<title>1. INTRODUCTION</title> <p>Malignant peripheral nerve sheath tumours (MPNSTs) are Schwann cell neoplasms that are highly aggressive, frequently lethal, and generally resistant to conventional radiation and chemotherapy [##REF##11894862##1##, ##REF##12951587##2##].</p> <p>Nearly half of these tumours arise in the context of
the inherited predisposition syndrome, neurofibromatosis type 1 (NF1),
suggesting that inactivation of the <italic>NF1</italic> tumour suppressor gene might be causally related to the development of these
cancers [##UREF##0##3##]. NF1 is a dominantly inherited human disease affecting one in 2500 to
3500 individuals [##REF##3145091##4##]. NF1 is characterized by café-au-lait spots (flat pigmented skin
lesions), Lish nodules (abnormality of the iris), skeletal abnormalities,
learning disabilities, neurofibromas, and increased risk of developing
malignant tumours of the central and peripheral nervous system [##UREF##1##5##]. NF1 is associated with mutations of the tumour suppressor gene <italic>NF1</italic>, which encodes for the
Ras-GTPase-activating protein neurofibromin [##REF##2121371##6##–##REF##2134734##8##].</p> <p>Molecular events contributing to peripheral nerve tumour
development are unclear. In the context of NF1, loss of neurofibromin, the NF1
protein product, is believed to be the earliest event, as patients inherit a
mutated <italic>NF1</italic> allele and lose the
second copy in the MPNST cells. Loss of both copies was also observed in benign
neurofibromas. It is likely that tumour suppressor mutations alone are not
sufficient, and that deregulation and/or mutations of oncogenes are necessary
to induce malignant transformation of Schwann cells. The overexpression or
mutation of the tumour suppressor gene <italic>TP53</italic> observed in MPNSTs supports the notion that p53 alterations play a role in
their development [##REF##11453810##9##]. Several studies have demonstrated the central role of epidermal growth
factor receptor (EGFR) in malignant transformation of Schwann cells [##REF##10791998##10##–##REF##15897877##13##]. To our knowledge, only 12 cases of human MPNST have
been studied for EGFR by immunohistochemistry [##REF##10791998##10##, ##REF##15897877##13##]. In the present study, we analyzed the expression of EGFR in the tumours
of 52 patients with MPNST, and compared it with NF1 status and survival.</p>
<title>2. MATERIALS AND METHODS</title> <title>2.1. Patients and samples</title> <p>Patients of the main series (<italic>n</italic> = 52) were all treated in
the Institut Gustave Roussy (IGR, Villejuif, France) between
1985 and 2005. Clinical records were reviewed by one of us (R. Bahleda), with
special attention to initial localization, NF1 status, treatment and survival.
Diagnosis of NF1 was established according to the NIH criteria [##REF##3128965##14##]. Most of the patients had undergone surgery in another centre and were
secondary referred to IGR. Tumours were considered as local stage, when R0
surgery was performed initially, and locally advanced stage for R1 and R2
surgery. Only cases with paraffin embedded MPNST samples were included in the
study. Histological review was realized for all included patients by at least
two pathologists (PT, MJTL, JFE) on hematoxylin-eosin stained slides. Diagnosis
of MPNST was performed according to WHO criteria [##REF##17618441##15##]. Grading of the tumours was not performed, due to limited amounts of
paraffin embedded samples. Immunostaining with S100 protein (rabbit polyclonal,
Dako, Carpenteria, Calif, USA)
and KIT (rabbit polyclonal, Dako) was performed when necessary to confirm
diagnosis.</p> <p>All 52 paraffin embedded samples were subjected to
immunohistochemistry; 8 of which were also analysed by FISH/CISH.</p> <p>Frozen samples from 20 other patients with MPNST were
used for the RNA analysis. Sixteen were from a previously published series [##REF##14723797##16##] and four from Léon Bérard Centre (Lyon, France). Frozen control samples
from 14 patients with benign dermal neurofibromas were also analyzed.</p> <p>All samples were obtained from surgery performed for
diagnostic and/or therapeutic purpose, and were used according to French
