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    <TD>Child's Nervous System</TD></TR>
  <TR>
    <TD>=A9&nbsp;The Author(s)&nbsp;2008</TD></TR>
  <TR>
    <TD>10.1007/s00381-007-0578-0</TD></TR></TBODY></TABLE><!--Begin =
Abstract-->
<H2 class=3Drubric>Case Report</H2>
<DIV class=3DHeading1><A name=3Dtitle></A>Secondary meningioma in a =
long-term=20
survivor of atypical teratoid/rhabdoid tumour with a germline =
<I>INI1</I>=20
mutation </DIV>
<P class=3DAuthorGroup>A.&nbsp;C.&nbsp;J.&nbsp;Ammerlaan<SUP>1</SUP>,=20
M.&nbsp;P.&nbsp;W.&nbsp;A.&nbsp;Houben<SUP>2</SUP>,=20
C.&nbsp;C.&nbsp;Tijssen<SUP>2</SUP>, P.&nbsp;Wesseling<SUP>3</SUP> and=20
T.&nbsp;J.&nbsp;M.&nbsp;Hulsebos<SUP>1&nbsp;<A=20
href=3D"http://www.springerlink.com/content/0428100127358214/fulltext.htm=
l#ContactOfAuthor5"><IMG=20
alt=3D"Contact Information"=20
src=3D"http://www.springerlink.com/content/0428100127358214/contact.gif" =

border=3D0></A></SUP></P>
<TABLE>
  <TBODY>
  <TR vAlign=3Dtop>
    <TD><SPAN class=3DAffiliation><A =
name=3DAff1></A>(1)&nbsp;</SPAN></TD>
    <TD><SPAN class=3DAffiliation>Department of Neurogenetics, Academic =
Medical=20
      Center, Meibergdreef 9, 1105 AZ&nbsp;Amsterdam, The=20
  Netherlands</SPAN></TD></TR></TBODY></TABLE>
<TABLE>
  <TBODY>
  <TR vAlign=3Dtop>
    <TD><SPAN class=3DAffiliation><A =
name=3DAff2></A>(2)&nbsp;</SPAN></TD>
    <TD><SPAN class=3DAffiliation>Department of Neurology, St. Elisabeth =

      Hospital, Tilburg, The =
Netherlands</SPAN></TD></TR></TBODY></TABLE>
<TABLE>
  <TBODY>
  <TR vAlign=3Dtop>
    <TD><SPAN class=3DAffiliation><A =
name=3DAff3></A>(3)&nbsp;</SPAN></TD>
    <TD><SPAN class=3DAffiliation>Department of Pathology, Nijmegen =
Center for=20
      Molecular Life Sciences, Radboud University Nijmegen Medical =
Centre,=20
      Nijmegen, The Netherlands</SPAN></TD></TR></TBODY></TABLE>
<P><A name=3DContactOfAuthor5></A></P>
<TABLE class=3DContact>
  <TBODY>
  <TR>
    <TD vAlign=3Dtop><IMG alt=3D"Contact Information"=20
      =
src=3D"http://www.springerlink.com/content/0428100127358214/contact.gif" =

      border=3D0></TD>
    =
<TD><STRONG>T.&nbsp;</STRONG><STRONG>J.&nbsp;</STRONG><STRONG>M.&nbsp;</S=
TRONG><STRONG>Hulsebos</STRONG><STRONG></STRONG><BR><STRONG>Email:=20
      </STRONG><A=20
      =
href=3D"mailto:t.j.hulsebos@amc.uva.nl">t.j.hulsebos@amc.uva.nl</A></TD><=
/TR></TBODY></TABLE>
<P class=3DAffiliation><STRONG>Received:=20
</STRONG>21&nbsp;December&nbsp;2007&nbsp;&nbsp;<STRONG>Published online: =

