Advertisement

Acta Neuropathologica

, Volume 128, Issue 5, pp 723–732 | Cite as

The tumor suppressor prostate apoptosis response-4 (Par-4) is regulated by mutant IDH1 and kills glioma stem cells

  • Yinxing Liu
  • Misty R. Gilbert
  • Natasha Kyprianou
  • Vivek M. Rangnekar
  • Craig Horbinski
Original Paper

Abstract

Prostate apoptosis response-4 (Par-4) is an endogenous tumor suppressor that selectively induces apoptosis in a variety of cancers. Although it has been the subject of intensive research in other cancers, less is known about its significance in gliomas, including whether it is regulated by key driver mutations, has therapeutic potential against glioma stem cells (GSCs), and/or is a prognostic marker. We found that patient-derived gliomas with mutant isocitrate dehydrogenase 1 have markedly lower Par-4 expression (P < 0.0001), which was validated by The Cancer Genome Atlas dataset (P = 2.0 E-13). The metabolic product of mutant IDH1, D-2-hydroxyglutarate (2-HG), can suppress Par-4 transcription in vitro via inhibition of promoter activity as well as enhanced mRNA degradation, but interestingly not by direct DNA promoter hypermethylation. The Selective for Apoptosis induction in Cancer cells (SAC) domain within Par-4 is highly active against glioma cells, including orthotopic xenografts of patient-derived primary GSCs (P < 0.0001). Among high-grade gliomas that are IDH1 wild type, those that express more Par-4 have significantly longer median survival (18.4 vs. 8.0 months, P = 0.002), a finding confirmed in two external GBM cohorts. Together, these data suggest that Par-4 is a significant component of the mutant IDH1 phenotype, that the activity of 2-HG is complex and can extend beyond direct DNA hypermethylation, and that Par-4 is a promising therapeutic strategy against GSCs. Furthermore, not every effect of mutant IDH1 necessarily contributes to the overall favorable prognosis seen in such tumors; inhibition of Par-4 may be one such effect.

Keywords

IDH1 Glioma Par-4 Tumor prognosis Glioma stem cells 

Notes

Acknowledgments

CH was supported by K08 CA155764 (National Cancer Institute), 2P20 RR020171 COBRE pilot grant (National Institute of General Medical Sciences), The Peter and Carmen Lucia Buck Training Program in Translational Clinical Oncology, and the University of Kentucky College of Medicine Physician Scientist Program. This work was also supported by NIH R01 CA060872 (to VMR). This research was supported by the Biospecimen and Tissue Procurement Shared Resource Facility of the University of Kentucky Markey Cancer Center (P30CA177558). Special thanks to Dana Napier for her excellent histologic expertise, and to Ravshan Burikhanov for his technical assistance. We thank Dr. Hai Yan of Duke University Medical Center for supplying us with the pEGFP-N1-IDH1R132H plasmid, and Drs. Jeremy Rich and Monica Venere of the Cleveland Clinic for patient-derived GBM stem cells. We also thank Dr. Chunming Liu of the University of Kentucky for the renilla luciferase plasmid.

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

401_2014_1334_MOESM1_ESM.pdf (425 kb)
Supplementary material 1 (PDF 424 kb)