ethical regulations.</p> <title>2.2. Immunohistochemistry</title> <p>Immunohistochemistry was performed on four micron
sections from paraffin embedded tumour samples, after antigen retrieval by
heating at 95°C for 20 minutes in 10 mM citrate buffer pH6. For mouse monoclonal
anti-EGFR (31G7, Zymed, South San Francisco, Calif, USA, final dilution 1/10), P53 (DO-7,
Novocastra, Newcastle upon Tyne, UK, final dilution 1/50), and anti-Ki-67 (Mib1 Dako, final dilution
1/50), staining was revealed with LSAB kit (Dako). For anti-Survivin (12C4,
Dako, final dilution 1/100) staining was revealed with CSAII (Dako), according
to manufacturer's instruction.</p> <p>For EGFR staining, tumour cells were considered
negative, when positive signals were detected on nontumour cells (usually
spindle cells and/or small nerves in the periphery of the tumours); otherwise,
staining was considered as not interpretable.</p> <title>2.3. Fluorescent in situ hybridization (FISH)</title> <p>Eight paraffin embedded samples of the main series
were analyzed for EGFR amplification. EGFR specific sequence probe (LSI EGFR)
and control chromosome enumeration probe 7 (CEP7) were used according to the
manufacturers' recommended protocol (Vysis-Abbott Molecular Diagnostics, Baar, Switzerland),
but with some minor modifications. The DNA probes and the sections of tissues
were denatured at 85°C for 5 minutes using a HYBrite instrument. An additional wash in distilled water
was added before counterstaining and mounting with a solution of 4,
6-diamidino-2-phenylindole (DAPI). The results are reported as the ratio of
average EGFR/CEP7 signals per nucleus. Signal ratios of &lt;2 were classified
as nonamplified (NA) and ≥2 as amplified (A). In each section, at least 30
nuclei were counted for signals.</p> <title>2.4. Chromogenic in situ hybridization (CISH)</title> <p>CISH experiments were performed, according to the
protocol given by the supplier (Zymed), along with FISH to have more
information about sample morphology and to have a permanent signal. Results
were interpreted as indicated above for FISH.</p> <title>2.5. Real-time PCR</title> <p>The
theoretical and practical aspects of real-time quantitative RT-PCR using the
ABI Prism 7700 sequence detection system (Applied Biosystems, Foster City, Calif, USA) have been described in detail elsewhere [##REF##14723797##16##].</p> <p>The precise amount of total RNA added to each reaction
mix (based on optical density) and its quality (i.e., lack of extensive
degradation) are both difficult to assess. We therefore also quantified
transcripts of the endogenous RNA control gene <italic>TBP</italic> (Genbank accession NM_003194), which encodes the TATA
box-binding protein. Each sample was normalized on the basis of its <italic>TBP</italic> content. Results, expressed as
N-fold differences in target gene expression relative to the <italic>TBP</italic> gene, and termed
“Ntarget,” were determined as Ntarget = 2Δ<sup>Ctsample</sup>, where the Δ<sup>Ct</sup> value of the sample was
determined by subtracting the average Ct value of the target gene from the
average Ct value of the <italic>TBP</italic> gene.</p> <p>The N<italic>target</italic> values of the samples were subsequently
normalized such that the mean of the dermal neurofibroma N<italic>target</italic> values
was 1.</p> <title>2.6. Statistical analysis</title> <p>Quantitative data were expressed as mean ± standard
deviation; qualitative data as frequency and percent. Comparisons of means were
performed using Student's <italic>t</italic>-test or Mann and Whitney nonparametric test
when necessary. Comparisons of frequencies were performed using the Chi square
test, or Fisher's exact test when necessary.</p> <p>Log-rank tests were used to examine the relationship
between overall survival and the following variables: age, gender, initial localization,
NF1 status, and EGFR expression. Variables with a statistical <italic>P</italic> value &lt;.20 were entered into a Cox model multivariate analysis. <italic>P</italic> value
less than 0.05 was considered significant in multivariate analysis.</p> <p>Statistical analyses were performed with the SAS 8.2
software package (SAS Institute Inc, Cary, NC, USA).</p>
<title>3. RESULTS</title> <p>The mains clinical characteristics of the 52 patients
