Molecular diagnosis of pituitary adenoma
predisposition caused by aryl hydrocarbon
receptor-interacting protein gene mutations
Marianthi Georgitsia, Anniina Raitilaa, Auli Karhua, Karoliina Tuppurainenb, Markus J. Ma ¨kinenb, Outi Vierimaac,
Ralf Paschked, Wolfgang Saegere, Rob B. van der Luijtf, Timo Saneg, Mercedes Robledoh, Ernesto De Menisi,
Robert J. Weilj, Anna Wasikk, Grzegorz Zielinskil, Olga Lucewiczm, Jan Lubinskik,m, Virpi Launonena,
Pia Vahteristoa, and Lauri A. Aaltonena,n
aDepartment of Medical Genetics, Molecular and Cancer Biology Research Program, University of Helsinki, P.O. Box 63, 00014, Helsinki, Finland;
bDepartment of Pathology, University of Oulu, P.O. Box 5000, 90014, Oulu, Finland;cDepartment of Clinical Genetics, Oulu University Hospital, P.O. Box 60,
90029, Oulu, Finland;dMedical Department III, Leipzig University, Ph-Rosenthal-Street 27, 04103 Leipzig, Germany;eInstitute of Pathology,
Marienkrankenhaus, Alfredstrasse 9, 22087 Hamburg, Germany;fDepartment of Medical Genetics, University Medical Centre Utrecht, P.O. Box 85090, 3508
GA, Utrecht, The Netherlands;gDepartment of Endocrinology, Helsinki University Central Hospital, P.O. Box 340, 00029, Helsinki, Finland;hHereditary
Endocrine Cancer Group, Human Cancer Genetics Programme, Spanish National Cancer Center (CNIO), Melchor Ferna ´ndez Almagro 3, 28029 Madrid, Spain;
iDepartment of Internal Medicine, General Hospital, Piazza Ospedale 1, 31100 Treviso, Italy;jBrain Tumor Institute and Department of Neurosurgery,
Cleveland Clinic, 9500 Euclid Avenue, Cleveland, OH 44195;kDepartment of Cell Biology, Nencki Institute of Experimental Biology, Pasteura 3, 02093,
Warsaw, Poland;lDepartment of Neurosurgery, Military Institute of the Health Services, Szaserow 128, 00909, Warsaw, Poland; andmDepartment of
Genetics and Pathology, International Hereditary Cancer Center, Pomeranian Medical University, Polabska 4, 70115, Szczecin, Poland
Communicated by Bert Vogelstein, The Sidney Kimmel Comprehensive Cancer Center at Johns Hopkins, Baltimore, MD, January 2, 2007 (received for review
November 28, 2006)
Pituitary adenomas are common neoplasms of the anterior pituitary
gland. Germ-line mutations in the aryl hydrocarbon receptor-inter-
acting protein (AIP) gene cause pituitary adenoma predisposition
In this population, a founder mutation explained a significant pro-
portion of all acromegaly cases. Typically, PAP patients were of a
young age at diagnosis but did not display a strong family history of
pituitary adenomas. To evaluate the role of AIP in pituitary adenoma
susceptibility in other populations and to gain insight into patient
selection for molecular screening of the condition, we investigated
the possible contribution of AIP mutations in pituitary tumorigenesis
were investigated by AIP sequencing: young acromegaly patients,
unselected acromegaly patients, unselected pituitary adenoma pa-
tients, and endocrine neoplasia-predisposition patients who were
negative for MEN1 mutations. Nine AIP mutations were identified.
Because many of the patients displayed no family history of pituitary
of AIP immunohistochemistry (IHC) as a prescreening tool was tested
in 50 adenomas: 12 AIP mutation-positive versus 38 mutation-nega-
tive pituitary tumors. AIP IHC staining levels proved to be a useful
predictor of AIP status, with 75% sensitivity and 95% specificity for
germ-line mutations. AIP contributes to PAP in all studied popula-
tions. AIP IHC, followed by genetic counseling and possible AIP
mutation analysis in IHC-negative cases, a procedure similar to the
diagnostics of the Lynch syndrome, appears feasible in identification
immunohistochemistry ? growth hormone/prolacting–secreting adenomas ?