References

  1. 1.
    Alvarez JV, Pan TC, Ruth J et al (2013) Par-4 downregulation promotes breast cancer recurrence by preventing multinucleation following targeted therapy. Cancer Cell 24(1):30–44PubMedCrossRefGoogle Scholar
  2. 2.
    Auffinger B, Tobias AL, Han Y et al (2014) Conversion of differentiated cancer cells into cancer stem-like cells in a glioblastoma model after primary chemotherapy. Cell Death Differ 21(7):1119–1131PubMedCrossRefGoogle Scholar
  3. 3.
    Azmi AS, Philip PA, Zafar SF, Sarkar FH, Mohammad RM (2010) PAR-4 as a possible new target for pancreatic cancer therapy. Expert Opin Ther Targets 14(6):611–620PubMedPubMedCentralCrossRefGoogle Scholar
  4. 4.
    Bao S, Wu Q, McLendon RE et al (2006) Glioma stem cells promote radioresistance by preferential activation of the DNA damage response. Nature 444(7120):756–760PubMedCrossRefGoogle Scholar
  5. 5.
    Barradas M, Monjas A, Diaz-Meco MT, Serrano M, Moscat J (1999) The downregulation of the pro-apoptotic protein Par-4 is critical for Ras-induced survival and tumor progression. EMBO J 18(22):6362–6369PubMedPubMedCentralCrossRefGoogle Scholar
  6. 6.
    Belasco J, Browerman G (1993) Control of messenger RNA stability. Academic Press, San DiegoGoogle Scholar
  7. 7.
    Berra E, Municio MM, Sanz L, Frutos S, Diaz-Meco MT, Moscat J (1997) Positioning atypical protein kinase C isoforms in the UV-induced apoptotic signaling cascade. Mol Cell Biol 17(8):4346–4354PubMedPubMedCentralGoogle Scholar
  8. 8.
    Boehrer S, Chow KU, Beske F et al (2002) In lymphatic cells par-4 sensitizes to apoptosis by down-regulating bcl-2 and promoting disruption of mitochondrial membrane potential and caspase activation. Cancer Res 62(6):1768–1775PubMedGoogle Scholar
  9. 9.
    Borodovsky A, Salmasi V, Turcan S et al (2013) 5-azacytidine reduces methylation, promotes differentiation and induces tumor regression in a patient-derived IDH1 mutant glioma xenograft. Oncotarget 4(10):1737–1747PubMedPubMedCentralGoogle Scholar
  10. 10.
    Burikhanov R, Zhao Y, Goswami A, Qiu S, Schwarze SR, Rangnekar VM (2009) The tumor suppressor Par-4 activates an extrinsic pathway for apoptosis. Cell 138(2):377–388PubMedPubMedCentralCrossRefGoogle Scholar
  11. 11.
    Capper D, Gaiser T, Hartmann C et al (2009) Stem-cell-like glioma cells are resistant to TRAIL/Apo2L and exhibit down-regulation of caspase-8 by promoter methylation. Acta Neuropathol 117(4):445–456PubMedCrossRefGoogle Scholar
  12. 12.
    Chakraborty M, Qiu SG, Vasudevan KM, Rangnekar VM (2001) Par-4 drives trafficking and activation of Fas and Fasl to induce prostate cancer cell apoptosis and tumor regression. Cancer Res 61(19):7255–7263PubMedGoogle Scholar
  13. 13.
    Chang S, Kim JH, Shin J (2002) p62 forms a ternary complex with PKCzeta and PAR-4 and antagonizes PAR-4-induced PKCzeta inhibition. FEBS Lett 510(1–2):57–61PubMedCrossRefGoogle Scholar
  14. 14.
    Chesnelong C, Chaumeil MM, Blough MD et al (2013) Lactate dehydrogenase A silencing in IDH mutant gliomas. Neuro Oncol 16(5):686–695PubMedCrossRefGoogle Scholar
  15. 15.
    Chou AP, Chowdhury R, Li S et al (2012) Identification of retinol binding protein 1 promoter hypermethylation in isocitrate dehydrogenase 1 and 2 mutant gliomas. J Natl Cancer Inst 104(19):1458–1469PubMedPubMedCentralCrossRefGoogle Scholar
  16. 16.