with MPNST are presented in ##TAB##0##Table 1##. The mean age at time of diagnosis was 23 ± 15 years, and the sex ratio was 30 m/22 f. Tumours were localized in trunk, head
or neck (<italic>n</italic> = 24), or in the limbs (<italic>n</italic> = 28). Half of the patients (<italic>n</italic> = 26) had a NF1,
of whom nine had a familial history of NF1. The age at diagnosis of MPNSTs was
earlier in patients with NF1 (19 ± 9 years) than in non-NF1 patients (27 ± 18
years) (<italic>P</italic> = .04).</p> <p>EGFR expression by tumour cells was detected by
immunohistochemistry in 36 out of the 42 (86%) valuable patients with MPNST;
percentages were higher in the NF1 subgroup (95% versus 75%; <italic>P</italic> = .06; Chi
square test) (see ##TAB##0##Table 1##). Localization of EGFR within tumour cells was either
membranous, cytoplasmic, or both (see ##FIG##0##Figure 1##). In six cases, tumour cells
were negative and 10 other cases were not valuable and were thus excluded from
the analysis.</p> <p>Interestingly, the staining was heterogeneous
throughout the tumour in several cases (see ##FIG##1##Figure 2(a)##). In these cases, EGFR-positive cells were localized in “high-grade” areas, defined as areas
with high-cellular density and high mitotic index. In four of these cases,
samples were available to perform serial sections, and we confirmed that EGFR-positive areas segregate with high-grade features, as proliferative index
detected by Ki-67 expression, and in two cases also with P53-positive areas (see
##FIG##1##Figure 2(b)##). By contrast, staining with survivin, which was positive in all
the cases of MPNST, was diffuse to all tumour areas (see ##FIG##2##Figure 3##).</p> <p>To confirm, by another way, the high frequency of EGFR
overexpression in MPNSTs, we quantified <italic>EGFR</italic> transcripts in an independent series of 20 MPNSTs using real time RT-PCR, and
compared it to 14 benign dermal neurofibromas. The mean of <italic>EGFR</italic> RNA level was higher in MPNSTs than in benign dermal
neurofibromas (1.68 ± 2.5 versus 1 ± 0.4, NS), and four (25%) of MPNST samples
showed marked increases of <italic>EGFR</italic> transcripts
(more than 3 times higher than the mean for benign dermal neurofibromas).</p> <p>To determine whether overexpression of EGFR protein
and RNA might be related to an amplification of <italic>EGFR</italic> locus, we performed CISH and FISH analysis on eight MPNST
samples of the main series (R72, R74, R78, R84, R85, R86, R92, R94, R111,
R116), whose expression was either homogeneous (<italic>n</italic> = 2) or heterogeneous (<italic>n</italic> = 6).
None of the tumour had evidence of <italic>EGFR</italic> amplification. The mean number of spot detected in the nuclei of 40 to 60
tumour cells by sample stained by CISH was 2.42 [range from 2.1 to 3.3]. In only
one case, 5 to 6 spots were detected in some tumour cells, however FISH
revealed a polysomy of chromosome 7 for this tumour (see ##FIG##3##Figure 4##).</p> <p>All the patients underwent surgical resection of the tumour,
except two whose diagnosis was performed at metastatic stage. Two other
patients were lost of view, few days after initial diagnosis, and were thus
excluded for survival analysis. Kaplan-Meyer analysis of overall survival
revealed that the local stage (local or locally advanced) as well as NF1 status had a poor outcome (<italic>P</italic> = .0005 and <italic>P</italic> = .008, resp.), while age
at diagnosis, gender, and EGFR expression had not. Multivariate analysis
revealed that only NF1 status persisted as an independent prognostic factor (<italic>P</italic> = .02), with a hazard ratio at 2.7 [1.2–6.2]. Median survivals of patients with or without NF1
were 18 and 43 months, respectively, and the 5-year survival was 11% and 45%,
respectively.</p>
<title>4. DISCUSSION</title> <p>In this well-defined series of 52 patients with
MPNSTs, we have shown by immunohistochemistry that EGFR was expressed in 86% (71–94%) of cases. In
the independent series of 20 MPNST RNAs, we observed marked <italic>EGFR</italic> RNA overexpression in 4 (25%)
MPNSTs (&gt;3 times the levels in benign dermal neurofibromas). Our results
