Identification of BIRC6 as a novel intervention target for neuroblastoma therapy

BMC Cancer

Fieke Lamers²  Linda Schild²  Jan Koster²  Frank Speleman³  Ingrid Øra⁵  Ellen M Westerhout²  Peter van Sluis²  Rogier Versteeg²  Huib N Caron¹  Jan J Molenaar⁴ 

[1] Department of Pediatric Oncology, Emma Kinderziekenhuis, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands

[2] Department of Oncogenomics, Academic Medical Center, University of Amsterdam, Meibergdreef 15, PO box 22700, Amsterdam, AZ 1105, The Netherlands

[3] Center for Medical Genetics, Ghent University Hospital, Ghent, Belgium

[4] Department of Oncogenomics, M1-132, Academic Medical Center, University of Amsterdam, Meibergdreef 9, Amsterdam, AZ 1105, The Netherlands

[5] Department of Pediatric Oncology, Skåne University Hospital, Lund University, Lund, Sweden

关键词: Cancer;    Apoptosis;    DIABLO;    BIRC6;    Neuroblastoma;   

Others  :  1080322
DOI  :  10.1186/1471-2407-12-285

 received in 2012-02-27, accepted in 2012-06-26,  发布年份 2012

【 摘 要 】

Background

Neuroblastoma are pediatric tumors of the sympathetic nervous system with a poor prognosis. Apoptosis is often deregulated in cancer cells, but only a few defects in apoptotic routes have been identified in neuroblastoma.

Methods

Here we investigated genomic aberrations affecting genes of the intrinsic apoptotic pathway in neuroblastoma. We analyzed DNA profiling data (CGH and SNP arrays) and mRNA expression data of 31 genes of the intrinsic apoptotic pathway in a dataset of 88 neuroblastoma tumors using the R2 bioinformatic platform ( http://r2.amc.nl webcite). BIRC6 was selected for further analysis as a tumor driving gene. Knockdown experiments were performed using BIRC6 lentiviral shRNA and phenotype responses were analyzed by Western blot and MTT-assays. In addition, DIABLO levels and interactions were investigated with immunofluorescence and co-immunoprecipitation.

Results

We observed frequent gain of the BIRC6 gene on chromosome 2, which resulted in increased mRNA expression. BIRC6 is an inhibitor of apoptosis protein (IAP), that can bind and degrade the cytoplasmic fraction of the pro-apoptotic protein DIABLO. DIABLO mRNA expression was exceptionally high in neuroblastoma but the protein was only detected in the mitochondria. Upon silencing of BIRC6 by shRNA, DIABLO protein levels increased and cells went into apoptosis. Co-immunoprecipitation confirmed direct interaction between DIABLO and BIRC6 in neuroblastoma cell lines.

Conclusion

Our findings indicate that BIRC6 may have a potential oncogenic role in neuroblastoma by inactivating cytoplasmic DIABLO. BIRC6 inhibition may therefore provide a means for therapeutic intervention in neuroblastoma.

