, a -Antisense Gene of , Encodes a Evolved Protein That Inhibits GSK3β Resulting in the Stabilization of MYCN in Human Neuroblastomas

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The rearrangement of pre-existing genes has long been thought of as the major mode of new gene generation. Recently, de novo gene birth from non-genic DNA was found to be an alternative mechanism to generate novel protein-coding genes. However, its functional role in human disease remains largely unknown. Here we show that NCYM, a cis-antisense gene of the MYCN oncogene, initially thought to be a large non-coding RNA, encodes a de novo evolved protein regulating the pathogenesis of human cancers, particularly neuroblastoma. The NCYM gene is evolutionally conserved only in the taxonomic group containing humans and chimpanzees. In primary human neuroblastomas, NCYM is 100% co-amplified and co-expressed with MYCN, and NCYM mRNA expression is associated with poor clinical outcome. MYCN directly transactivates both NCYM and MYCN mRNA, whereas NCYM stabilizes MYCN protein by inhibiting the activity of GSK3β, a kinase that promotes MYCN degradation. In contrast to MYCN transgenic mice, neuroblastomas in MYCN/NCYM double transgenic mice were frequently accompanied by distant metastases, behavior reminiscent of human neuroblastomas with MYCN amplification. The NCYM protein also interacts with GSK3β, thereby stabilizing the MYCN protein in the tumors of the MYCN/NCYM double transgenic mice. Thus, these results suggest that GSK3β inhibition by NCYM stabilizes the MYCN protein both in vitro and in vivo. Furthermore, the survival of MYCN transgenic mice bearing neuroblastoma was improved by treatment with NVP-BEZ235, a dual PI3K/mTOR inhibitor shown to destabilize MYCN via GSK3β activation. In contrast, tumors caused in MYCN/NCYM double transgenic mice showed chemo-resistance to the drug. Collectively, our results show that NCYM is the first de novo evolved protein known to act as an oncopromoting factor in human cancer, and suggest that de novo evolved proteins may functionally characterize human disease.

Vyšlo v časopise: , a -Antisense Gene of , Encodes a Evolved Protein That Inhibits GSK3β Resulting in the Stabilization of MYCN in Human Neuroblastomas. PLoS Genet 10(1): e32767. doi:10.1371/journal.pgen.1003996
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.pgen.1003996

Souhrn

Zdroje

1. JacobF (1977) Evolution and tinkering. Science 196: 1161–1166.

2. Ohno S (1970) Evolution by gene duplication. New York, NY, Springer-Verlag.

3. TautzD, Domazet-LosoT (2011) The evolutionary origin of orphan genes. Nat Rev Genet 12: 692–702.

4. KaessmannH (2010) Origins, evolution, and phenotypic impact of new genes. Genome Res 20: 1313–1326.

5. KhalturinK, HemmrichG, FrauneS, AugustinR, BoschTC (2009) More than just orphans: are taxonomically-restricted genes important in evolution? Trends Genet 25: 404–413.

6. CarvunisAR, RollandT, WapinskiI, CalderwoodMA, YildirimMA, et al. (2012) Proto-genes and de novo gene birth. Nature 487: 370–374.

7. LiD, DongY, JiangY, JiangH, CaiJ, et al. (2010) A de novo originated gene depresses budding yeast mating pathway and is repressed by the protein encoded by its antisense strand. Cell Res 20: 408–420.

8. BegunDJ, LindforsHA, ThompsonME, HollowayAK (2006) Recently evolved genes identified from Drosophila yakuba and D. erecta accessory gland expressed sequence tags. Genetics 172: 1675–1681.

9. BegunDJ, LindforsHA, KernAD, JonesCD (2007) Evidence for de novo evolution of testis-expressed genes in the Drosophila yakuba/Drosophila erecta clade. Genetics 176: 1131–1137.

10. CaiJ, ZhaoR, JiangH, WangW (2008) De novo origination of a new protein-coding gene in Saccharomyces cerevisiae. Genetics 179: 487–496.