confirm previous detection of EGFR in 8/12 cases by immunohistochemistry [##REF##10791998##10##, ##REF##15897877##13##], in 6/7 patients by western blotting [##REF##15897877##13##], as well as <italic>EGFR</italic> mRNA in
16/42 cases [##REF##15446585##17##]. In the latter study, NF1 patients were more frequently positive for <italic>EGFR</italic> RNA expression (12/25 versus 4/17
in non NF1 patients). Analysis of human MPNST cell lines also revealed a
stronger and more diffuse expression of EGFR in cells lines derived from NF1 as
compared to non-NF1 patients [##REF##16510576##18##]. EGFR was also more frequently expressed in NF1 patients in our series
(95% versus 75%; <italic>P</italic> = .03). Immunohistochemical EGFR detection data has
been reported in numerous publications with good staining sensitivity and
specificity on paraffin embedded tissue samples.</p> <p>Overexpression of both protein and RNA suggests
pretranslational regulation of EGFR in MPNSTs. Amplification of gene locus is a
common mechanism of regulation of EGFR in other tumours such as head and neck
squamous cell carcinomas [##REF##16943533##19##], non-small-cell lung carcinomas [##REF##16131043##20##], and colorectal carcinomas [##REF##15863375##21##]. In MPNSTs, <italic>EGFR</italic> amplification has previously been reported in 5 out of 17 patients [##REF##12152785##22##]. In this study, a “low-level” amplification was described with
scattered cells containing 6–12 spots,
accompanied by polysomy 7 in three cases. Another group failed to detect <italic>EGFR</italic> amplification in four cases, although 1-2 extra copies
were seen in one of these cases [##REF##15509679##23##]. Here, we confirm these latter results in eight patients. Thus, EGFR overexpression
in the majority of MPNSTs is not due to amplification of the <italic>EGFR</italic> locus at 7p12. Normal Schwann
cells do not express EGFR, while NF1 mutation leads to EGFR overexpression in
these cells [##REF##15652750##12##]. NF1 loss of function may thus enhance
transcription of EGFR.</p> <p>Mice with heterozygous deletions of <italic>NF1</italic> (<italic>Nf1<sup>+/−</sup></italic>)
do not have an increased incidence of nerve tumours; however when these mice
also carry a heterozygous mutations of <italic>TP53</italic> (<italic>Nf1<sup>+/−</sup><italic>p</italic>53<sup>+/−</sup></italic>) they develop
sarcomas and brain tumours [##REF##10591652##24##, ##REF##10591653##25##]. EGFR is frequently expressed in Schwann cell lines derived from these
(<italic>Nf1<sup>+/−</sup><italic>p</italic>53<sup>+/−</sup></italic>) mice. Cell growth in
these lines is greatly stimulated by EGF and blocked by EGFR antagonists [##REF##12154062##11##]. Decreased EGFR signaling in <italic>Nf1<sup>+/−</sup><italic>p</italic>53<sup>+/−</sup></italic> mice reduced their mortality [##REF##15652750##12##]. In the present series, we showed that high expression of EGFR was
present in the high-grade areas of the tumours, appearing to colocalize with Ki-67 in all cases and with p53
in half of the cases. In several cases, a strong EGFR expression of in highly
cellular regions, contrasting with negativity in other regions, has already been
described in one NF1 patient [##REF##10791998##10##]. Thus, as for animal and in vitro models, our data suggest that EGFR overexpression
is associated with malignant transformation of Schwann cells.</p> <p>However, NF1 status was the only prognostic factor in
multivariate analysis, with median survivals of 18 and 43 months for patients
with or without NF1. EGFR expression, although higher in NF1 patients, did not
appear as a prognostic factor for MPNST, nor did local stage, age at diagnosis,
or gender.</p> <p>Overexpression of <italic>survivin</italic> mRNA in MPNSTs has been observed independently by three
groups [##REF##14723797##16##, ##REF##15446585##17##, ##REF##16443732##26##]. Supervised analysis of gene expression profiling of
MPNSTs revealed that <italic>EGFR</italic>-positive
and -negative tumours had a specific gene expression signatures [##REF##15446585##17##]. Interestingly, these authors showed that <italic>EGFR</italic>-positive tumours had a higher expression of <italic>Ki-67</italic> and <italic>survivin</italic> transcripts. We confirmed herein by immunohistochemistry,