?15% of intracranial tumors (1). Approximately two-thirds
produce pituitary hormones in excess; among these, prolactin
(PRL)- and growth hormone (GH)-oversecreting adenomas are
the most common. GH-secreting adenomas cause acromegaly
and gigantism. Less common are adrenocorticotropin hormone
(ACTH)-secreting adenomas, causing Cushing’s disease. The
remaining one-third of pituitary adenomas is endocrinologically
silent, known as nonfunctioning pituitary adenomas, and cause
symptoms or signs due to tumor growth (1–3). Pituitary adeno-
mas are components of rare, well established syndromes, such as
ituitary adenomas are common, benign, monoclonal neo-
plasms of the anterior pituitary gland. They account for
multiple endocrine neoplasia type 1 (MEN1) and Carney com-
plex (CNC) (4, 5). Recent data suggest that a genetic predispo-
sition to pituitary tumors is less rare than thought and that genes
other than those for MEN1 and CNC are also involved (5–7).
Recently, we showed that germ-line mutations of the aryl
hydrocarbon receptor-interacting protein (AIP) gene cause pi-
tuitary adenoma predisposition (PAP) (8). A nonsense muta-
mutation segregated perfectly with the GH phenotype and was
also present in three prolactinoma patients. In addition, a
nonsense mutation, p.R304X, was found in two Italian siblings
with GH-secreting adenomas. In a population-based series from
Northern Finland, AIP mutations accounted for 16% of all
patients diagnosed with pituitary adenomas secreting GH and
for 40% of patients younger than 35 years of age. Typically, PAP
patients were of a young age at disease onset and did not display
a strong family history of pituitary adenomas. Loss of the normal
allele was detected in eight of eight pituitary adenomas; AIP is
likely to act as a tumor suppressor gene (8).
AIP encodes a protein of 330 aa. The protein contains an
FKBP-homology domain, and three tetratricopeptide (TPR)
repeats. AIP forms interactions with the aryl hydrocarbon
receptor (AHR, also known as dioxin receptor), two HSP90
proteins, PDE4A5, PPAR?, and survivin (9–12).
In our first study, for gene identification purposes, we focused
on a defined, homogeneous population (8). To gain insight into
clinical features and approaches to diagnose the condition, it was
relevant to examine the contribution of germ-line AIP mutations
in other patient materials as well. Here we sequenced the whole
AIP coding region in a large, heterogeneous collection of 460
V.L., P.V., and L.A.A. designed research; M.G., A.R., K.T., and M.J.M. performed research;
R.P., W.S., R.B.v.d.L., T.S., M.R., E.D.M., R.J.W., A.W., G.Z., O.L., and J.L. contributed new
reagents/analytic tools; M.G., A.R., A.K., K.T., M.J.M., V.L., P.V., and L.A.A. analyzed data;
and M.G., A.R., A.K., E.D.M., R.J.W., V.L., P.V., and L.A.A. wrote the paper.
The authors declare no conflict of interest.
protein; GH, growth hormone; IHC, immunohistochemistry; MEN1, multiple endocrine
neoplasia type 1; PAP, pituitary adenoma predisposition; PRL, prolactin.
database (accession nos. EF203234–EF203240).
nTo whom correspondence should be addressed. E-mail: firstname.lastname@example.org.
© 2007 by The National Academy of Sciences of the USA
www.pnas.org?cgi?doi?10.1073?pnas.0700004104 PNAS ?
March 6, 2007 ?
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pituitary adenoma patients and patients from families with
MEN1 features and who were derived from different popula-
tions in Europe and the United States. In addition, because the
genetic evidence suggests that many AIP-associated pituitary
adenomas are null for AIP protein, we tested whether negative
staining in AIP immunohistochemistry (IHC) in pituitary ade-
nomas would be a useful marker for PAP.
Mutation Analysis. Nine presumably pathogenic mutations were
identified. These mutations, and the features of the respective
patients, are depicted in Table 1 and Fig. 1. It is typically
challenging to robustly evaluate the nature of missense changes
in hereditary predisposition. Here, if a missense change was not
seen in controls and was associated with a phenotype strongly
suggestive of PAP, it was presumed pathogenic. Other changes
were presumed to be neutral. It is clear that this subdivision is
preliminary and should be interpreted with some caution. The
results are reviewed and the details of the missense changes are
Young acromegaly patients. In the German samples, two AIP
mutations were identified (2 of 27, 7.4%; see Table 1). In
addition, a heterozygous intronic change, IVS1-18C3T, was
identified in one sample but was not predicted to have an effect
on splicing as tested in silico. This same intronic change was also
detected in 1 of 107 Centre d’E´tude du Polymorphism Humain
controls (1%) and 1 of 96 Caucasian U.K. controls (1%),
suggesting that the variant is a polymorphism.
