoli B, Sturzl M (1998) Kaposi's sarcoma: a result of the interplay among inflammatory cytokines, angiogenic factors and viral agents. Cytokine Growth Factor Rev 9: 63–83. 9720757

9. Ojala PM, Schulz TF (2014) Manipulation of endothelial cells by KSHV: implications for angiogenesis and aberrant vascular differentiation. Semin Cancer Biol 26: 69–77. doi: 10.1016/j.semcancer.2014.01.008 24486643

10. Staskus KA, Zhong W, Gebhard K, Herndier B, Wang H, et al. (1997) Kaposi's sarcoma-associated herpesvirus gene expression in endothelial (spindle) tumor cells. J Virol 71: 715–719. 8985403

11. Ensoli B, Sturzl M, Monini P (2000) Cytokine-mediated growth promotion of Kaposi's sarcoma and primary effusion lymphoma. Semin Cancer Biol 10: 367–381. 11100885

12. Haas DA, Bala K, Busche G, Weidner-Glunde M, Santag S, et al. (2013) The inflammatory kinase MAP4K4 promotes reactivation of Kaposi's sarcoma herpesvirus and enhances the invasiveness of infected endothelial cells. PLoS Pathog 9: e1003737. doi: 10.1371/journal.ppat.1003737 24244164

13. Rosano L, Spinella F, Di Castro V, Nicotra MR, Albini A, et al. (2003) Endothelin receptor blockade inhibits molecular effectors of Kaposi's sarcoma cell invasion and tumor growth in vivo. Am J Pathol 163: 753–762. 12875994

14. Douglas JL, Gustin JK, Dezube B, Pantanowitz JL, Moses AV (2007) Kaposi's sarcoma: a model of both malignancy and chronic inflammation. Panminerva Med 49: 119–138. 17912148

15. Qian LW, Xie J, Ye F, Gao SJ (2007) Kaposi's sarcoma-associated herpesvirus infection promotes invasion of primary human umbilical vein endothelial cells by inducing matrix metalloproteinases. J Virol 81: 7001–7010. 17442715

16. Douglas JL, Gustin JK, Moses AV, Dezube BJ, Pantanowitz L (2010) Kaposi Sarcoma Pathogenesis: A Triad of Viral Infection, Oncogenesis and Chronic Inflammation. Transl Biomed 1. 23082307

17. Bala K, Bosco R, Gramolelli S, Haas DA, Kati S, et al. (2012) Kaposi's sarcoma herpesvirus K15 protein contributes to virus-induced angiogenesis by recruiting PLCgamma1 and activating NFAT1-dependent RCAN1 expression. PLoS Pathog 8: e1002927. doi: 10.1371/journal.ppat.1002927 23028325

18. Choi JK, Lee BS, Shim SN, Li M, Jung JU (2000) Identification of the novel K15 gene at the rightmost end of the Kaposi's sarcoma-associated herpesvirus genome. J Virol 74: 436–446. 10590133

19. Glenn M, Rainbow L, Aurade F, Davison A, Schulz TF (1999) Identification of a spliced gene from Kaposi's sarcoma-associated herpesvirus encoding a protein with similarities to latent membrane proteins 1 and 2A of Epstein-Barr virus. J Virol 73: 6953–6963. 10400794

20. Poole LJ, Zong JC, Ciufo DM, Alcendor DJ, Cannon JS, et al. (1999) Comparison of genetic variability at multiple loci across the genomes of the major subtypes of Kaposi's sarcoma-associated herpesvirus reveals evidence for recombination and for two distinct types of open reading frame K15 alleles at the right-hand end. J Virol 73: 6646–6660. 10400762

21. Kakoola DN, Sheldon J, Byabazaire N, Bowden RJ, Katongole-Mbidde E, et al. (2001) Recombination in human herpesvirus-8 strains from Uganda and evolution of the K15 gene. J Gen Virol 82: 2393–2404. 11562533

22. Brinkmann MM, Glenn M, Rainbow L, Kieser A, Henke-Gendo C, et al. (2003) Activation of mitogen-activated protein kinase and NF-kappaB pathways by a Kaposi's sarcoma-associated herpesvirus K15 membrane protein. J Virol 77: 9346–9358. 12915550

23. Havemeier A, Gramolelli S, Pietrek M, Jochmann R, Sturzl M, et al. (2014) Activation of NF-kappaB by the Kaposi's sarcoma-associated herpesvirus K15 protein involves recruitment of the NF-kappaB-inducing kinase, IkappaB kinases, and phosphorylation of p65. J Virol 88: 13161–13172. doi: 10.1128/JVI.01766-14 25187543

24. Wang L, Pietrek M, Brinkmann MM, Havemeier A, Fischer I, et al. (2009) Identification and functional characterization of a spliced rhesus rhadinovirus gene with homology to the K15 gene of Kaposi's sarcoma-associated herpesvirus. J Gen Virol 90: 1190–1201. doi: 10.1099/vir.0.007971-0 19264656