that bring supplementary data about cellular localization of the expression,
that survivin was expressed by malignant Schwann cells. But, contrasting with
EGFR, survivin expression was not restricted to high-grade areas of the
tumours.</p> <p>The prognosis of MPNSTs is poor, with only 23% of
living individuals 10 years after diagnosis [##REF##3082508##27##]. Post-surgical irradiation, as no effect on overall survival and no
effective chemotherapeutic regimens, is available [##REF##11894862##1##]. In our series, the mean age of diagnosis was 23 ± 15 years and the
median survival was 30 months. Thus, there is considerable interest in
establishing the mechanisms responsible for MPNST tumourigenesis and using this
information to develop new, more effective, therapies. Targeted therapies using
monoclonal antibodies against EGFR are highly effective in several human cancer
[##REF##15821783##28##]. So far, most of the patients treated with a monoclonal antibody
anti-EGFR Cetuximab (Erbitux, Lyon, France) suffer from colorectal [##REF##16010411##29##] or lung [##REF##16857823##30##] cancers. Recently, Cetuximab was successfully used to prevent the
development of neurofibromas in a mouse model of NF1 [##REF##16651634##31##]. Several groups showed the implication of <italic>EGFR</italic> expression in malignant transformation of Schwann cells in
cell lines and/or mouse models. Our results obtained in a large series of human
MPNSTs confirmed these data. Tumours with tyrosine kinase receptor
overexpression have been successfully treated with targeted therapies, as in
gastrointestinal stromal tumours, which express KIT and may be treated with
Imatinib [##REF##15365079##32##]. Lung adenocarcinomas, in which the expression of EGFR has no
prognostic value [##REF##7952390##33##], may also be treated with gefitinib or erlotinib. Thus, the
overexpression of EGFR in 95% of NF1 patients with MPNST and the very poor
prognosis of these young patients shown in the present study suggest that new
therapies targeting EGFR might be interesting for these patients.</p>
<p>Recommended by Beatrice Seddon</p> <p>Malignant peripheral nerve sheath tumours (MPNSTs) are highly malignant and resistant. Transformation might implicate up regulation of epidermal growth factor receptor (EGFR). Fifty-two MPNST samples were studied for EGFR, Ki-67, p53, and survivin expression by immunohistochemistry and for <italic>EGFR</italic> amplification by in situ hybridization. Results were correlated with clinical data. <italic>EGFR</italic> RNA was also quantified by RT-PCR in 20 other MPNSTs and 14 dermal neurofibromas. Half of the patients had a neurofibromatosis type 1 (NF1). EGFR expression, detected in 86% of MPNSTs, was more frequent in NF1 specimens and closely associated with high-grade and p53-positive areas. MPNSTs expressed more <italic>EGFR</italic> transcripts than neurofibromas. No amplification of <italic>EGFR</italic> locus was observed. NF1 status was the only prognostic factor in multivariate analysis, with median survivals of 18 and 43 months for patients with or without NF1. Finally, EGFR might become a new target for MPNSTs treatment, especially in NF1-associated MPNSTs.</p>
<title>ACKNOWLEDGMENTS</title> <p>The authors would like to thank all the French,
Italian, and Spanish pathologists, who provided paraffin blocks of the
patients: Dr Andre, Baviera, Bendjaballah, Bertin, Blache, Boddaert, Bognel,
Brucher, Cordonnier, Devillebichot, Dulmet, Ferlicot, Gruchy,
Guillermand-Gerard, Hénin, Lacroix-Ciaudo, Lame, Leger-Ravet, Maiorano, Massart
Moreau, Mathieu, Mikol, Molimard, Nistal Martin de Serrano, Payan, Petit,
Ranchère-Vince, Schmid, Slama, Tourneur, Voisin-Rigaud, Weiss and Yaziji. The authors also want to thank
Mariama Bakhari for excellent technical assistance. S. Tabone-Eglinger is a fellow of ANRT, with financial
support of Centre Léon Bérard Cancer and Novartis SA. This work was supported by the Saint-Quentin-en-Yveline University (“Bonus Qualité
Recherche”) and Association pour la Recherche en Pathologie.</p>
<fig id="fig1" position="float"><label>Figure 1</label><caption><p>EGFR expression in MPNSTs. In both cases, 100% of tumour cells strongly expressed EGFR (brown). 