</STRONG>31&nbsp;January&nbsp;2008 </P>
<DIV class=3DAbstract><A name=3DAbs1></A><SPAN =
class=3DAbstractHeading>Abstract</SPAN>
<DIV class=3DAbstractSection>
<DIV class=3D""><SPAN class=3DAbstractSectionHeading><A=20
name=3DASec1></A>Objective&nbsp;&nbsp;</SPAN>We report on a patient who =
developed=20
a meningioma more than two decades after removal at a young age of an =
atypical=20
teratoid/rhabdoid tumour (AT/RT), which was due to a germline =
<I>INI1</I>=20
mutation, and radio- and chemotherapy. </DIV></DIV>
<DIV class=3DAbstractSection>
<DIV class=3D""><SPAN class=3DAbstractSectionHeading><A =
name=3DASec2></A>Materials and=20
methods&nbsp;&nbsp;</SPAN>We present genetic evidence that the =
meningioma is not=20
a recurrence or metastasis of the AT/RT and not due to the <I>INI1</I> =
mutation,=20
but is a radiation-induced tumour. </DIV></DIV>
<DIV class=3DAbstractSection>
<DIV class=3D""><SPAN class=3DAbstractSectionHeading><A=20
name=3DASec3></A>Conclusion&nbsp;&nbsp;</SPAN>This is the first case =
illustrating=20
that improved survival of young patients with an AT/RT after aggressive=20
treatment may be gained at the cost of an increased risk for the =
development of=20
radiation-induced, non-<I>INI1</I>-related tumours. </DIV></DIV></DIV>
<P class=3DKeyword><SPAN=20
class=3DKeywordHeading>Keywords&nbsp;&nbsp;</SPAN>Meningioma&nbsp;-&nbsp;=
Radiation=20
therapy&nbsp;-&nbsp;Atypical teratoid/rhabdoid =
tumour&nbsp;-&nbsp;Genetic=20
predisposition&nbsp;-&nbsp; <I>INI1</I> </P>
<DIV class=3D""><A name=3DSec1></A>
<HR>

<DIV class=3Dheading2>Introduction</DIV>
<P class=3D"">We previously reported on a family in which four cousins =
developed=20
an atypical teratoid/rhabdoid tumour (AT/RT) at young age, due to =
inheritance of=20
a germline G&gt;A mutation in the donor splice site of exon 4 of the =
tumour=20
suppressor gene <I>INI1</I> [<CITE><A=20
href=3D"http://www.springerlink.com/content/0428100127358214/fulltext.htm=
l#CR2">2</A></CITE>].=20
More than 20&nbsp;years after resection of the AT/RT, the oldest of the =
four=20
cousins developed an intracranial meningioma and, after an additional =
year, a=20
myoepithelioma of the lip. In addition to the constitutional <I>INI1</I> =

mutation and loss of the normal <I>INI1</I> allele, we detected an =
identical=20
rearrangement in the <I>NF2</I> gene, not affecting its coding regions, =
in the=20
AT/RT and myoepithelioma of the patient and concluded that they both =
originated=20
from a common precursor cell. However, the rearrangement in <I>NF2</I> =
and loss=20
of the normal <I>INI1</I> allele could not be demonstrated in the =
meningioma.=20
Despite this, the development of the meningioma may be the consequence =
of the=20
presence of the constitutional <I>INI1</I> mutation, because mutations =
in this=20
gene have been identified in meningiomas [<CITE><A=20
href=3D"http://www.springerlink.com/content/0428100127358214/fulltext.htm=
l#CR8">8</A></CITE>].=20
Alternatively, the meningioma could be induced by the radiation therapy, =
which=20
our patient received after removal of the AT/RT and which is a =
well-known=20
consequence of this treatment [<CITE><A=20
href=3D"http://www.springerlink.com/content/0428100127358214/fulltext.htm=
l#CR1">1</A></CITE>].=20
We investigated the meningioma for the presence of genetic changes =
reported in=20
radiation-induced meningiomas and performed a comparative loss of =
heterozygosity=20
(LOH) analysis to demonstrate that such radiation-induced genetic =
changes are=20
absent in the AT/RT and myoepithelioma of our patient. </P></DIV>
<DIV class=3D""><A name=3DSec2></A>
<HR>