    Cohen AL, Holmen SL, Colman H (2013) IDH1 and IDH2 mutations in gliomas. Curr Neurol Neurosci Rep 13(5):345PubMedPubMedCentralCrossRefGoogle Scholar
  17. 17.
    Coutinho-Camillo CM, Lourenco SV, Nonogaki S et al (2013) Expression of PAR-4 and PHLDA1 is prognostic for overall and disease-free survival in oral squamous cell carcinomas. Virchows Arch 463(1):31–39PubMedCrossRefGoogle Scholar
  18. 18.
    Dang L, White DW, Gross S et al (2009) Cancer-associated IDH1 mutations produce 2-hydroxyglutarate. Nature 462(7274):739–744PubMedPubMedCentralCrossRefGoogle Scholar
  19. 19.
    Dey M, Ulasov IV, Tyler MA, Sonabend AM, Lesniak MS (2011) Cancer stem cells: the final frontier for glioma virotherapy. Stem Cell Rev 7(1):119–129PubMedPubMedCentralCrossRefGoogle Scholar
  20. 20.
    Diaz-Meco MT, Municio MM, Frutos S et al (1996) The product of par-4, a gene induced during apoptosis, interacts selectively with the atypical isoforms of protein kinase C. Cell 86(5):777–786PubMedCrossRefGoogle Scholar
  21. 21.
    El-Guendy N, Zhao Y, Gurumurthy S, Burikhanov R, Rangnekar VM (2003) Identification of a unique core domain of par-4 sufficient for selective apoptosis induction in cancer cells. Mol Cell Biol 23(16):5516–5525PubMedPubMedCentralCrossRefGoogle Scholar
  22. 22.
    Fernandez-Marcos PJ, Abu-Baker S, Joshi J et al (2009) Simultaneous inactivation of Par-4 and PTEN in vivo leads to synergistic NF-kappaB activation and invasive prostate carcinoma. Proc Natl Acad Sci USA 106(31):12962–12967PubMedPubMedCentralCrossRefGoogle Scholar
  23. 23.
    Figueroa ME, Abdel-Wahab O, Lu C et al (2010) Leukemic IDH1 and IDH2 mutations result in a hypermethylation phenotype, disrupt TET2 function, and impair hematopoietic differentiation. Cancer Cell 18(6):553–567PubMedPubMedCentralCrossRefGoogle Scholar
  24. 24.
    Garcia-Cao I, Duran A, Collado M et al (2005) Tumour-suppression activity of the proapoptotic regulator Par4. EMBO Rep 6(6):577–583PubMedPubMedCentralCrossRefGoogle Scholar
  25. 25.
    Gilbert MR, Liu Y, Neltner J et al (2014) Autophagy and oxidative stress in gliomas with IDH1 mutations. Acta Neuropathol 127(2):221–233PubMedCrossRefGoogle Scholar
  26. 26.
    Goswami A, Burikhanov R, de Thonel A et al (2005) Binding and phosphorylation of par-4 by akt is essential for cancer cell survival. Mol Cell 20(1):33–44PubMedCrossRefGoogle Scholar
  27. 27.
    Goswami A, Qiu S, Dexheimer TS et al (2008) Par-4 binds to topoisomerase 1 and attenuates its DNA relaxation activity. Cancer Res 68(15):6190–6198PubMedPubMedCentralCrossRefGoogle Scholar
  28. 28.
    Horbinski C (2013) What do we know about IDH1/2 mutations so far, and how do we use it? Acta Neuropathol 125(5):621–636PubMedPubMedCentralCrossRefGoogle Scholar
  29. 29.
    Jagtap JC, Dawood P, Shah RD et al (2014) Expression and regulation of prostate apoptosis response-4 (par-4) in human glioma stem cells in drug-induced apoptosis. PLoS One 9(2):e88505PubMedPubMedCentralCrossRefGoogle Scholar
  30. 30.
    Joshi J, Fernandez-Marcos PJ, Galvez A et al (2008) Par-4 inhibits Akt and suppresses Ras-induced lung tumorigenesis. EMBO J 27(16):2181–2193PubMedPubMedCentralCrossRefGoogle Scholar
  31. 31.
    Lafuente MJ, Martin P, Garcia-Cao I, Diaz-Meco MT, Serrano M, Moscat J (2003) Regulation of mature T lymphocyte proliferation and differentiation by Par-4. EMBO J 22(18):4689–4698PubMedPubMedCentralCrossRefGoogle Scholar
  32. 32.