From the Helsinki University Central Hospital (HUCH)
patient cohort, 36 Finnish patients were analyzed. The Finnish
germ-line founder mutation was identified in two (5.5%; see
Table 1) patients.
Unselected acromegaly patients. Among the 71 Italian sporadic
acromegaly patients, one heterozygous germ-line missense
change was found: p.R16H (c.47G3A, resulting in the substi-
tution of arginine at position 16 by histidine). Loss-of-
heterozygosity analysis from this individual’s pituitary tumor
tissue was negative. p.R16H was absent in 181 Caucasian U.K.,
52 Italian, and 209 Finnish controls. The change was found in 1
of 90 healthy German controls (1%), 1 unselected pituitary
adenoma patient from the United States, and in 3 Polish
unselected pituitary adenoma patients (see below), suggesting
that this change may be a neutral polymorphism.
Table 1. AIP mutations identified in pituitary adenoma patients from the European and North American populations
(%) Clinical data
Germanyc.66-71delAGGAGAExon1 1 of 27 (3.7)Acromegaly-
0 of 532
c.878-879AG3GT (p.E293G) and
c.40C3T (p.Q14X )
Exon6 1of 27 (3.7) YesF 29†
0 of 255
FinlandExon1 2 of 36 (5.5) NAM 36No 0 of 532
——— NAF 41 No 0 of 532
Italy—— 0 of 71——————
U.S. IVS2-1G3C Intron 2 1 of 113 (0.9) Acromegaly-
NAM 20 No0 of 202
1 of 113 (0.9)
1 of 122 (0.8)
0 of 201
0 of 255
Spainc.542delTExon4 1 of 55 (1.8) Acromegaly–
0 of 203
The Netherlandsc.896C3T (p.A299V)Exon6 1 of 36 (2.8)NAF 160 of 255
ACTHoma, ACTH-secreting adenoma; F, female; GHoma, GH-secreting adenoma; M, male; NA, not available.
*Only putative pathogenic changes are depicted.
†Age at time of operation; age at time of diagnosis is not known.
identified in this study. The locations of the FKBP-homology region and the
three tetratricopeptide repeats (TPRs) are indicated by colored boxes. AHR
and HSP90 interaction regions are indicated by black lines.
Diagram of AIP displaying the presumably pathogenic mutations
www.pnas.org?cgi?doi?10.1073?pnas.0700004104Georgitsi et al.
Unselected pituitary adenoma patients. AIP mutation analysis per-
formed in 113 unselected pituitary adenoma patients from the
Table 1) and two heterozygous missense changes that are likely to
be polymorphisms. A heterozygous c.906G3A, resulting in the
silent p.V302V change in exon 6, was found in three cases. When
p.V302V was tested in silico, the prediction programs showed no
history of pituitary adenomas. Loss-of-heterozygosity analysis was
possible from tumor DNA samples of 2/3 individuals, and showed
retention of the wild type allele. p.V302V was not detected in 109
52 Italian controls. Finally, the previously seen missense change
of Polish descent. Tumor DNA sequence did not show loss of
In 122 unselected pituitary adenoma patients from Poland,
three different germ-line heterozygous missense changes were
detected, of which one was considered disease associated (1 of
122, 0.8%; Table 1). In addition, the previously detected p.R16H
change was seen also in three Polish individuals all diagnosed
with Cushing’s disease. A heterozygous c.696G3C, resulting in
the silent p.P232P change in exon 5, was found in one patient
with Cushing’s disease. This change did not have any predicted
effect on splicing as tested in silico. p.P232P was absent in 108
Centre d’E´tude du Polymorphism Humain or 95 Caucasian U.K.
Patients counseled and examined for MEN1 with negative genetic testing
identified (1 of 55, 1.8%; see Table 1). Likewise, of the 36 Dutch
samples, an AIP germ-line mutation was identified in one
specimen (1 of 36, 2.8%; see Table 1).
AIP Immunohistochemical Staining. AIP immunoreaction was ob-
served in both the cytoplasm and the nucleus in normal adeno-
of 38) had preserved cytoplasmic and nuclear immunoreaction
against AIP (Fig. 2B), whereas most AIP-deficient adenomas (9
of 12) lacked both cytoplasmic and nuclear immunoreactivity
against AIP (Fisher’s Exact test, P ? 0.000004). In tumor tissues,
leukocytes served as internal positive controls (Fig. 2 C and D).