<DIV class=3Dheading2>Materials and methods</DIV>
<DIV class=3D""><A name=3DSec3></A>
<DIV class=3DHeading3>Tumour samples and DNA isolation</DIV>
<P class=3D"">DNA from the AT/RT and the myoepithelioma was isolated =
from=20
formalin-fixed and paraffin-embedded tumour material, and DNA from the=20
meningioma was isolated from a fresh frozen tumour sample using =
commercially=20
available kits (Qiagen, Venlo, The Netherlands). </P></DIV>
<DIV class=3D""><A name=3DSec4></A>
<DIV class=3DHeading3>SNP analysis</DIV>
<P class=3D"">A genome-wide single nucleotide polymorphism (SNP) =
analysis of=20
normal and meningioma DNA was conducted with the Affymetrix GeneChip =
Human=20
Mapping 10K Array Xba 142 2.0. The array contains 10,204 SNPs =
distributed across=20
the genome. The GeneChips were processed by an authorised Affymetrix =
Service=20
Provider using standard Affymetrix protocols (Service XS, Leiden, The=20
Netherlands). LOH regions were identified by comparing the SNP profiles =
of=20
normal and meningioma DNA and scoring heterozygous SNP markers (AB) in =
the=20
normal DNA that were reduced to homozygosity (AA or BB) in the =
meningioma DNA.=20
Intervals were determined on the basis of locations of SNP markers in =
the UCSC=20
Genome Browser, March 2006 Build, at <A=20
href=3D"http://genome.ucsc.edu/">http://genome.ucsc.edu/</A>. </P></DIV>
<DIV class=3D""><A name=3DSec5></A>
<DIV class=3DHeading3>LOH analysis</DIV>
<P class=3D"">Loss of heterozygosity (LOH) analysis was performed with=20
microsatellite markers as described previously [<CITE><A=20
href=3D"http://www.springerlink.com/content/0428100127358214/fulltext.htm=
l#CR3">3</A></CITE>]=20
using primer sequences and conditions for polymerase chain reaction =
given by The=20
Genome Database at <A =
href=3D"http://www.gdb.org/">http://www.gdb.org/</A>.=20
Because of rather extensive degradation of DNA extracted from the AT/RT, =
only=20
markers generating PCR fragments smaller than 150&nbsp;bp were used.=20
</P></DIV></DIV>
<DIV class=3D""><A name=3DSec6></A>
<HR>

<DIV class=3Dheading2>Case report</DIV>
<DIV class=3D""><A name=3DSec7></A>
<DIV class=3DHeading3>History</DIV>
<P class=3D"">Our patient was diagnosed with a tumour in the fourth =
ventricle and=20
right cerebellar hemisphere at the age of 4.5&nbsp;years. Resection of =
the=20
tumour was incomplete. The tumour was originally classified as =
anaplastic=20
ependymoma WHO grade III, but upon extensive genetic analysis and=20
histopathological re-evaluation, reclassified as AT/RT [<CITE><A=20
href=3D"http://www.springerlink.com/content/0428100127358214/fulltext.htm=
l#CR2">2</A></CITE>].=20
He received adjuvant chemotherapy (methotrexate, vincristine and =
prednisolone)=20
followed by craniospinal radiotherapy (3,300&nbsp;cGy in 22 doses) with =
a boost=20
of 2,100&nbsp;cGy on the location of the tumour. Fifteen years later, a =
brain CT=20
without contrast showed no signs of recurrent disease or of other new =
pathology.=20
Almost 6&nbsp;years later, 20.5&nbsp;years after initial diagnosis, he =
presented=20
again with bifrontal headaches, increasing ataxic gait disturbances and =
fatigue.=20
CT examination showed a large isodens space-occupying lesion in the left =

temporal region with oedema, midline shift and contrast enhancement. One =
year=20
after resection of this tumour, he developed a myoepithelioma of the =
upper lip.=20
The patient is alive at 29&nbsp;years of age. </P></DIV>
<DIV class=3D""><A name=3DSec8></A>
<DIV class=3DHeading3>Histological findings</DIV>
<P class=3D"">Histopathological examination of the left temporal tumour =
revealed=20
an atypical meningothelial meningioma WHO grade II with many vessels and =
up to=20
six mitoses per 2&nbsp;mm<SUP>2</SUP>. </P>
<DIV class=3DFormalPara>
<DIV class=3D""><SPAN style=3D"FONT-STYLE: italic; TEXT-DECORATION: =
none">SNP and=20
LOH analysis</SPAN>&nbsp;&nbsp; Radiation-induced meningiomas usually =
display a=20
complex karyotype [<CITE><A=20
href=3D"http://www.springerlink.com/content/0428100127358214/fulltext.htm=
l#CR1">1</A></CITE>].=20
To determine the genetic changes, we performed a genome-wide scan by SNP =