    Lee TJ, Jang JH, Noh HJ, Park EJ, Choi KS, Kwon TK (2010) Overexpression of Par-4 sensitizes TRAIL-induced apoptosis via inactivation of NF-kappaB and Akt signaling pathways in renal cancer cells. J Cell Biochem 109(5):885–895PubMedGoogle Scholar
  33. 33.
    Lee TJ, Lee JT, Kim SH et al (2008) Overexpression of Par-4 enhances thapsigargin-induced apoptosis via down-regulation of XIAP and inactivation of Akt in human renal cancer cells. J Cell Biochem 103(2):358–368PubMedCrossRefGoogle Scholar
  34. 34.
    Li S, Chou AP, Chen W et al (2013) Overexpression of isocitrate dehydrogenase mutant proteins renders glioma cells more sensitive to radiation. Neuro Oncol 15(1):57–68PubMedPubMedCentralCrossRefGoogle Scholar
  35. 35.
    Liu J, Albrecht AM, Ni X, Yang J, Li M (2013) Glioblastoma tumor initiating cells: therapeutic strategies targeting apoptosis and microRNA pathways. Curr Mol Med 13(3):352–357PubMedGoogle Scholar
  36. 36.
    Lu C, Li JY, Ge Z, Zhang L, Zhou GP (2013) Par-4/THAP1 complex and Notch3 competitively regulated pre-mRNA splicing of CCAR1 and affected inversely the survival of T-cell acute lymphoblastic leukemia cells. Oncogene 32(50):5602–5613PubMedPubMedCentralCrossRefGoogle Scholar
  37. 37.
    Lu C, Venneti S, Akalin A et al (2013) Induction of sarcomas by mutant IDH2. Genes Dev 27(18):1986–1998PubMedPubMedCentralCrossRefGoogle Scholar
  38. 38.
    Lu C, Ward PS, Kapoor GS et al (2012) IDH mutation impairs histone demethylation and results in a block to cell differentiation. Nature 483(7390):474–478PubMedPubMedCentralCrossRefGoogle Scholar
  39. 39.
    Lucas T, Pratscher B, Krishnan S et al (2001) Differential expression levels of Par-4 in melanoma. Melanoma Res 11(4):379–383PubMedCrossRefGoogle Scholar
  40. 40.
    Mayers JR, Vander Heiden MG (2013) BCAT1 defines gliomas by IDH status. Nat Med 19(7):816–817PubMedCrossRefGoogle Scholar
  41. 41.
    Nagai MA, Gerhard R, Salaorni S et al (2010) Down-regulation of the candidate tumor suppressor gene PAR-4 is associated with poor prognosis in breast cancer. Int J Oncol 37(1):41–49PubMedCrossRefGoogle Scholar
  42. 42.
    Ohba S, Mukherjee J, See WL, Pieper RO (2014) Mutant IDH1-driven cellular transformation increases RAD51-mediated homologous recombination and temozolomide resistance. Cancer Res (Epub ahead of print)Google Scholar
  43. 43.
    Pereira MC, de Bessa-Garcia SA, Burikhanov R et al (2013) Prostate apoptosis response-4 is involved in the apoptosis response to docetaxel in MCF-7 breast cancer cells. Int J Oncol 43(2):531–538PubMedPubMedCentralGoogle Scholar
  44. 44.
    Qiu G, Ahmed M, Sells SF, Mohiuddin M, Weinstein MH, Rangnekar VM (1999) Mutually exclusive expression patterns of Bcl-2 and Par-4 in human prostate tumors consistent with down-regulation of Bcl-2 by Par-4. Oncogene 18(3):623–631PubMedCrossRefGoogle Scholar
  45. 45.
    Qiu SG, Krishnan S, el-Guendy N, Rangnekar VM (1999) Negative regulation of Par-4 by oncogenic Ras is essential for cellular transformation. Oncogene 18(50):7115–7123PubMedCrossRefGoogle Scholar
  46. 46.
    Reitman ZJ, Jin G, Karoly ED et al (2011) Profiling the effects of isocitrate dehydrogenase 1 and 2 mutations on the cellular metabolome. Proc Natl Acad Sci USA 108(8):3270–3275PubMedPubMedCentralCrossRefGoogle Scholar
  47. 47.
