Skip to main content
SpringerLink
Log in

Germ-line PHD1 and PHD2 mutations detected in patients with pheochromocytoma/paraganglioma-polycythemia

  • Original Article
  • Published:
Journal of Molecular Medicine Aims and scope Submit manuscript

Abstract

We have investigated genetic/pathogenetic factors associated with a new clinical entity in patients presenting with pheochromocytoma/paraganglioma (PHEO/PGL) and polycythemia. Two patients without hypoxia-inducible factor 2α (HIF2A) mutations, who presented with similar clinical manifestations, were analyzed for other gene mutations, including prolyl hydroxylase (PHD) mutations. We have found for the first time a germ-line mutation in PHD1 in one patient and a novel germ-line PHD2 mutation in a second patient. Both mutants exhibited reduced protein stability with substantial quantitative protein loss and thus compromised catalytic activities. Due to the unique association of patients’ polycythemia with borderline or mildly elevated erythropoietin (EPO) levels, we also performed an in vitro sensitivity assay of erythroid progenitors to EPO and for EPO receptor (EPOR) expression. The results show inappropriate hypersensitivity of erythroid progenitors to EPO in these patients, indicating increased EPOR expression/activity. In addition, the present study indicates that HIF dysregulation due to PHD mutations plays an important role in the pathogenesis of these tumors and associated polycythemia. The PHD1 mutation appears to be a new member contributing to the genetic landscape of this novel clinical entity. Our results support the existence of a specific PHD1- and PHD2-associated PHEO/PGL-polycythemia disorder.

Key message

• A novel germ-line PHD1 mutation causing pheochromocytoma/paraganglioma and polycythemia.

• Increased EPOR activity and inappropriate hypersensitivity of erythroid progenitors to EPO.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

References

  1. Semenza GL (2012) Hypoxia-inducible factors in physiology and medicine. Cell 148:399–408

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  2. Jaakkola P, Mole DR, Tian YM, Wilson MI, Gielbert J, Gaskell SJ, von Kriegsheim A, Hebestreit HF, Mukherji M, Schofield CJ et al (2001) Targeting of HIF-alpha to the von Hippel-Lindau ubiquitylation complex by O2-regulated prolyl hydroxylation. Science 292:468–472

    Article  CAS  PubMed  Google Scholar 

  3. Ivan M, Haberberger T, Gervasi DC, Michelson KS, Gunzler V, Kondo K, Yang H, Sorokina I, Conaway RC, Conaway JW et al (2002) Biochemical purification and pharmacological inhibition of a mammalian prolyl hydroxylase acting on hypoxia-inducible factor. Proc Natl Acad Sci U S A 99:13459–13464

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  4. Kaelin WG Jr (2008) The von Hippel-Lindau tumour suppressor protein: O2 sensing and cancer. Nat Rev Cancer 8:865–873

    Article  CAS  PubMed  Google Scholar 

  5. Scortegagna M, Ding K, Zhang Q, Oktay Y, Bennett MJ, Bennett M, Shelton JM, Richardson JA, Moe O, Garcia JA (2005) HIF-2alpha regulates murine hematopoietic development in an erythropoietin-dependent manner. Blood 105:3133–3140

    Article  CAS  PubMed  Google Scholar 

  6. Prchal JT (2005) Polycythemia vera and other primary polycythemias. Curr Opin Hematol 12:112–116

    Article  PubMed  Google Scholar 

  7. McMullin MF (2010) HIF pathway mutations and erythrocytosis. Expert Rev Hematol 3:93–101

    Article  CAS  PubMed  Google Scholar 

  8. Ang SO, Chen H, Hirota K, Gordeuk VR, Jelinek J, Guan Y, Liu E, Sergueeva AI, Miasnikova GY, Mole D et al (2002) Disruption of oxygen homeostasis underlies congenital Chuvash polycythemia. Nat Genet 32:614–621

    Article  CAS  PubMed  Google Scholar 

  9. Percy MJ, Beer PA, Campbell G, Dekker AW, Green AR, Oscier D, Rainey MG, van Wijk R, Wood M, Lappin TR et al (2008) Novel exon 12 mutations in the HIF2A gene associated with erythrocytosis. Blood 111:5400–5402

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  10. Prchal JT, Gordeuk VR (2008) The HIF2A gene in familial erythrocytosis. N Engl J Med 358:1966, author reply 1966–1967

    CAS  PubMed  Google Scholar 

  11. de la Chapelle A, Traskelin AL, Juvonen E (1993) Truncated erythropoietin receptor causes dominantly inherited benign human erythrocytosis. Proc Natl Acad Sci U S A 90:4495–4499