AIP IHC had 75% sensitivity and 95% specificity for truncating
AIP germ-line mutations.
In our original study, we evaluated the contribution of AIP in a
population-based material of acromegaly patients, diagnosed
between 1980 and 1999 in the Oulu region of Northern Finland.
In this isolated population, two germ-line AIP mutations
(p.Q14X and IVS3-1G3A) accounted for 16% of all patients
diagnosed with pituitary adenomas secreting GH and for 40% of
patients that were younger than 35 years at the age of diagnosis
(8). In the current study, we examined the role of AIP in more
heterogeneous patient groups to provide clues to clinical and
molecular identification of PAP.
The analysis of 71 Italian acromegaly patients systematically
collected from the Treviso region revealed no significant find-
ings, but two siblings (2 of 73, 2.7%) belonging to this same
sample collection were shown to display a truncating AIP
mutation in our previous study (8).
In both of the two patient sets with acromegaly that presented
at a young age, the Helsinki and Leipzig regions, two PAP
patients were detected; altogether, 4 of 63 (6.3%) patients
Two putative AIP mutations were found among unselected
pituitary adenoma cases from the United States. IVS2-1G3C
splice site mutation was detected in a patient diagnosed with
acromegaly at the age of 20 years. c.824–825insA insertion, also
causing a premature stop codon, was seen in a patient diagnosed
with GH-secreting adenoma at the age of 8 years. These two
cases account for 1.8% of the unselected pituitary adenoma
cases in this series. The number of acromegaly patients in the
sample set was only 13. Although the numbers are very small, it
is noteworthy that the contribution of PAP in acromegaly in this
consecutive U.S. series was similar to that seen in Northern
Finland (2 of 13, 15%).
cells, whereas AIP heterozygous peripheral blood leucocytes display positive immunoreaction, indicated by black arrows.
AIP IHC. (A) AIP immunoreaction is observed in both the cytoplasm and the nucleus in normal adenohypophysis. (B) AIP expression in AIP-proficient
Georgitsi et al.
March 6, 2007 ?
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p.R304Q change was seen in one Polish unselected pituitary
would be previously unrecognized in Cushing’s disease. We have
A.R., A.K., E.D.M., V.L., P.V., and L.A.A., unpublished data)
but in none of the healthy controls, which strongly supports the
notion that the change is pathogenic. Interestingly, p.R304Q
locates on the AHR-binding region (13, 14) and possibly affects
the interaction of AIP with AHR.
Approximately 10% of cases clinically suggestive of MEN1 do
not seem to harbor germ-line MEN1 mutations (4, 15–17). Thus,
we examined whether AIP is involved in such cases of unex-
plained endocrine neoplasia susceptibility. Two mutation-
positive cases were found (2 of 91, 2%). The phenotypes of these
two patients were in line with the above findings; both the
Spanish and the Dutch patient were diagnosed with acromegaly
at an early age: 18 and 16 years, respectively. The Spanish patient
also had a positive family history, with two maternal uncles being
diagnosed with acromegaly.
These data firmly confirm that AIP is directly implicated in the
molecular pathogenesis of pituitary tumors, particularly of the
GH/PRL lineage. In the future, it will be of interest to examine
what the contribution of de novo mutations in PAP is. In the
current work, the relevant additional samples were not available.
The prevalence of AIP germ-line mutations varies in different
clinical settings. In unselected sporadic pituitary tumors, the
overall prevalence seems to be low: 2 of 113 and 1 of 122 from
our U.S. and Poland cases, respectively. Also, none of the
previously reported p.Q14X, p.R304X, and IVS3-1G3A mu-
tations were found in a recently published U.S. series (18). On
the contrary, AIP mutations are enriched in patients of a very
although some of the PAP patients display neither of these
features. Selection of patients for genetic counseling and possi-
ble genetic testing for PAP appeared challenging.
To help simplify selection, we tested AIP IHC in 50 pituitary
mutations are truncating and because the condition is typically
associated with loss of the wild-type allele in tumors, a strategy
similar to that used to screen patients for hereditary nonpolyposis
screening of tumors for loss of the predisposition gene product,
displaying negative IHC for AIP, followed by cascade screening in
family members to identify individuals at risk. Because pituitary
adenomas are examined immunohistochemically as a routine di-
agnostic practice, this screening method appears to be feasible.