analysis of the meningioma and corresponding normal DNA. The identified=20
chromosomal regions displaying LOH are listed in Table&nbsp;<A=20
href=3D"http://www.springerlink.com/content/0428100127358214/fulltext.htm=
l#Tab1">1</A>.=20
Next, we tested several markers from within each of the five largest =
deleted=20
regions by conventional microsatellite analysis to determine their LOH =
status in=20
the meningioma and in the AT/RT and myoepithelioma. Presence of LOH in =
the=20
meningioma, but absence of LOH in AT/RT and myoepithelioma was found for =

<I>D1S199</I> in 1pter-p34.3 (Fig.&nbsp;<A=20
href=3D"http://www.springerlink.com/content/0428100127358214/fulltext.htm=
l#Fig1">1</A>a),=20
<I>D2S327</I> in 2pter-p12, <I>D3S3727</I> in 3p26.1-cen (Fig.&nbsp;<A=20
href=3D"http://www.springerlink.com/content/0428100127358214/fulltext.htm=
l#Fig1">1</A>b),=20
<I>D16S3020</I> in 16pter-p12.1 and <I>D17S1872</I> in 17qcen-q21.31=20
(Fig.&nbsp;<A=20
href=3D"http://www.springerlink.com/content/0428100127358214/fulltext.htm=
l#Fig1">1</A>c).=20
In contrast, as determined previously [<CITE><A=20
href=3D"http://www.springerlink.com/content/0428100127358214/fulltext.htm=
l#CR2">2</A></CITE>]=20
and exemplified by marker <I>D22S430</I> in Fig.&nbsp;<A=20
href=3D"http://www.springerlink.com/content/0428100127358214/fulltext.htm=
l#Fig1">1</A>d,=20
there was LOH for chromosome 22 markers in the AT/RT and myoepithelioma, =
but not=20
in the meningioma.=20
<DIV class=3DFigure><A name=3DFig1></A><IMG=20
alt=3DMediaObjects/381_2007_578_Fig1_HTML.gif=20
src=3D"http://www.springerlink.com/content/0428100127358214/MediaObjects/=
381_2007_578_Fig1_HTML.gif"></DIV>
<DIV class=3DCapt><SPAN class=3DCaptNr>Fig.&nbsp;1&nbsp;</SPAN>LOH =
analysis using=20
markers from chromosome regions <B>a</B> 1pter-p34.3 (<I>D1S199</I>), =
<B>b</B>=20
3p26.1-cen (<I>D3S3727</I>), <B>c</B> 17qcen-q21.31 (<I>D17S1872</I>) =
and=20
<B>d</B> 22q (<I>D22S430</I>) of AT/RT (<I>A</I>), meningioma (<I>M</I>) =
and=20
myoepithelioma (<I>My</I>) DNAs of the patient. <I>N</I> denotes normal =
DNA=20
derived from the patient=92s blood leucocytes. <I>Arrowhead</I> =
indicates position=20