    Article  PubMed Central  PubMed  Google Scholar 

  12. Sokol L, Luhovy M, Guan Y, Prchal JF, Semenza GL, Prchal JT (1995) Primary familial polycythemia: a frameshift mutation in the erythropoietin receptor gene and increased sensitivity of erythroid progenitors to erythropoietin. Blood 86:15–22

    CAS  PubMed  Google Scholar 

  13. Albiero E, Ruggeri M, Fortuna S, Finotto S, Bernardi M, Madeo D, Rodeghiero F (2012) Isolated erythrocytosis: study of 67 patients and identification of three novel germ-line mutations in the prolyl hydroxylase domain protein 2 (PHD2) gene. Haematologica 97:123–127

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  14. Lenders JW, Eisenhofer G, Mannelli M, Pacak K (2005) Phaeochromocytoma. Lancet 366:665–675

    Article  PubMed  Google Scholar 

  15. Ladroue C, Carcenac R, Leporrier M, Gad S, Le Hello C, Galateau-Salle F, Feunteun J, Pouyssegur J, Richard S, Gardie B (2008) PHD2 mutation and congenital erythrocytosis with paraganglioma. N Engl J Med 359:2685–2692

    Article  CAS  PubMed  Google Scholar 

  16. Zhuang Z, Yang C, Lorenzo F, Merino M, Fojo T, Kebebew E, Popovic V, Stratakis CA, Prchal JT, Pacak K (2012) Somatic HIF2A gain-of-function mutations in paraganglioma with polycythemia. N Engl J Med 367:922–930

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  17. Capodimonti S, Teofili L, Martini M, Cenci T, Iachininoto MG, Nuzzolo ER, Bianchi M, Murdolo M, Leone G, Larocca LM (2012) Von hippel-lindau disease and erythrocytosis. J Clin Oncol 30:e137–e139

    Article  PubMed  Google Scholar 

  18. Lorenzo FR, Yang C, Ng Tang Fui M, Vankayalapati H, Zhuang Z, Huynh T, Grossmann M, Pacak K, Prchal JT (2013) A novel EPAS1/HIF2A germline mutation in a congenital polycythemia with paraganglioma. J Mol Med (Berl) 91:507–512

    Article  CAS  Google Scholar 

  19. Yang C, Huntoon K, Ksendzovsky A, Zhuang Z, Lonser RR (2013) Proteostasis modulators prolong missense VHL protein activity and halt tumor progression. Cell Rep 3:52–59

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  20. Shankavaram U, Fliedner SM, Elkahloun AG, Barb JJ, Munson PJ, Huynh TT, Matro JC, Turkova H, Linehan WM, Timmers HJ et al (2013) Genotype and tumor locus determine expression profile of pseudohypoxic pheochromocytomas and paragangliomas. Neoplasia 15:435–447

    CAS  PubMed Central  PubMed  Google Scholar 

  21. Schuetz CS, Bonin M, Clare SE, Nieselt K, Sotlar K, Walter M, Fehm T, Solomayer E, Riess O, Wallwiener D et al (2006) Progression-specific genes identified by expression profiling of matched ductal carcinomas in situ and invasive breast tumors, combining laser capture microdissection and oligonucleotide microarray analysis. Cancer Res 66:5278–5286

    Article  CAS  PubMed  Google Scholar 

  22. Lanikova L, Lorenzo F, Yang C, Vankayalapati H, Drachtman R, Divoky V, Prchal JT (2013) Novel homozygous VHL mutation in exon 2 is associated with congenital polycythemia but not with cancer. Blood 121:3918–3924

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  23. Jochmanova I, Yang C, Zhuang Z, Pacak K (2013) Hypoxia-inducible factor signaling in pheochromocytoma: turning the rudder in the right direction. J Natl Cancer Inst 105:1270–1283

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  24. Jokilehto T, Jaakkola PM (2010) The role of HIF prolyl hydroxylases in tumour growth. J Cell Mol Med 14:758–770

    Article  CAS  PubMed  Google Scholar 

  25. Selak MA, Armour SM, MacKenzie ED, Boulahbel H, Watson DG, Mansfield KD, Pan Y, Simon MC, Thompson CB, Gottlieb E (2005) Succinate links TCA cycle dysfunction to oncogenesis by inhibiting HIF-alpha prolyl hydroxylase. Cancer Cell 7:77–85

    Article  CAS  PubMed  Google Scholar 

  26. Takeda K, Fong GH (2007) Prolyl hydroxylase domain 2 protein suppresses hypoxia-induced endothelial cell proliferation. Hypertension 49:178–184

    Article  CAS  PubMed  Google Scholar 

  27. Tian YM, Mole DR, Ratcliffe PJ, Gleadle JM (2006) Characterization of different isoforms of the HIF prolyl hydroxylase PHD1 generated by alternative initiation. Biochem J 397:179–186