Indeed, we found that negative AIP IHC staining is a strong
predictor of PAP. Two cases with a negative AIP mutation analysis
detected by sequencing) germ-line mutations or somatic loss of
AIP, although technical problems are also a possible explanation.
40 and 47 years of age, and, thus, PAP is possible. Similarly, in the
three mutation-positive cases displaying positive AIP IHC, the
unexpected IHC finding could have been due to technical difficul-
ties such as unspecific staining or missense-type second hits en-
abling production of nonfunctional yet immunoreactive AIP pro-
ability to screen for AIP alterations by IHC will further improve.
in the near future, IHC screening will remain useful, because direct
DNA testing for AIP germ-line mutations would require prior
genetic counseling. Genetic counseling demands resources and
needs to be reserved for those pituitary adenoma cases that display
features of hereditary susceptibility.
The annual incidence of newly diagnosed cases of acromegaly
ranges from 2.8 to ?4 per million, with prevalences ranging from
34 to 120 cases per million (20). Although these observations
suggest that ?1,000 new cases will be diagnosed annually in the
United States, the insidious nature of acromegaly and the frequent
delays in diagnosis in this disease have led to estimates that
majority of patients present with a macroadenoma, and younger
patients frequently have larger, more invasive tumors with poorer
outcomes, the potential for prolonged biochemical remission with
any single modality is diminished (22, 23). In patients with more
advanced disease, successful therapy eliminates or resolves all
manifestations of the disease in a minority, and diminished quality-
related primarily to chronic cardiovascular disease, is a function of
biochemical control: risk of mortality may be as high as 3.5-fold
greater in patients with persistent disease compared with those in
remission (22, 23, 25–28). Although molecular diagnosis of PAP in
unselected pituitary adenomas would be first performed in a
research setting, thousands of paraffin-embedded somatotropi-
noma samples in the United States alone are available for pre-
screening of PAP by AIP IHC, pending consent from the patients.
does not seem to be influenced by diagnosed AIP mutation
positivity, offering genetic counseling and predictive testing to
family members provides a powerful tool for prevention of mor-
bidity in at-risk individuals. The great majority of AIP-related
tumors are GH- and/or PRL-secreting adenomas, and the clinical
diagnosis of acromegaly at onset is difficult. Therefore, we suggest
that asymptomatic relatives testing positive for an AIP mutation
should undergo annual PRL and IGF1 monitoring (29), as sug-
gested for MEN1 carriers (30). Finally, we highlight the particular
for acromegaly or with early onset of the tumor.
Materials and Methods
Study Subjects. The study was approved by the appropriate ethics
review committees. Appropriate informed consent was obtained
from all subjects.
Young acromegaly patients. DNA extracted from paraffin-
embedded tumor tissue was available from 27 patients with
acromegaly from the German pituitary tumor register, Institute
of Pathology, Marienkrankenhaus Hamburg. The search was
conducted for entries during the last 3 years for patients younger
than 40 years old at the time of surgery.
DNA derived from blood was available from 36 Finnish acro-
megaly patients who were ?45 years old and originally diagnosed
and treated at the Department of Endocrinology, Helsinki Uni-
versity Central Hospital (HUCH). This cohort represented 57.1%
of all young (?45 years) acromegaly patients diagnosed at HUCH
between the years 1980–2005.
Unselected acromegaly patients. Blood-extracted DNA samples from
71 unselected Italian acromegaly patients who were referred to
Treviso General Hospital were available. Age at diagnosis ranged
between 23 and 90 years, with a mean age of 45 years.
Unselected pituitary adenoma patients. Altogether, 113 samples col-
lected consecutively from patients undergoing resection of a pitu-
itary tumor at the Cleveland Clinic were analyzed. DNA was
isolated from either blood or tumor tissue. Age at diagnosis ranged
between 8 and 87 years, with a mean age of 52 years. Of these 113
patients, all underwent biochemical and immunohistochemically
due to GH-secreting adenomas, 11 with hyperprolactinemia due to
PRL-secreting adenomas. The remaining 76 patients had a non-
functioning pituitary adenoma. None of the patients had a family
history of pituitary tumors.
Blood-extracted DNA samples from 122 unselected Polish pitu-
itary adenoma patients were collected at the International Hered-
itary Cancer Center, Pomeranian Medical University, in Szczecin,
www.pnas.org?cgi?doi?10.1073?pnas.0700004104 Georgitsi et al.