11. ChenST, ChengHC, BarbashDA, YangHP (2007) Evolution of hydra, a recently evolved testis-expressed gene with nine alternative first exons in Drosophila melanogaster. PLoS Genet 3: e107.

12. Toll-RieraM, BoschN, BelloraN, CasteloR, ArmengolL, et al. (2009) Origin of primate orphan genes: a comparative genomics approach. Mol Biol Evol 26: 603–612.

13. KnowlesDG, McLysaghtA (2009) Recent de novo origin of human protein-coding genes. Genome Res 19: 1752–1759.

14. LiCY, ZhangY, WangZ, CaoC, ZhangPW, et al. (2010) A human-specific de novo protein-coding gene associated with human brain functions. PLoS Comput Biol 6: e1000734.

15. O'BlenessM, SearlesVB, VarkiA, GagneuxP, SikelaJM (2012) Evolution of genetic and genomic features unique to the human lineage. Nat Rev Genet 13: 853–866.

16. WuDD, IrwinDM, ZhangYP (2011) De novo origin of human protein-coding genes. PLoS Genet 7: e1002379.

17. XieC, ZhangYE, ChenJY, LiuCJ, ZhouWZ, et al. (2012) Hominoid-specific de novo protein-coding genes originating from long non-coding RNAs. PLoS Genet 8: e1002942.

18. BrodeurGM (2003) Neuroblastoma: biological insights into a clinical enigma. Nat Rev Cancer 3: 203–216.

19. BrodeurGM, SeegerRC, SchwabM, VarmusHE, BishopJM (1984) Amplification of N-myc in untreated human neuroblastomas correlates with advanced disease stage. Science 224: 1121–1124.

20. SchwabM, VarmusHE, BishopJM, GrzeschikKH, NaylorSL, et al. (1984) Chromosome localization in normal human cells and neuroblastomas of a gene related to c-myc. Nature 308: 288–291.

21. WeissWA, AldapeK, MohapatraG, FeuersteinBG, BishopJM (1997) Targeted expression of MYCN causes neuroblastoma in transgenic mice. EMBO J 16: 2985–2995.

22. CohnSL, LondonWB, HuangD, KatzensteinHM, SalwenHR, et al. (2000) MYCN expression is not prognostic of adverse outcome in advanced-stage neuroblastoma with nonamplified MYCN. J Clin Oncol 18: 3604–3613.

23. NakagawaraA, ArimaM, AzarCG, ScavardaNJ, BrodeurGM (1992) Inverse relationship between trk expression and N-myc amplification in human neuroblastomas. Cancer Res 52: 1364–1368.

24. ArmstrongBC, KrystalGW (1992) Isolation and characterization of complementary DNA for N-cym, a gene encoded by the DNA strand opposite to N-myc. Cell Growth Differ 3: 385–390.

25. KrystalGW, ArmstrongBC, BatteyJF (1990) N-myc mRNA forms an RNA-RNA duplex with endogenous antisense transcripts. Mol Cell Biol 10: 4180–4191.

26. JacobsJF, van BokhovenH, van LeeuwenFN, Hulsbergen-van de KaaCA, de VriesIJ, et al. (2009) Regulation of MYCN expression in human neuroblastoma cells. BMC Cancer 9: 239.

27. SuenagaY, KanekoY, MatsumotoD, HossainMS, OzakiT, et al. (2009) Positive auto-regulation of MYCN in human neuroblastoma. Biochem Biophys Res Commun 390: 21–26.

28. GustafsonWC, WeissWA (2010) Myc proteins as therapeutic targets. Oncogene 29: 1249–1259.

29. SjostromSK, FinnG, HahnWC, RowitchDH, KenneyAM (2005) The Cdk1 complex plays a prime role in regulating N-myc phosphorylation and turnover in neural precursors. Dev Cell 9: 327–338.