【 授权许可】

   
2012 Lamers et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

【 参考文献 】

• [1]Hao YY, Sekine K, Kawabata A, Nakamura H, Ishioka T, Ohata H, Katayama R, Hashimoto C, Zhang XD, Noda T, et al.: Apollon ubiquitinates SMAC and caspase-9, and has an essential cytoprotection function. Nat Cell Biol 2004, 6:849-860.
• [2]Qiu XB, Goldberg AL: The membrane-associated inhibitor of apoptosis protein, BRUCE/Apollon, antagonizes both the precursor and mature forms of Smac and caspase-9. J Biol Chem 2005, 280:174-182.
• [3]Buron N, Porceddu M, Brabant M, Desgue D, Racoeur C, Lassalle M, Pechoux C, Rustin P, Jacotot E, Borgne-Sanchez A: Use of human cancer cell lines mitochondria to explore the mechanisms of BH3 Peptides and ABT-737-induced mitochondrial membrane permeabilization. PLoS One 2010, 5(3):e9924.
• [4]Ceballos-Cancino G, Espinosa M, Maldonado V, Melendez-Zajgla J: Regulation of mitochondrial Smac/DIABLO-selective release by survivin. Oncogene 2007, 26:7569-7575.
• [5]Yu J, Wang P, Ming L, Wood MA, Zhang L: SMAC/Diablo mediates the proapoptotic function of PUMA by regulating PUMA-induced mitochondrial events. Oncogene 2007, 26:4189-4198.
• [6]Du CY, Fang M, Li YC, Li L, Wang XD: Smac, a mitochondrial protein that promotes cytochrome c-dependent caspase activation by eliminating IAP inhibition. Cell 2000, 102:33-42.
• [7]Martin SJ: An Apollon vista of death and destruction. Nat Cell Biol 2004, 6:804-806.
• [8]Pohl C, Jentsch S: Final stages of cytokinesis and midbody ring formation are controlled by BRUCE. Cell 2008, 132:832-845.
• [9]Sung KW, Choi J, Hwang YK, Lee SJ, Kim HJ, Lee SH, HeeYbo K, Jung HL, Koo HH: Overexpression of apollon, an antiapoptotic protein, is associated with poor prognosis in childhood De novo acute myeloid leukemia. Clin Cancer Res 2007, 13:5109-5114.
• [10]Toujani S, Dessen P, Ithzar N, Danglot G, Richon C, Vassetzky Y, Robert T, Lazar V, Bosq J, Da Costa L, et al.: High resolution genome-wide analysis of chromosomal alterations in Burkitt's Lymphoma. PLoS One 2009, 4(9):e7089.
• [11]Van Houdt WJ, Emmink BL, Pham TV, Piersma SR, Verheem A, Vries RG, Fratantoni SA, Pronk A, Clevers H, Rinkes IHMB, et al.: Comparative proteomics of colon cancer stem cells and differentiated tumor cells identifies BIRC6 as a potential therapeutic target. Mol Cell Proteomics 2011, 10(12):M111.011353.
• [12]Brodeur GM: Neuroblastoma: biological insights into a clinical enigma. Nat Rev Cancer 2003, 3:203-216.
• [13]Maris JM, Hogarty MD, Bagatell R, Cohn SL: Neuroblastoma. Lancet 2007, 369:2106-2120.
• [14]Maris JM: Medical progress: recent advances in neuroblastoma. N Engl J Med 2010, 362:2202-2211.
• [15]Meyer N, Penn LZ: MYC - TIMELINE Reflecting on 25 years with MYC. Nat Rev Cancer 2008, 8:976-990.
• [16]Chen YY, Takita J, Choi YL, Kato M, Ohira M, Sanada M, Wang LL, Soda M, Kikuchi A, Igarashi T, et al.: Oncogenic mutations of ALK kinase in neuroblastoma. Nature 2008, 455:971-974.
• [17]George RE, Sanda T, Hanna M, Frohling S, Luther W, Zhang JM, Ahn Y, Zhou WJ, London WB, McGrady P, et al.: Activating mutations in ALK provide a therapeutic target in neuroblastoma. Nature 2008, 455:975-978.