    Rohle D, Popovici-Muller J, Palaskas N et al (2013) An inhibitor of mutant IDH1 delays growth and promotes differentiation of glioma cells. Science 340(6132):626–630PubMedPubMedCentralCrossRefGoogle Scholar
  48. 48.
    Saegusa M, Hashimura M, Kuwata T, Okayasu I (2010) Transcriptional regulation of pro-apoptotic Par-4 by NF-kappaB/p65 and its function in controlling cell kinetics during early events in endometrial tumourigenesis. J Pathol 221(1):26–36PubMedCrossRefGoogle Scholar
  49. 49.
    Sells SF, Han SS, Muthukkumar S et al (1997) Expression and function of the leucine zipper protein Par-4 in apoptosis. Mol Cell Biol 17(7):3823–3832PubMedPubMedCentralGoogle Scholar
  50. 50.
    Sharma AK, Kline CL, Berg A, Amin S, Irby RB (2011) The Akt inhibitor ISC-4 activates prostate apoptosis response protein-4 and reduces colon tumor growth in a nude mouse model. Clin Cancer Res 17(13):4474–4483PubMedPubMedCentralCrossRefGoogle Scholar
  51. 51.
    Shrestha-Bhattarai T, Rangnekar VM (2010) Cancer-selective apoptotic effects of extracellular and intracellular Par-4. Oncogene 29(27):3873–3880PubMedCrossRefGoogle Scholar
  52. 52.
    Srinivasan S, Ranga RS, Burikhanov R, Han SS, Chendil D (2007) Par-4-dependent apoptosis by the dietary compound withaferin A in prostate cancer cells. Cancer Res 67(1):246–253PubMedCrossRefGoogle Scholar
  53. 53.
    Vetterkind S, Boosen M, Scheidtmann KH et al (2005) Ectopic expression of Par-4 leads to induction of apoptosis in CNS tumor cell lines. Int J Oncol 26(1):159–167PubMedGoogle Scholar
  54. 54.
    Wei J, Barr J, Kong LY et al (2010) Glioblastoma cancer-initiating cells inhibit T-cell proliferation and effector responses by the signal transducers and activators of transcription 3 pathway. Mol Cancer Ther 9(1):67–78PubMedPubMedCentralCrossRefGoogle Scholar
  55. 55.
    Wei J, Barr J, Kong LY et al (2010) Glioma-associated cancer-initiating cells induce immunosuppression. Clin Cancer Res 16(2):461–473PubMedPubMedCentralCrossRefGoogle Scholar
  56. 56.
    Wu A, Wei J, Kong LY et al (2010) Glioma cancer stem cells induce immunosuppressive macrophages/microglia. Neuro Oncol 12(11):1113–1125PubMedPubMedCentralCrossRefGoogle Scholar
  57. 57.
    Zhao Y, Burikhanov R, Brandon J et al (2011) Systemic Par-4 inhibits non-autochthonous tumor growth. Cancer Biol Ther 12(2):152–157PubMedPubMedCentralCrossRefGoogle Scholar
  58. 58.
    Zhao Y, Burikhanov R, Qiu S et al (2007) Cancer resistance in transgenic mice expressing the SAC module of Par-4. Cancer Res 67(19):9276–9285PubMedCrossRefGoogle Scholar
  59. 59.
    Zhuang D, Liu Y, Mao Y et al (2012) TMZ-induced PrPc/par-4 interaction promotes the survival of human glioma cells. Int J Cancer 130(2):309–318PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2014

Authors and Affiliations

  • Yinxing Liu
    • 1
  • Misty R. Gilbert
    • 1
  • Natasha Kyprianou
    • 1
    • 2
    • 5
  • Vivek M. Rangnekar
    • 3
    • 4
    • 5
  • Craig Horbinski
    • 1
    • 2
    • 5
  1. 1.Department of Pathology and Laboratory MedicineUniversity of Kentucky, 307 Combs BuildingLexingtonUSA
  2. 2.Department of Molecular and Cellular BiochemistryUniversity of KentuckyLexingtonUSA
  3. 3.Department of Radiation MedicineUniversity of KentuckyLexingtonUSA
  4. 4.Department of Microbiology, Immunology and Molecular GeneticsUniversity of KentuckyLexingtonUSA
  5. 5.Markey Cancer CenterUniversity of KentuckyLexingtonUSA

Personalised recommendations