Poland. Of these, 74 patients were diagnosed with Cushing’s Download full-text
disease, 30 with acromegaly, and 18 with pituitary adenomas of
various types. Age of onset ranged between 8 and 67 years, with a
mean age of 39 years. Age of onset was not known for 21 cases.
Patients counseled and examined for MEN1 with negative genetic testing
Patients had been referred to the DNA Diagnostics Laboratory
(Department of Medical Genetics, University Medical Centre
the period of 2004–2006. Patients suspected for MEN1 were
defined as those with at least three of the following five lesions:
hyperparathyroidism/parathyroid tumors, pancreatic endocrine tu-
mors, pituitary adenomas, adrenal gland tumors, and/or neuroen-
docrine carcinoid tumors. The patients fulfilled the following
criteria: young age at onset (?35 years) of any of the five MEN1-
related lesions and/or multiple MEN1-related lesions in a single
organ or two distinct organs, and at least one first-degree relative
in whom at least one target organ was affected. Age at tumor
diagnosis ranged between 15 and 81 years, with a mean age of 50
Another set consisted of individuals suspected for MEN1 and
referred to the Spanish National Cancer Center during the period
of 1997–2006. Blood-extracted DNA samples from 55 unselected
and consecutive MEN1-negative patients were available for AIP
with a mean age of 50 years. Information was not available for two
Control Samples. DNAfromunrelated,anonymous,individualswas
used as control samples: 110 Caucasian Centre d’E´tude du Poly-
morphism Humain individuals, 288 Caucasians from the U.K.
(Human Random Control DNA Panels, Porton Down, Salisbury,
Wiltshire, U.K.), 209 Finns, 90 Germans, and 52 Italians.
IHC Samples. AIP protein expression was analyzed in 50 pituitary
adenomas. Twelve tumors were from AIP mutation-positive indi-
viduals (nine cases with p.Q14X, and one case with IVS2-1G3C,
c.824–825insA, and IVS3-1G3A, respectively) including 10 so-
matotropinomas and two prolactinomas. Thirty-eight mutation-
negative adenomas included 32 somatotropinomas, five prolacti-
nomas, and one GH- and PRL-negative adenoma.
Mutation Analysis. Mutation analysis was performed by direct
sequencing of genomic DNA. The whole coding region of AIP was
sequenced, as well as flanking intronic sequences and 5? and 3?
untranslated regions. PCR protocols and primer sequences have
been described by Vierimaa et al. (8) and are available on request.
DNA sequencing was performed using Big Dye 3.1 termination
chemistry on an ABI3730 DNA sequencer (Applied Biosystems,
Foster City, CA).
In Silico Analysis. The potential effects on splicing of the detected
intronic and silent changes were predicted in silico by computa-
tional methods by using NetGene2, Alternative Splice Site Predic-
tor (ASSP), and SpliceScan programs (31–34).
AIP IHC. Five-micrometer-thick sections were cut from the paraffin
blocks. After deparaffinization and rehydration, sections were
pretreated in either 0.01 M citrate (pH 6.0) buffer in a microwave
oven at 800 W for 2 min and at 300 W for 10 min or in 0.01 M
and at 300 W for 15 min. AIP was detected in tumors by using AIP
antibody (AIP SP5213P; Acris Antibodies, Hiddenhausen, Ger-
many) at a 1:4,000 dilution for 30 min. Positive antibody reaction
was detected with diaminobenzidine (DAKO, Copenhagen, Den-
mark) with hematoxylin counterstain.
for help with the in silico analysis; S. Marttinen, I. L. Svedberg, I. Vuoristo,
P. Hannuksela, M. Aho, and R. Vuento for their excellent technical
assistance; P. Ellonen for providing sequencing facilities and service; and
D. K. Luedecke (University of Hamburg, Germany) for providing material
and clinical data. This study was supported by Academy of Finland Grants
213183 (to V.L.) and 212901 (to P.V.), the Center of Excellence in
Translational Genome-Scale Biology, the Sigrid Juse ´lius Foundation, the
Cancer Society of Finland, Association for International Cancer Research
Grant 05-001 (to A.K.), a Jalmari and Rauha Ahokas Foundation research
grant (to M.G.), a Bodossaki Foundation postgraduate scholarship (to
M.G.), the Melvin Burkhardt Chair in Neurosurgical Oncology, and the
Karen Colina Wilson Research Endowment within the Brain Tumor
Institute at the Cleveland Clinic Foundation.
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