of lost allele </DIV>
<HR>
<A name=3DTab1></A>
<DIV class=3DCapt><SPAN class=3DCaptNr>Table&nbsp;1&nbsp;</SPAN>LOH =
regions in the=20
meningioma, as determined by SNP analysis </DIV>
<TABLE border=3D1>
  <COLGROUP>
  <COL align=3Dleft>
  <COL align=3Dleft></COLGROUP>
  <THEAD>
  <TR class=3Dheader>
    <TH align=3Dleft>
      <P class=3D"">Chromosome region</P></TH>
    <TH align=3Dleft>
      <P class=3D"">Exact interval (Mb)</P></TH></TR></THEAD>
  <TBODY>
  <TR class=3Dnoclass>
    <TD align=3Dleft>
      <P class=3D"">1pter-p34.3</P></TD>
    <TD align=3Dchar char=3D"=96">
      <P class=3D"">0=9638.90</P></TD></TR>
  <TR class=3Dnoclass>
    <TD align=3Dleft>
      <P class=3D"">1p33-p33</P></TD>
    <TD align=3Dchar char=3D"=96">
      <P class=3D"">46.99=9651.09</P></TD></TR>
  <TR class=3Dnoclass>
    <TD align=3Dleft>
      <P class=3D"">1p32.2-1p31.3</P></TD>
    <TD align=3Dchar char=3D"=96">
      <P class=3D"">58.19=9665.50</P></TD></TR>
  <TR class=3Dnoclass>
    <TD align=3Dleft>
      <P class=3D"">2pter-p12</P></TD>
    <TD align=3Dchar char=3D"=96">
      <P class=3D"">0=9679.28</P></TD></TR>
  <TR class=3Dnoclass>
    <TD align=3Dleft>
      <P class=3D"">3p26.1-cen</P></TD>
    <TD align=3Dchar char=3D"=96">
      <P class=3D"">6.11=9688.84</P></TD></TR>
  <TR class=3Dnoclass>
    <TD align=3Dleft>
      <P class=3D"">7q36.1-qter</P></TD>
    <TD align=3Dchar char=3D"=96">
      <P class=3D"">150.04=96158.66</P></TD></TR>
  <TR class=3Dnoclass>
    <TD align=3Dleft>
      <P class=3D"">16pter-p12.1</P></TD>
    <TD align=3Dchar char=3D"=96">
      <P class=3D"">0=9622.75</P></TD></TR>
  <TR class=3Dnoclass>
    <TD align=3Dleft>
      <P class=3D"">16q23.1-q23.3</P></TD>
    <TD align=3Dchar char=3D"=96">
      <P class=3D"">73.46=9681.33</P></TD></TR>
  <TR class=3Dnoclass>
    <TD align=3Dleft>
      <P class=3D"">17cen-q21.31</P></TD>
    <TD align=3Dchar char=3D"=96">
      <P class=3D"">23.09=9640.35</P></TD></TR>
  <TR class=3Dnoclass>
    <TD align=3Dleft>
      <P class=3D"">19pter-p13.2</P></TD>
    <TD align=3Dchar char=3D"=96">
      <P =
class=3D"">0=968.83</P></TD></TR></TBODY></TABLE></DIV></DIV></DIV></DIV>=

