, Volume 19, Issue 4, pp 407–414 | Cite as

A high-throughput analysis of the IDH1(R132H) protein expression in pituitary adenomas

  • Olivera Casar-BorotaEmail author
  • Kristin Astrid Berland Øystese
  • Magnus Sundström
  • Linea Melchior
  • Vera Popovic



Inactivating mutations of isocitrate dehydrogenase (IDH) 1 and 2, mitochondrial enzymes participating in the Krebs tricarboxylic acid cycle play a role in the tumorigenesis of gliomas and also less frequently in acute myeloid leukemia and other malignancies. Inhibitors of mutant IDH1 and IDH2 may potentially be effective in the treatment of the IDH mutation driven tumors. Mutations in the succinate dehydrogenase, the other enzyme complex participating in the Krebs cycle and electron transfer of oxidative phosphorylation occur in the paragangliomas, gastrointestinal stromal tumors, and occasionally in the pituitary adenomas. We aimed to determine whether the IDH1(R132H) mutation, the most frequent IDH mutation in human malignancies, occurs in pituitary adenomas.


We performed immunohistochemical analysis by using a monoclonal anti-IDH1(R132H) antibody on the tissue microarrays containing specimens from the pituitary adenomas of different hormonal types from 246 patients. In positive samples, the status of the IDH1 gene was further examined by molecular genetic analyses.


In all but one patient, there was no expression of mutated IDH1(R132H) protein in the tumor cells by immunohistochemistry. Only one patient with a recurring clinically non-functioning gonadotroph adenoma demonstrated IDH1(R132H)-immunostaining in both the primary tumor and the recurrence. However, no mutation in the IDH1 gene was detected using different molecular genetic analyses.


IDH1(R132H) mutation occurs only exceptionally in pituitary adenomas and does not play a role in their pathogenesis. Patients with pituitary adenomas do not seem to be candidates for treatment with the inhibitors of mutant IDH1.


Pituitary adenoma Isocitrate dehydrogenase IDH1(R132H) Immunohistochemistry Tissue microarrays 



We thank Ellen Hellesylt for her assistance with the construction of the tissue microarrays and immunohistochemical analyses, Dijana Cerjan for her assistance with the immunohistochemistry, and Simona Cionca for her assistance with DNA extraction and MLPA analysis. Prof Dr. Vera Popovic was supported by the Ministry of Education and Science of Republic of Serbia, Grant No 175033.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.