25. Brinkmann MM, Pietrek M, Dittrich-Breiholz O, Kracht M, Schulz TF (2007) Modulation of host gene expression by the K15 protein of Kaposi's sarcoma-associated herpesvirus. J Virol 81: 42–58. 17050609

26. Sala G, Dituri F, Raimondi C, Previdi S, Maffucci T, et al. (2008) Phospholipase Cgamma1 is required for metastasis development and progression. Cancer Res 68: 10187–10196. doi: 10.1158/0008-5472.CAN-08-1181 19074886

27. Jones NP, Katan M (2007) Role of phospholipase Cgamma1 in cell spreading requires association with a beta-Pix/GIT1-containing complex, leading to activation of Cdc42 and Rac1. Mol Cell Biol 27: 5790–5805. 17562871

28. Jones NP, Peak J, Brader S, Eccles SA, Katan M (2005) PLCgamma1 is essential for early events in integrin signalling required for cell motility. J Cell Sci 118: 2695–2706. 15944397

29. Kassis J, Lauffenburger DA, Turner T, Wells A (2001) Tumor invasion as dysregulated cell motility. Semin Cancer Biol 11: 105–117. 11322830

30. Kassis J, Moellinger J, Lo H, Greenberg NM, Kim HG, et al. (1999) A role for phospholipase C-gamma-mediated signaling in tumor cell invasion. Clin Cancer Res 5: 2251–2260. 10473113

31. Lattanzio R, Marchisio M, La Sorda R, Tinari N, Falasca M, et al. (2013) Overexpression of activated phospholipase Cgamma1 is a risk factor for distant metastases in T1–T2, N0 breast cancer patients undergoing adjuvant chemotherapy. Int J Cancer 132: 1022–1031. doi: 10.1002/ijc.27751 22847294

32. Lattanzio R, Piantelli M, Falasca M (2013) Role of phospholipase C in cell invasion and metastasis. Adv Biol Regul 53: 309–318. doi: 10.1016/j.jbior.2013.07.006 23925006

33. Kunze K, Spieker T, Gamerdinger U, Nau K, Berger J, et al. (2014) A recurrent activating PLCG1 mutation in cardiac angiosarcomas increases apoptosis resistance and invasiveness of endothelial cells. Cancer Res 74: 6173–6183. doi: 10.1158/0008-5472.CAN-14-1162 25252913

34. Behjati S, Tarpey PS, Sheldon H, Martincorena I, Van Loo P, et al. (2014) Recurrent PTPRB and PLCG1 mutations in angiosarcoma. Nat Genet 46: 376–379. doi: 10.1038/ng.2921 24633157

35. Koss H, Bunney TD, Behjati S, Katan M (2014) Dysfunction of phospholipase Cgamma in immune disorders and cancer. Trends Biochem Sci 39: 603–611. doi: 10.1016/j.tibs.2014.09.004 25456276

36. Sharp TV, Wang HW, Koumi A, Hollyman D, Endo Y, et al. (2002) K15 protein of Kaposi's sarcoma-associated herpesvirus is latently expressed and binds to HAX-1, a protein with antiapoptotic function. J Virol 76: 802–816. 11752170

37. Tsai YH, Wu MF, Wu YH, Chang SJ, Lin SF, et al. (2009) The M type K15 protein of Kaposi's sarcoma-associated herpesvirus regulates microRNA expression via its SH2-binding motif to induce cell migration and invasion. J Virol 83: 622–632. doi: 10.1128/JVI.00869-08 18971265

38. Manabe R, Kovalenko M, Webb DJ, Horwitz AR (2002) GIT1 functions in a motile, multi-molecular signaling complex that regulates protrusive activity and cell migration. J Cell Sci 115: 1497–1510. 11896197

39. Chattopadhyay A, Vecchi M, Ji Q, Mernaugh R, Carpenter G (1999) The role of individual SH2 domains in mediating association of phospholipase C-gamma1 with the activated EGF receptor. J Biol Chem 274: 26091–26097. 10473558

40. Katan M, Williams RL (1997) Phosphoinositide-specific phospholipase C: structural basis for catalysis and regulatory interactions. Semin Cell Dev Biol 8: 287–296. 10024492

41. Scott JD, Pawson T (2009) Cell signaling in space and time: where proteins come together and when they're apart. Science 326: 1220–1224. doi: 10.1126/science.1175668 19965465

42. Liu BA, Engelmann BW, Nash PD (2012) The language of SH2 domain interactions defines phosphotyrosine-mediated signal transduction. FEBS Lett 586: 2597–2605. doi: 10.1016/j.febslet.2012.04.054 22569091