However, it was detected either (a) within the cytoplasm or (b) on plasma membrane. Cell nuclei were stained in blue by hematoxylin. Scale bar represent 20 <italic>μ</italic>m.</p></caption></fig> <fig id="fig2" position="float"><label>Figure 2</label><caption><p>Heterogeneous expression of EGFR (a) and colocalization with high-grade Ki-67 and p53-positive (b) areas. Scale bar represents 15 <italic>μ</italic>m.</p></caption></fig> <fig id="fig3" position="float"><label>Figure 3</label><caption><p>Homogenous expression of survivin in MPNSTs. Scale bar represents 20 <italic>μ</italic>m.</p></caption></fig> <fig id="fig4" position="float"><label>Figure 4</label><caption><p>CISH and FISH analysis on the R84 MPNST sample. (a) The mean number of spot detected in the nuclei of 40 to 60 tumour cells in this sample stained by CISH was 3.3. (b) FISH confirmed multiple spot of <italic>EGFR</italic> (red) but revealed a polysomy of chromosome 7 (green) for this tumour. Scale bar represents 5 <italic>μ</italic>m.</p></caption></fig>
<table-wrap id="tab1" position="float"><label>Table 1</label><caption><p>
<italic>Characteristics of the 52 patients with MPNST</italic>. NF1:
Neurofibromatosis type 1; Spo: sporadic form of NF1 (no familial history), Fam:
familial form of NF1; L: local stage; LA: locally advanced stage; Met: metastatic stage. A: alive. D: dead; +: positive staining; −: negative staining; n.e.: not evaluable staining.</p></caption><table frame="hsides" rules="groups"><thead><tr><th align="left" rowspan="1" colspan="1">Patient <italic>n</italic>°</th><th align="center" rowspan="1" colspan="1">NF1</th><th align="center" rowspan="1" colspan="1">Age/gender</th><th align="center" rowspan="1" colspan="1">Stage init</th><th align="center" rowspan="1" colspan="1">Localization</th><th align="center" rowspan="1" colspan="1">EGFR</th></tr></thead><tbody><tr><td align="left" rowspan="1" colspan="1">R72</td><td align="center" rowspan="1" colspan="1">no</td><td align="center" rowspan="1" colspan="1">37/m</td><td align="center" rowspan="1" colspan="1">LA</td><td align="center" rowspan="1" colspan="1">Neck</td><td align="center" rowspan="1" colspan="1">+</td></tr><tr><td align="left" rowspan="1" colspan="1">R69</td><td align="center" rowspan="1" colspan="1">no</td><td align="center" rowspan="1" colspan="1">12/m</td><td align="center" rowspan="1" colspan="1">LA</td><td align="center" rowspan="1" colspan="1">Thigh</td><td align="center" rowspan="1" colspan="1">+</td></tr><tr><td align="left" rowspan="1" colspan="1">R92</td><td align="center" rowspan="1" colspan="1">no</td><td align="center" rowspan="1" colspan="1">11/w</td><td align="center" rowspan="1" colspan="1">LA</td><td align="center" rowspan="1" colspan="1">Brachial plexus</td><td align="center" rowspan="1" colspan="1">+</td></tr><tr><td align="left" rowspan="1" colspan="1">R78</td><td align="center" rowspan="1" colspan="1">no</td><td align="center" rowspan="1" colspan="1">28/w</td><td align="center" rowspan="1" colspan="1">L</td><td align="center" rowspan="1" colspan="1">Left arm</td><td align="center" rowspan="1" colspan="1">+</td></tr><tr><td align="left" rowspan="1" colspan="1">R94</td><td align="center" rowspan="1" colspan="1">no</td><td align="center" rowspan="1" colspan="1">52/m</td><td align="center" rowspan="1" colspan="1">L</td><td align="center" rowspan="1" colspan="1">Ethmoid</td><td align="center" rowspan="1" colspan="1">+</td></tr><tr><td align="left" rowspan="1" colspan="1">R86</td><td align="center" rowspan="1" colspan="1">no</td><td align="center" rowspan="1" colspan="1">33/f</td><td