• [18]Janoueix-Lerosey I, Lequin D, Brugieres L, Ribeiro A, de Pontual L, Combaret V, Raynal V, Puisieux A, Schleiermacher G, Pierron G, et al.: Somatic and germline activating mutations of the ALK kinase receptor in neuroblastoma. Nature 2008, 455:967-970.
• [19]Janoueix-Lerosey I, Schleiermacher G, Delattre O: Molecular pathogenesis of peripheral neuroblastic tumors. Oncogene 2010, 29:1566-1579.
• [20]Mosse YP, Laudenslager M, Longo L, Cole KA, Wood A, Attiyeh EF, Laquaglia MJ, Sennett R, Lynch JE, Perri P, et al.: Identification of ALK as a major familial neuroblastoma predisposition gene. Nature 2008, 455:930-935.
• [21]De Brouwer S, De Preter K, Kumps C, Zabrocki P, Porcu M, Westerhout EM, Lakeman A, Vandesompele J, Hoebeeck J, Van Maerken T, et al.: Meta-analysis of Neuroblastomas Reveals a Skewed ALK Mutation Spectrum in Tumors with MYCN Amplification. Clin Cancer Res 2010, 16:4353-4362.
• [22]Van Maerken T, Ferdinande L, Taildeman J, Lambertz I, Yigit N, Vercruysse L, Rihani A, Michaelis M, Cinatl J, Cuvelier CA, et al.: Antitumor activity of the selective MDM2 antagonist nutlin-3 against chemoresistant neuroblastoma with wild-type p53. J Natl Cancer Inst 2009, 101:1562-1574.
• [23]van Noesel MM, Versteeg R: Pediatric neuroblastomas: genetic and epigenetic 'Danse Macabre'. Gene 2004, 325:1-15.
• [24]Adida C, Berrebi D, Peuchmaur M, Reyes-Mugica M, Altieri DC: Anti-apoptosis gene, survivin, and prognosis of neuroblastoma. Lancet 1998, 351:882-883.
• [25]Islam A, Kageyama H, Takada N, Kawamato T, Takayasu H, Isogai E, Ohira M, Hashizume K, Kobayashi H, Kaneko Y, et al.: High expression of Survivin, mapped to 17q25, is significantly associated with poor prognostic factors and promotes cell survival in human neuroblastoma. Oncogene 2000, 19:617-623.
• [26]Lamers F, van der Ploeg I, Schild L, Ebus ME, Koster J, Hansen BR, Koch T, Versteeg R, Caron HN, Molenaar JJ: Knockdown of Survivin (BIRC5) causes Apoptosis in Neuroblastoma via Mitotic Catastrophe. Endocr Relat Cancer 2011, 18:657-668.
• [27]Giaccone G, Zatloukal P, Roubec J, Floor K, Musil J, Kuta M, van Klaveren RJ, Chaudhary S, Gunther A, Shamsili S: Multicenter Phase II Trial of YM155, a small-molecule suppressor of survivin, in patients with advanced, refractory, non-small-cell lung cancer. J Clin Oncol 2009, 27:4481-4486.
• [28]Lamers F, Schild L, Koster J, Versteeg R, Caron H, Molenaar J: Targeted BIRC5 silencing using YM155 causes cell death in neuroblastoma cells with low ABCB1 expression. Eur J Cancer 2012, 48(5):763-771.
• [29]Lewis KD, Samlowski W, Ward J, Catlett J, Cranmer L, Kirkwood J, Lawson D, Whitman E, Gonzalez R: A multi-center phase II evaluation of the small molecule survivin suppressor YM155 in patients with unresectable stage III or IV melanoma. Invest New Drugs 2011, 29(1):161-166.
• [30]Nakahara T, Takeuchi M, Kinoyama I, Minematsu T, Shirasuna K, Matsuhisa A, Kita A, Tominaga F, Yamanaka K, Kudoh M, et al.: YM155, a novel small-molecule survivin suppressant, induces regression of established human hormone-refractory prostate tumor xenografts. Cancer Res 2007, 67:8014-8021.
• [31]Satoh T, Okamoto I, Miyazaki M, Morinaga R, Tsuya A, Hasegawa Y, Terashima M, Ueda S, Fukuoka M, Ariyoshi Y, et al.: Phase I Study of YM155, a novel survivin suppressant, in patients with advanced solid tumors. Clin Cancer Res 2009, 15:3872-3880.