43. Liu BA, Jablonowski K, Shah EE, Engelmann BW, Jones RB, et al. (2010) SH2 domains recognize contextual peptide sequence information to determine selectivity. Mol Cell Proteomics 9: 2391–2404. doi: 10.1074/mcp.M110.001586 20627867

44. Ji QS, Winnier GE, Niswender KD, Horstman D, Wisdom R, et al. (1997) Essential role of the tyrosine kinase substrate phospholipase C-gamma1 in mammalian growth and development. Proc Natl Acad Sci U S A 94: 2999–3003. 9096335

45. Decker C, Hesker P, Zhang K, Faccio R (2013) Targeted inhibition of phospholipase C gamma2 adaptor function blocks osteoclastogenesis and protects from pathological osteolysis. J Biol Chem 288: 33634–33641. doi: 10.1074/jbc.M113.477281 24081142

46. Kaneko T, Huang H, Cao X, Li X, Li C, et al. (2012) Superbinder SH2 domains act as antagonists of cell signaling. Sci Signal 5: ra68. 23012655

47. DeBell K, Graham L, Reischl I, Serrano C, Bonvini E, et al. (2007) Intramolecular regulation of phospholipase C-gamma1 by its C-terminal Src homology 2 domain. Mol Cell Biol 27: 854–863. 17116690

48. Gresset A, Hicks SN, Harden TK, Sondek J (2010) Mechanism of phosphorylation-induced activation of phospholipase C-gamma isozymes. J Biol Chem 285: 35836–35847. doi: 10.1074/jbc.M110.166512 20807769

49. Gramolelli S, Schulz TF (2015) The role of Kaposi sarcoma-associated herpesvirus in the pathogenesis of Kaposi sarcoma. J Pathol 235: 368–380. doi: 10.1002/path.4441 25212381

50. Frank SR, Hansen SH (2008) The PIX-GIT complex: a G protein signaling cassette in control of cell shape. Semin Cell Dev Biol 19: 234–244. doi: 10.1016/j.semcdb.2008.01.002 18299239

51. Hoefen RJ, Berk BC (2006) The multifunctional GIT family of proteins. J Cell Sci 119: 1469–1475. 16598076

52. Vieira J, O'Hearn PM (2004) Use of the red fluorescent protein as a marker of Kaposi's sarcoma-associated herpesvirus lytic gene expression. Virology 325: 225–240. 15246263

53. Bunney TD, Esposito D, Mas-Droux C, Lamber E, Baxendale RW, et al. (2012) Structural and functional integration of the PLCgamma interaction domains critical for regulatory mechanisms and signaling deregulation. Structure 20: 2062–2075. doi: 10.1016/j.str.2012.09.005 23063561

54. Chen P, Xie H, Sekar MC, Gupta K, Wells A (1994) Epidermal growth factor receptor-mediated cell motility: phospholipase C activity is required, but mitogen-activated protein kinase activity is not sufficient for induced cell movement. J Cell Biol 127: 847–857. 7962064

55. Wang L, Damania B (2008) Kaposi's sarcoma-associated herpesvirus confers a survival advantage to endothelial cells. Cancer Res 68: 4640–4648. doi: 10.1158/0008-5472.CAN-07-5988 18559509

56. Wang L, Brinkmann MM, Pietrek M, Ottinger M, Dittrich-Breiholz O, et al. (2007) Functional characterization of the M-type K15-encoded membrane protein of Kaposi's sarcoma-associated herpesvirus. J Gen Virol 88: 1698–1707. 17485529

57. Jaffe EA, Nachman RL, Becker CG, Minick CR (1973) Culture of human endothelial cells derived from umbilical veins. Identification by morphologic and immunologic criteria. J Clin Invest 52: 2745–2756. 4355998

58. May T, Butueva M, Bantner S, Markusic D, Seppen J, et al. (2010) Synthetic gene regulation circuits for control of cell expansion. Tissue Eng Part A 16: 441–452. doi: 10.1089/ten.TEA.2009.0184 19705962

59. Alkharsah KR, Singh VV, Bosco R, Santag S, Grundhoff A, et al. (2011) Deletion of Kaposi's sarcoma-associated herpesvirus FLICE inhibitory protein, vFLIP, from the viral genome compromises the activation of STAT1-responsive cellular genes and spindle cell formation in endothelial cells. J Virol 85: 10375–10388. doi: 10.1128/JVI.00226-11 21795355

60. Kati S, Hage E, Mynarek M, Ganzenmueller T, Indenbirken D, et al. (2015) Generation of high-titre virus stocks using BrK.219, a B-cell line infected stably with recombinant Kaposi's sarcoma-associated herpesvirus. J Virol Methods.

61. Carpenter AE, Jones TR, Lamprecht MR, Clarke C, Kang IH, et al. (2006) CellProfiler: image analysis software for identifying and quantifying cell phenotypes. Genome Biol 7: R100. 17076895