align="center" rowspan="1" colspan="1">L</td><td align="center" rowspan="1" colspan="1">Arm</td><td align="center" rowspan="1" colspan="1">+</td></tr><tr><td align="left" rowspan="1" colspan="1">R87</td><td align="center" rowspan="1" colspan="1">no</td><td align="center" rowspan="1" colspan="1">3/f</td><td align="center" rowspan="1" colspan="1">L</td><td align="center" rowspan="1" colspan="1">Tibial nerve</td><td align="center" rowspan="1" colspan="1">+</td></tr><tr><td align="left" rowspan="1" colspan="1">R77</td><td align="center" rowspan="1" colspan="1">no</td><td align="center" rowspan="1" colspan="1">71/f</td><td align="center" rowspan="1" colspan="1">L</td><td align="center" rowspan="1" colspan="1">Median nerve</td><td align="center" rowspan="1" colspan="1">+</td></tr><tr><td align="left" rowspan="1" colspan="1">R84</td><td align="center" rowspan="1" colspan="1">no</td><td align="center" rowspan="1" colspan="1">19/m</td><td align="center" rowspan="1" colspan="1">LA</td><td align="center" rowspan="1" colspan="1">Pelvis</td><td align="center" rowspan="1" colspan="1">+</td></tr><tr><td align="left" rowspan="1" colspan="1">R90</td><td align="center" rowspan="1" colspan="1">no</td><td align="center" rowspan="1" colspan="1">47/m</td><td align="center" rowspan="1" colspan="1">L</td><td align="center" rowspan="1" colspan="1">Right thigh</td><td align="center" rowspan="1" colspan="1">+</td></tr><tr><td align="left" rowspan="1" colspan="1">R74</td><td align="center" rowspan="1" colspan="1">no</td><td align="center" rowspan="1" colspan="1">16/f</td><td align="center" rowspan="1" colspan="1">L</td><td align="center" rowspan="1" colspan="1">Foot</td><td align="center" rowspan="1" colspan="1">+</td></tr><tr><td align="left" rowspan="1" colspan="1">R85</td><td align="center" rowspan="1" colspan="1">no</td><td align="center" rowspan="1" colspan="1">48/f</td><td align="center" rowspan="1" colspan="1">L</td><td align="center" rowspan="1" colspan="1">Forearm</td><td align="center" rowspan="1" colspan="1">+</td></tr><tr><td align="left" rowspan="1" colspan="1">R93</td><td align="center" rowspan="1" colspan="1">no</td><td align="center" rowspan="1" colspan="1">7/f</td><td align="center" rowspan="1" colspan="1">L</td><td align="center" rowspan="1" colspan="1">Mandible</td><td align="center" rowspan="1" colspan="1">+</td></tr><tr><td align="left" rowspan="1" colspan="1">R79</td><td align="center" rowspan="1" colspan="1">no</td><td align="center" rowspan="1" colspan="1">30/m</td><td align="center" rowspan="1" colspan="1">L</td><td align="center" rowspan="1" colspan="1">Wrist</td><td align="center" rowspan="1" colspan="1">+</td></tr><tr><td align="left" rowspan="1" colspan="1">R89</td><td align="center" rowspan="1" colspan="1">no</td><td align="center" rowspan="1" colspan="1">18/f</td><td align="center" rowspan="1" colspan="1">LA</td><td align="center" rowspan="1" colspan="1">Infratemporal fossa</td><td align="center" rowspan="1" colspan="1">+</td></tr><tr><td align="left" rowspan="1" colspan="1">R88</td><td align="center" rowspan="1" colspan="1">no</td><td align="center" rowspan="1" colspan="1">26/m</td><td align="center" rowspan="1" colspan="1">L</td><td align="center" rowspan="1" colspan="1">Left calf</td><td align="center" rowspan="1" colspan="1">0</td></tr><tr><td align="left" rowspan="1" colspan="1">R75</td><td align="center" rowspan="1" colspan="1">no</td><td align="center" rowspan="1" colspan="1">50/f</td><td align="center" rowspan="1" colspan="1">LA</td><td align="center" rowspan="1" colspan="1">Retroperitoneum</td><td