  1. 1.
    Desideri E, Vegliante R, Ciriolo MR (2015) Mitochondrial dysfunctions in cancer: genetic defects and oncogenic signaling impinging on TCA cycle activity. Cancer Lett 356:217–223CrossRefPubMedGoogle Scholar
  2. 2.
    Parker SJ, Metallo CM (2015) Metabolic consequences of oncogenic IDH mutations. Pharmacol Ther 152:54–62CrossRefPubMedGoogle Scholar
  3. 3.
    Dang L, White DW, Gross S, Bennett BD, Bittinger MA, Driggers EM, Fantin VR, Jang HG, Jin S, Keenan MC, Marks KM, Prins RM, Ward PS, Yen KE, Liau LM, Rabinowitz JD, Cantley LC, Thompson CB, Heiden MGV, Su SM (2009) Cancer-associated IDH1 mutations produce 2-hydroxyglutarate. Nature 462:739–744CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Ward PS, Patel J, Wise DR, Abdel-Wahab O, Bennett BD, Coller HA, Cross JR, Fantin VR, Hedvat CV, Perl AE, Rabinowitz JD, Carroll M, Su SM, Sharp KA, Levine RL, Thompson CB (2010) The common feature of leukemia-associated IDH1 and IDH2 mutations is a neomorphic enzymatic activity that converts α-ketoglutarate to 2-hydroxyglutarate. Cancer Cell 17:225–234CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Lu C, Ward PS, Kapoor GS, Rohle D, Turcan S, Abdel-Wahab O, Edwards CR, Khanin R, Figueroa ME, Melnick A, Wellen KE, O’Rourke DM, Berger SL, Chan TA, Levine RL, Mellinghoff IK, Thompson CB (2012) IDH mutation impairs histone demethylation and results in a block to cell differentiation. Nature 483:474–478CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Oizel K, Gratas C, Nadaradjane A, Oliver L, Vallette FM, Pecqueur C (2015) D-2-Hydroxyglutarate does not mimic all the IDH mutation effects, in particular the reduced etoposide triggered apoptosis mediated by an alteration in mitochondrial NADH. Cell Death Dis. doi: 10.1038/cddis.2015.13 PubMedPubMedCentralGoogle Scholar
  7. 7.
    Zhao S, Lin Y, Xu W, Jiang W, Zha Z, Wang P, Yu W, Li Z, Gong L, Peng Y, Ding J, Lei Q, Guan K-L, Xiong Y (2009) Glioma-derived mutations in IDH1 dominantly inhibit IDH1 catalytic activity and induce HIF-1α. Science 324:261–265CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    Parsons DW, Jones S, Zhang X, Lin JC-H, Leary RJ, Angenendt P, Mankoo P, Carter H, Siu I-M, Gallia GL, Olivi A, McLendon R, Rasheed BA, Keir S, Nikolskaya T, Nikolsky Y, Busam DA, Tekleab H, Diaz LA Jr, Hartigan J, Smith DR, Strausberg RL, Marie SKN, Shinjo SMO, Yan H, Riggins GJ, Bigner DD, Karchin R, Papadopoulos N, Parmigiani G, Vogelstein B, Velculescu VE, Kinzler KW (2008) An integrated genomic analysis of human glioblastoma multiforme. Science 321:1807–1812CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Watanabe T, Nobusawa S, Kleihues P, Ohgaki H (2009) IDH1 mutations are early events in the development of astrocytomas and oligodendrogliomas. Am J Pathol 174:1149–1153CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Yan H, Parsons DW, Jin G, McLendon R, Rasheed BA, Yuan W, Kos I, Batinic-Haberle I, Jones S, Riggins GJ, Friedman H, Friedman A, Reardon D, Herndon J, Kinzler KW, Velculescu VE, Vogelstein B, Bigner DD (2009) IDH1 and IDH2 mutations in gliomas. N Engl J Med 360:765–773CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Mardis ER, Ding L, Dooling DJ, Larson DE, McLellan MD, Chen K, Koboldt DC, Fulton RS, Locke DP, Magrini VJ, Abbott RM, Vickery TL, Reed JS, Robinson JS, Wylie T, Smith SM, Carmichael L, Eldred JM, Harris CC, Walker J, Peck JB, Du F, Dukes AF, Sanderson GE, Brummett AM, Clark E, McMichael JF, Meyer RJ, Schindler JK, Pohl CS, Wallis JW, Shi X, Lin L, Schmidt H, Tang Y, Haipek C, Wiechert ME, Ivy JV, Kalicki J, Elliott G, Ries RE, Payton JE, Westervelt P, Tomasson MH, Watson MA, Baty J, Heath S, Shannon WD, Nagarajan R, Link DC, Walter MJ, Graubert TA, DiPersio JF, Wilson RK, Ley TJ (2009) Recurring mutations found by sequencing an acute lyeloid leukemia genome. N Engl J Med 361:1058–1066CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Amary MF, Bacsi K, Maggiani F, Damato S, Halai D, Berisha F, Pollock R, O’Donnell P, Grigoriadis A, Diss T, Eskandarpour M, Presneau N, Hogendoorn PCW, Futreal A, Tirabosco R, Flanagan AM (2011) IDH1 and IDH2 mutations are frequent events in central chondrosarcoma and central and periosteal chondromas but not in other mesenchymal tumours. J Pathol 224:334–343CrossRefPubMedGoogle Scholar