30. KangJH, RychahouPG, IsholaTA, QiaoJ, EversBM, et al. (2006) MYCN silencing induces differentiation and apoptosis in human neuroblastoma cells. Biochem Biophys Res Commun 351: 192–197.

31. ZhangHH, LipovskyAI, DibbleCC, SahinM, ManningBD (2006) S6K1 regulates GSK3 under conditions of mTOR-dependent feedback inhibition of Akt. Mol Cell 24: 185–197.

32. ChantheryYH, GustafsonWC, ItsaraM, PerssonA, HackettCS, et al. (2012) Paracrine signaling through MYCN enhances tumor-vascular interactions in neuroblastoma. Sci Transl Med 4: 115ra113.

33. SchrammA, KösterJ, MarschallT, MartinM, SchwermerM, et al. (2013) Next-generation RNA sequencing reveals differential expression of MYCN target genes and suggests the mTOR pathway as a promising therapy target in MYCN-amplified neuroblastoma. Int J Cancer 132: E106–115.

34. JohnsenJI, SegerströmL, OrregoA, ElfmanL, HenrikssonM, et al. (2008) Inhibitors of mammalian target of rapamycin downregulate MYCN protein expression and inhibit neuroblastoma growth in vitro and in vivo. Oncogene 27: 2910–2922.

35. YuF, GaoW, YokochiT, SuenagaY, AndoK, et al. (2013) RUNX3 interacts with MYCN and facilitates protein degradation in neuroblastoma. Oncogene [epub ahead of print].

36. OttoT, HornS, BrockmannM, EilersU, SchüttrumpfL, et al. (2009) Stabilization of N-Myc is a critical function of Aurora A in human neuroblastoma. Cancer Cell 15: 67–78.

37. BerryT, LutherW, BhatnagarN, JaminY, PoonE, et al. (2012) The ALK(F1174L) mutation potentiates the oncogenic activity of MYCN in neuroblastoma. Cancer Cell 22: 117–130.

38. HeukampLC, ThorT, SchrammA, De PreterK, KumpsC, et al. (2012) Targeted expression of mutated ALK induces neuroblastoma in transgenic mice. Sci Transl Med 4: 141ra191.

39. MolenaarJJ, Domingo-FernandezR, EbusME, LindnerS, KosterJ, et al. (2012) LIN28B induces neuroblastoma and enhances MYCN levels via let-7 suppression. Nat Genet 44: 1199–1206.

40. ValentijnLJ, KosterJ, HaneveldF, AissaRA, van SluisP, et al. (2012) Functional MYCN signature predicts outcome of neuroblastoma irrespective of MYCN amplification. Proc Natl Acad Sci U S A 109: 19190–19195.

41. LiZ, TanF, ThieleCJ (2007) Inactivation of glycogen synthase kinase-3beta contributes to brain-derived neutrophic factor/TrkB-induced resistance to chemotherapy in neuroblastoma cells. Mol Cancer Ther 6: 3113–3121.

42. KentWJ (2002) BLAT–the BLAST-like alignment tool. Genome Res 12: 656–664.

43. BlanchetteM, KentWJ, RiemerC, ElnitskiL, SmitAF, et al. (2004) Aligning multiple genomic sequences with the threaded blockset aligner. Genome Res 14: 708–715.

44. AltschulSF, MaddenTL, SchafferAA, ZhangJ, ZhangZ, et al. (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 25: 3389–3402.

45. WangD, ZhangY, ZhangZ, ZhuJ, YuJ (2010) KaKs_Calculator 2.0: a toolkit incorporating gamma-series methods and sliding window strategies. Genomics Proteomics Bioinformatics 8: 77–80.

46. OhiraM, ObaS, NakamuraY, IsogaiE, KanekoS, et al. (2005) Expression profiling using a tumor-specific cDNA microarray predicts the prognosis of intermediate risk neuroblastomas. Cancer Cell 7: 337–350.