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  28. Arsenault PR, Pei F, Lee R, Kerestes H, Percy MJ, Keith B, Simon MC, Lappin TR, Khurana TS, Lee FS (2013) A knock-in mouse model of human PHD2 gene-associated erythrocytosis establishes a haploinsufficiency mechanism. J Biol Chem 288:33571–33584

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  29. Juul SE, Yachnis AT, Christensen RD (1998) Tissue distribution of erythropoietin and erythropoietin receptor in the developing human fetus. Early Hum Dev 52:235–249

    Article  CAS  PubMed  Google Scholar 

  30. Miyake M, Goodison S, Lawton A, Zhang G, Gomes-Giacoia E, Rosser CJ (2013) Erythropoietin is a JAK2 and ERK1/2 effector that can promote renal tumor cell proliferation under hypoxic conditions. J Hematol Oncol 6:65

    Article  PubMed Central  PubMed  Google Scholar 

  31. Shi Z, Hodges VM, Dunlop EA, Percy MJ, Maxwell AP, El-Tanani M, Lappin TR (2010) Erythropoietin-induced activation of the JAK2/STAT5, PI3K/Akt, and Ras/ERK pathways promotes malignant cell behavior in a modified breast cancer cell line. Mol Cancer Res 8:615–626

    Article  CAS  PubMed  Google Scholar 

  32. Um M, Gross AW, Lodish HF (2007) A “classical” homodimeric erythropoietin receptor is essential for the antiapoptotic effects of erythropoietin on differentiated neuroblastoma SH-SY5Y and pheochromocytoma PC-12 cells. Cell Signal 19:634–645

    Article  CAS  PubMed  Google Scholar 

  33. Yoon D, Agarwal N, Prchal JT (2008) Does erythropoietin promote tumor growth? Clin Cancer Res 14:1920, author reply 1920–1921

    Article  CAS  PubMed  Google Scholar 

  34. Hu CJ, Wang LY, Chodosh LA, Keith B, Simon MC (2003) Differential roles of hypoxia-inducible factor 1alpha (HIF-1alpha) and HIF-2alpha in hypoxic gene regulation. Mol Cell Biol 23:9361–9374

    Article  CAS  PubMed Central  PubMed  Google Scholar 

Download references

Acknowledgments

We would like to acknowledge the clinical and technical assistance of Victoria Martucci, Karen Adams, Pauline Dmitriev, and Joan Nambuba in the production of this manuscript. We also thank both patients and their families for their participation and assistance. This research was supported, in part, by the Intramural Research Program of the NIH, Eunice Kennedy Shriver NICHD, NINDS, NCI, and NHGRI.

Conflict of interest

The authors declare no conflict of interests related to this study.

Author information

Authors and Affiliations

  1. Surgical Neurology Branch, National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, MD, USA

    Chunzhang Yang, Zhengping Zhuang, Michael G. Sun & Roland Zhu

  2. 1st Department of Medicine, University Medical Center Schleswig-Holstein, Lübeck, Germany

    Stephanie M. J. Fliedner

  3. Radiation Oncology Branch, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA

    Uma Shankavaram

  4. Program in Reproductive and Adult Endocrinology, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Building 10, CRC, Room 1E-3140, 10 Center Drive, MSC-1109, Bethesda, MD, 20892-1109, USA

    Petra Bullova & Karel Pacak

  5. Department of Molecular Medicine, Institute of Virology, Slovak Academy of Sciences, Bratislava, Slovakia

    Petra Bullova

  6. Center for Research on Genomics and Global Health, National Human Genome Research Institute, National Institutes of Health, Bethesda, MD, USA

    Abdel G. Elkahloun

  7. Columbus Hematology and Oncology Associates, Columbus, OH, USA

    Peter J. Kourlas

  8. Laboratory of Pathology, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA

    Maria Merino

  9. Endocrine Oncology Branch, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA

    Electron Kebebew

Authors
  1. Chunzhang Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Zhengping Zhuang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Stephanie M. J. Fliedner
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Uma Shankavaram
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Michael G. Sun
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Petra Bullova
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Roland Zhu
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Abdel G. Elkahloun
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Peter J. Kourlas
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Maria Merino
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Electron Kebebew
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Karel Pacak
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Karel Pacak.

Additional information

Chunzhang Yang and Zhengping Zhuang contributed equally to this article.

Electronic supplementary material

Below is the link to the electronic supplementary material.

ESM 1

(PDF 6711 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Yang, C., Zhuang, Z., Fliedner, S.M.J. et al. Germ-line PHD1 and PHD2 mutations detected in patients with pheochromocytoma/paraganglioma-polycythemia. J Mol Med 93, 93–104 (2015). https://doi.org/10.1007/s00109-014-1205-7

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00109-014-1205-7

Keywords

Search

Navigation