  13. 13.
    Borger DR, Tanabe KK, Fan KC, Lopez HU, Fantin VR, Straley KS, Schenkein DP, Hezel AF, Ancukiewicz M, Liebman HM, Kwak EL, Clark JW, Ryan DP, Deshpande V, Dias-Santagata D, Ellisen LW, Zhu AX, Iafrate AJ (2012) Frequent mutation of isocitrate dehydrogenase (IDH)1 and IDH2 in cholangiocarcinoma identified through broad-based tumor genotyping. Oncologist 17:72–79CrossRefPubMedGoogle Scholar
  14. 14.
    Preusser M, Capper D, Hartman C (2011) Euro-CNS Research Committee: IDH testing in diagnostic neuropathology: review and practical guideline article invited by the Euro-CNS research committee. Clin Neuropathol 30:217–230CrossRefPubMedGoogle Scholar
  15. 15.
    Rohle D, Popovici-Muller J, Palaskas N, Turcan S, Grommes C, Campos C, Tsoi J, Clark O, Oldrini B, Komisopoulou E, Kunii K, Pedraza A, Schalm S, Silverman L, Miller A, Wang F, Yang H, Chen Y, Kernytsky A, Rosenblum MK, Liu W, Biller SA, Su SM, Brennan CW, Chan TA, Graeber TG, Yen KE, Mellinghoff IK (2013) An inhibitor of mutant IDH1 delays growth and promotes differentiation of glioma cells. Science 340:626–630CrossRefPubMedPubMedCentralGoogle Scholar
  16. 16.
    Sasine JP, Schiller GJ (2015) Emerging strategies for high risk and relapsed/refractory acute myeloid leukemia: novel agents and approaches currently in clinical trials. Blood Rev 29:1–9CrossRefPubMedGoogle Scholar
  17. 17.
    Letouzé E, Martinelli C, Loriot C, Burnichon N, Abermil N, Ottolenghi C, Janin M, Menara M, Nguyen AT, Benit P, Buffet A, Marcaillou C, Bertherat J, Amar L, Rustin P, De Reyniés A, Gimenez-Roqueplo A-P, Favier J (2013) SDH mutations establish a hypermethylator phenotype in paraganglioma. Cancer Cell 23:739–752CrossRefPubMedGoogle Scholar
  18. 18.
    Gimenez-Roqueplo AP, Dahia PL, Robledo M (2012) An update on the genetics of paraganglioma, pheochromocytoma, and associated hereditary syndromes. Horm Metab Res 44:328–333CrossRefPubMedGoogle Scholar
  19. 19.
    Miettinen M, Lasota J (2014) Succinate dehydrogenase deficient gastrointestinal stromal tumors (GISTs)—a review. Int J Biochem Cell Biol 53:514–519CrossRefPubMedGoogle Scholar
  20. 20.
    Janeway KA, Kim SY, Lodish M, Nosé V, Rustin P, Gaal J, Dahia PL, Liegl B, Ball ER, Raygada M, Lai AH, Kelly L, Hornick JL, O’Sullivan M, de Krijger RR, Dinjens WNM, Demetri GD, Antonescu CR, Fletcher JA, Helman L, Stratakis CA (2011) Defects in succinate dehydrogenase in gastrointestinal stromal tumors lacking KIT and PDGFRA mutations. Proc Natl Acad Sci USA 108:314–318CrossRefPubMedGoogle Scholar
  21. 21.
    Ricketts C, Woodward ER, Killick P, Morris MR, Astuti D, Latif F, Maher ER (2008) Germline SDHB mutations and familial renal cell carcinoma. J Natl Cancer Inst 100:1260–1262CrossRefPubMedGoogle Scholar
  22. 22.
    Dénes J, Swords F, Rattenberry E, Stals K, Owens M, Cranston T, Xekouki P, Moran L, Kumar A, Wassif C, Fersht N, Baldeweg SE, Morris D, Lightman S, Agha A, Rees A, Grieve J, Powell M, Boguszewski CL, Dutta P, Thakker RV, Srirangalingam U, Thompson CJ, Druce M, Higham C, Davis J, Eeles R, Stevenson M, O’Sullivan B, Taniere P, Skordilis K, Gabrovska P, Barlier A, Webb SM, Aulinas A, Drake WM, Bevan JS, Preda C, Dalantaeva N, Ribeiro-Oliveira A Jr, Garcia IT, Yordanova G, Iotova V, Evanson J, Grossman AB, Trouillas J, Ellard S, Stratakis CA, Eamonn R, Maher EA, Roncaroli F, Korbonits M (2015) Heterogeneous genetic background of the association of pheochromocytoma/paraganglioma and pituitary adenoma: results from a large patient cohort. J Clin Endocrinol Metab 100:E531–E541CrossRefPubMedGoogle Scholar
  23. 23.
    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-1 prolyl hydroxylase. Cancer Cell 7:177–185CrossRefGoogle Scholar
  24. 24.
    King A, Selak MA, Gottlieb E (2006) Succinate dehydrogenase and fumarate hydratase: linking mitochondrial dysfunction and cancer. Oncogene 25:4675–4682CrossRefPubMedGoogle Scholar
  25. 25.
    Balss J, Meyer J, Mueller W, Korshunov A, Hartmann C, von Deimling A (2008) Analysis of the IDH1 codon 132 mutation in brain tumors. Acta Neuropathol 116:597–602CrossRefPubMedGoogle Scholar
  26. 26.