<DIV class=3D""><A name=3DSec9></A>
<HR>

<DIV class=3Dheading2>Discussion</DIV>
<P class=3D"">In a previous study, we provided evidence for the =
involvement of=20
<I>INI1</I>, by constitutional mutation and independent somatic loss, in =
the=20
development of both the AT/RT and the myoepithelioma of our patient. =
Moreover,=20
we could demonstrate that these tumours originated from an identical =
precursor=20
cell [<CITE><A=20
href=3D"http://www.springerlink.com/content/0428100127358214/fulltext.htm=
l#CR2">2</A></CITE>].=20
Although involvement of <I>INI1</I> has been described in sporadic =
meningiomas=20
[<CITE><A=20
href=3D"http://www.springerlink.com/content/0428100127358214/fulltext.htm=
l#CR8">8</A></CITE>],=20
until now meningiomas have not been reported to occur as a consequence =
of the=20
inheritance of an <I>INI1</I> mutation. However, this might be due to =
the=20
extreme rarity of families with an inherited <I>INI1</I> mutation, with =
only=20
four published cases so far [<CITE><A=20
href=3D"http://www.springerlink.com/content/0428100127358214/fulltext.htm=
l#CR2">2</A></CITE>,=20
<CITE><A=20
href=3D"http://www.springerlink.com/content/0428100127358214/fulltext.htm=
l#CR5">5</A></CITE>,=20
<CITE><A=20
href=3D"http://www.springerlink.com/content/0428100127358214/fulltext.htm=
l#CR6">6</A></CITE>,=20
<CITE><A=20
href=3D"http://www.springerlink.com/content/0428100127358214/fulltext.htm=
l#CR11">11</A></CITE>].=20
In this context, it is interesting to note that (as demonstrated before =
and=20
exemplified by marker D22S430 in Fig.&nbsp;<A=20
href=3D"http://www.springerlink.com/content/0428100127358214/fulltext.htm=
l#Fig1">1</A>d)=20
the meningioma of our patient has no LOH 22 and the retained <I>INI1</I> =
allele=20
carries no additional inactivating mutation [<CITE><A=20
href=3D"http://www.springerlink.com/content/0428100127358214/fulltext.htm=
l#CR2">2</A></CITE>],=20
indicating that this tumour is not the result of inheritance of the =
mutated=20
<I>INI1</I> gene. In contrast, the meningioma displayed genetic changes =
that are=20
characteristic for radiation-induced meningiomas [<CITE><A=20
href=3D"http://www.springerlink.com/content/0428100127358214/fulltext.htm=
l#CR1">1</A></CITE>,=20
<CITE><A=20
href=3D"http://www.springerlink.com/content/0428100127358214/fulltext.htm=
l#CR7">7</A></CITE>,=20
<CITE><A=20
href=3D"http://www.springerlink.com/content/0428100127358214/fulltext.htm=
l#CR9">9</A></CITE>],=20
such as a complex karyotype, as evidenced by the SNP analysis =
(Table&nbsp;<A=20
href=3D"http://www.springerlink.com/content/0428100127358214/fulltext.htm=
l#Tab1">1</A>),=20
including extensive losses on chromosome arm 1p, but no loss of =
chromosome 22,=20
and no mutation in the <I>NF2</I> gene [<CITE><A=20
href=3D"http://www.springerlink.com/content/0428100127358214/fulltext.htm=
l#CR2">2</A></CITE>].=20
The genetic changes found in the meningioma proved not to be present in =
the=20
AT/RT or myoepithelioma (Fig.&nbsp;<A=20
href=3D"http://www.springerlink.com/content/0428100127358214/fulltext.htm=
l#Fig1">1</A>a=96c,=20
and data not shown), demonstrating that the meningioma is genetically =
not=20
related to these tumours and should not be considered as a late and more =

differentiated recurrence or metastasis of the AT/RT. </P>
<P class=3D"">In conclusion, the clinical and genetic characteristics of =
the=20
meningioma clearly indicate that the tumour developed as a consequence =
of the=20
radiation therapy. Recent evidence suggests that long-term survival of =
patients=20
with a childhood AT/RT can occur especially after aggressive treatment =
including=20
radiotherapy [<CITE><A=20
href=3D"http://www.springerlink.com/content/0428100127358214/fulltext.htm=
l#CR10">10</A></CITE>,=20
<CITE><A=20
href=3D"http://www.springerlink.com/content/0428100127358214/fulltext.htm=
l#CR12">12</A></CITE>].=20
The possible late effects of radiation therapy have been considered as =
serious=20
drawbacks of this treatment. This is the first case in literature in =
which such=20
a possible consequence in the form of a radiation-induced meningioma is =
actually=20
documented in a long-term survivor of an AT/RT. Compared to sporadic=20
meningiomas, radiation-induced meningiomas are more aggressive and have =
a higher=20
chance to recur [<CITE><A=20
href=3D"http://www.springerlink.com/content/0428100127358214/fulltext.htm=
l#CR1">1</A></CITE>].=20
Because of this unfavourable prognosis, continuous medical surveillance =
of our=20
patient is warranted. </P></DIV>
<DIV class=3DAcknowledgments><SPAN=20
class=3DAcknowledgmentsHeading>Acknowledgement&nbsp;&nbsp;</SPAN><SPAN =
class=3D"">We=20
gratefully acknowledge the financial support from the Dutch Cancer =
Society,=20
grants KUN 2003-2975 (PW) and UVA 2001-2561 (TH).</SPAN>
<DIV class=3DFormalPara>
<DIV class=3D""><SPAN style=3D"FONT-STYLE: italic; TEXT-DECORATION: =
none">Open=20
Access</SPAN>&nbsp;&nbsp; This article is distributed under the terms of =
the=20
Creative Commons Attribution Noncommercial License which permits any=20
noncommercial use, distribution, and reproduction in any medium, =
provided the=20
original author(s) and source are credited. </DIV></DIV></DIV>
<P></P>
<HR>

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