• [32]Tolcher AW, Mita A, Lewis LD, Garrett CR, Till E, Daud AI, Patnaik A, Papadopoulos K, Takimoto C, Bartels P, et al.: Phase I and Pharmacokinetic Study of YM155, a Small-Molecule Inhibitor of Survivin. J Clin Oncol 2008, 26:5198-5203.
• [33]Lamers F, Schild L, den Hartog IJM, Ebus ME, Westerhout EM, Ora I, Koster J, Versteeg R, Caron HN, Molenaar JJ: Targeted BCL2 inhibition effectively inhibits Neuroblastoma tumor growth. Eur J Cancerin press
• [34]Santo EE, Ebus ME, Koster J, Schulte JH, Lakeman A, van Sluis P, Vermeulen J, Gisselsson D, Ora I, Lindner S, et al.: Oncogenic activation of FOXR1 by 11q23 intrachromosomal deletion-fusions in neuroblastoma. Oncogene 2012, 31(12):1571-1581.
• [35]Molenaar JJ, Ebus ME, Koster J, van Sluis P, van Noesel CJM, Versteeg R, Caron HN: Cyclin D1 and CDK4 activity contribute to the undifferentiated phenotype in neuroblastoma. Cancer Res 2008, 68:2599-2609.
• [36]Fix A, Lucchesi C, Ribeiro A, Lequin D, Pierron G, Schleiermacher G, Delattre O, Janoueix-Lerosey I: Characterization of amplicons in neuroblastoma: high-resolution mapping using DNA microarrays, relationship with outcome, and identification of overexpressed genes. Genes Chromosomes Cancer 2008, 47:819-834.
• [37]Mullenbach R, Lagoda PJL, Welter C: An efficient salt-chloroform extraction of dna from blood and tissues. Trends Genet 1989, 5:391.
• [38]Ren JY, Shi MG, Liu RS, Yang QH, Johnson T, Skarnes WC, Du CY: The Birc6 (Bruce) gene regulates p53 and the mitochondrial pathway of apoptosis and is essential for mouse embryonic development. Proc Natl Acad Sci USA 2005, 102:565-570.
• [39]Basu A, Adkins B, Basu C: Down-regulation of caspase-2 by rottlerin via protein kinase c-delta-independent pathway. Cancer Res 2008, 68:2795-2802.
• [40]Nutt LK, Margolis SS, Jensen M, Herman CE, Dunphy WG, Rathmell JC, Kornbluth S: Metabolic regulation of oocyte cell death through the CaMKII-mediated phosphorylation of caspase-2. Cell 2005, 123:89-103.
• [41]Shen J, Vakifahmetoglu H, Stridh H, Zhivotovsky B, Wiman KG: PRIMA-1(MET) induces mitochondrial apoptosis through activation of caspase-2. Oncogene 2008, 27:6571-6580.
• [42]Wolf M, Korja M, Karhu R, Edgren H, Kilpinen S, Ojala K, Mousses S, Kallioniemi A, Haapasalo H: Array-based gene expression, CGH and tissue data defines a 12q24 gain in neuroblastic tumors with prognostic implication. BMC Cancer 2010, 10:181. BioMed Central Full Text
• [43]Lestini BJ, Goldsmith KC, Fluchel MN, Liu XY, Chen NL, Goyal B, Pawel BR, Hogarty MD: Mcl1 downregulation sensitizes neuroblastoma to cytotoxic chemotherapy and small molecule Bcl2-family antagonists. Cancer Biol Ther 2009, 8:1587-1595.
• [44]Kushnareva Y, Newmeyer DD: Bioenergetics and cell death. Mitochondrial Res Transl Med 2010, 1201:50-57.
• [45]Ekert PG, Silke J, Hawkins CJ, Verhagen AM, Vaux DL: DIABLO promotes apoptosis by removing MIHA/XIAP from processed caspase 9. J Cell Biol 2001, 152:483-490.
• [46]Teitz T, Wei T, Liu D, Valentine V, Valentine M, Grenet J, Lahti JM, Kidd VJ: Caspase-9 and Apaf-1 are expressed and functionally active in human neuroblastoma tumor cell lines with 1p36 LOH and amplified MYCN. Oncogene 2002, 21:1848-1858.