    Ikota H, Nobusawa S, Tanaka Y, Yokoo H, Nakazato Y (2011) High-throughput immunohistochemical profiling of primary brain tumors and non-neoplastic systemic organs with a specific antibody against the mutant isocitrate dehydrogenase 1 R132H protein. Brain Tumor Pathol 28:107–114CrossRefPubMedGoogle Scholar
  27. 27.
    Hao, S., Hong, C.S., Feng, J., Yang, C., Chittiboina, P., Zhang, J., Zhuang, Z.: Somatic IDH1 mutation in a pituitary adenoma of a patient with Maffucci syndrome. J Neurosurg. PMID: 26473790 (Epub) (2015)Google Scholar
  28. 28.
    Casar-Borota O, Fougner SL, Bollerslev J, Nesland JM (2012) KIT protein expression and mutational status of KIT gene in pituitary adenomas. Virchows Arch 460:171–181CrossRefPubMedGoogle Scholar
  29. 29.
    Capper D, Zentgraf H, Balss J, Hartmann C, von Deimling A (2009) Monoclonal antibody specific for IDH1 R132H mutation. Acta Neuropathol 118:599–601CrossRefPubMedGoogle Scholar
  30. 30.
    Capper D, Weißert S, Balss J, Habel A, Meyer J, Jäger D, Ackermann U, Tessmer C, Korshunov A, Zentgraf H, Hartmann C, von Deimling A (2010) Characterization of R132H mutation-specific IDH1 antibody binding in brain tumors. Brain Pathol 20:245–254CrossRefPubMedGoogle Scholar
  31. 31.
    Preusser M, Wöhrer A, Stary S, Höftberger R, Streubel B, Hainfellner JA (2011) Value and limitations of immunohistochemistry and gene sequencing for detection of the IDH1-R132H mutation in diffuse glioma biopsy specimens. J Neuropathol Exp Neurol 70:715–723CrossRefPubMedGoogle Scholar
  32. 32.
    Agarwal S, Sharma MC, Jha P, Pathak P, Suri V, Sarkar C, Chosdol K, Suri A, Kale SS, Mahapatra AK, Jha P (2013) Comparative study of IDH1 mutations in gliomas by immunohistochemistry and DNA sequencing. Neuro Oncol 15:718–726CrossRefPubMedPubMedCentralGoogle Scholar
  33. 33.
    Van den Bent MJ, Hartmann C, Preusser M, Ströbel T, Dubbink HJ, Kros JM, von Deimling A, Boisselier B, Sanson M, Halling KC, Diefes KL, Aldape K, Giannini C (2013) Interlaboratory comparison of IDH mutation detection. J Neurooncol 112:173–178CrossRefPubMedGoogle Scholar
  34. 34.
    Gaal J, Burnichon N, Korpershoek E, Roncelin I, Bertherat J, Plouin P-F, de Krijger RR, Gimenez-Roqueplo A-P, Dinjens WNM (2010) Isocitrate dehydrogenase mutations are rare in pheochromocytomas and paragangliomas. J Clin Endocrinol Metab 95:1274–1278CrossRefPubMedGoogle Scholar
  35. 35.
    Pai R, Manipadam MT, Singh P, Ebenazer A, Samuel P, Rajaratnam S (2014) Usefulness of succinate dehydrogenase B (SDHB) immunohistochemistry in guiding mutational screening among patients with pheochromocytoma-paraganglioma syndromes. APMIS. 122:1130–1135PubMedGoogle Scholar
  36. 36.
    Papathomas TG, Oudijk L, Persu A, Gill AJ, van Nederveen F, Tischler AS, Tissier F, Volante M, Matias-Guiu X, Smid M, Favier J, Rapizzi E, Libe R, Currás-Freixes M, Aydin S, Huynh T, Lichtenauer U, van Berkel A, Canu L, Domingues R, Clifton-Bligh RJ, Bialas M, Vikkula M, Baretton G, Papotti M, Nesi G, Badoual C, Pacak K, Eisenhofer G, Timmers HJ, Beuschlein F, Bertherat J, Mannelli M, Robledo M, Gimenez-Roqueplo A-P, Dinjens WNM, Korpershoek E, de Krijger RR (2015) SDHB/SDHA immunohistochemistry in pheochromocytomas and paragangliomas: a multicenter interobserver variation analysis using virtual microscopy: a Multinational Study of the European Network for the Study of Adrenal Tumors (ENS@T). Mod Pathol 28:807–821CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Olivera Casar-Borota
    • 1
    • 2
    • 3
    Email author
  • Kristin Astrid Berland Øystese
    • 4
    • 5
  • Magnus Sundström
    • 1
    • 2
  • Linea Melchior
    • 6
  • Vera Popovic
    • 7
  1. 1.Department of Immunology, Genetics and PathologyUppsala University, Rudbeck LaboratoryUppsalaSweden
  2. 2.Department of Clinical Pathology and CytologyUppsala University Hospital, Rudbeck LaboratoryUppsalaSweden
  3. 3.Department of PathologyOslo University HospitalOsloNorway
  4. 4.Department of Specialised EndocrinologyOslo University HospitalOsloNorway
  5. 5.Faculty of MedicineUniversity of OsloOsloNorway
  6. 6.Department of PathologyRigshospitalet Copenhagen University HospitalCopenhagenDenmark
  7. 7.Medical FacultyUniversity of BelgradeBelgradeSerbia

Personalised recommendations