align="center" rowspan="1" colspan="1">0</td></tr><tr><td align="left" rowspan="1" colspan="1">R76</td><td align="center" rowspan="1" colspan="1">no</td><td align="center" rowspan="1" colspan="1">23/m</td><td align="center" rowspan="1" colspan="1">L</td><td align="center" rowspan="1" colspan="1">Frontal region</td><td align="center" rowspan="1" colspan="1">0</td></tr><tr><td align="left" rowspan="1" colspan="1">B1027</td><td align="center" rowspan="1" colspan="1">no</td><td align="center" rowspan="1" colspan="1">15/m</td><td align="center" rowspan="1" colspan="1">LA</td><td align="center" rowspan="1" colspan="1">Retroperitoneum, pelvis</td><td align="center" rowspan="1" colspan="1">0</td></tr><tr><td align="left" rowspan="1" colspan="1">R73</td><td align="center" rowspan="1" colspan="1">no</td><td align="center" rowspan="1" colspan="1">7/m</td><td align="center" rowspan="1" colspan="1">LA</td><td align="center" rowspan="1" colspan="1">Neck</td><td align="center" rowspan="1" colspan="1">0</td></tr><tr><td align="left" rowspan="1" colspan="1">B1499</td><td align="center" rowspan="1" colspan="1">no</td><td align="center" rowspan="1" colspan="1">60/m</td><td align="center" rowspan="1" colspan="1">LA</td><td align="center" rowspan="1" colspan="1">Mediastinum</td><td align="center" rowspan="1" colspan="1">n.e.</td></tr><tr><td align="left" rowspan="1" colspan="1">R80</td><td align="center" rowspan="1" colspan="1">no</td><td align="center" rowspan="1" colspan="1">14/f</td><td align="center" rowspan="1" colspan="1">L</td><td align="center" rowspan="1" colspan="1">Left orbit</td><td align="center" rowspan="1" colspan="1">n.e.</td></tr><tr><td align="left" rowspan="1" colspan="1">B1044</td><td align="center" rowspan="1" colspan="1">no</td><td align="center" rowspan="1" colspan="1">7/m</td><td align="center" rowspan="1" colspan="1">L</td><td align="center" rowspan="1" colspan="1">Calf</td><td align="center" rowspan="1" colspan="1">n.e.</td></tr><tr><td align="left" rowspan="1" colspan="1">R95</td><td align="center" rowspan="1" colspan="1">no</td><td align="center" rowspan="1" colspan="1">20/m</td><td align="center" rowspan="1" colspan="1">Met</td><td align="center" rowspan="1" colspan="1">Hand</td><td align="center" rowspan="1" colspan="1">n.e.</td></tr><tr><td align="left" rowspan="1" colspan="1">R81</td><td align="center" rowspan="1" colspan="1">no</td><td align="center" rowspan="1" colspan="1">46/m</td><td align="center" rowspan="1" colspan="1">L</td><td align="center" rowspan="1" colspan="1">Forearm</td><td align="center" rowspan="1" colspan="1">n.e.</td></tr><tr><td align="left" rowspan="1" colspan="1">B1463</td><td align="center" rowspan="1" colspan="1">no</td><td align="center" rowspan="1" colspan="1">19/m</td><td align="center" rowspan="1" colspan="1">LA</td><td align="center" rowspan="1" colspan="1">Armpit</td><td align="center" rowspan="1" colspan="1">n.e.</td></tr><tr><td align="left" rowspan="1" colspan="1">R123</td><td align="center" rowspan="1" colspan="1">Fam</td><td align="center" rowspan="1" colspan="1">15/m</td><td align="center" rowspan="1" colspan="1">L</td><td align="center" rowspan="1" colspan="1">Median nerve</td><td align="center" rowspan="1" colspan="1">+</td></tr><tr><td align="left" rowspan="1" colspan="1">R118 B</td><td align="center" rowspan="1" colspan="1">Fam</td><td align="center" rowspan="1" colspan="1">11/f</td><td align="center" rowspan="1" colspan="1">LA</td><td align="center" rowspan="1" colspan="1">Brachial plexus</td><td align="center" rowspan="1" colspan="1">+</td></tr><tr><td align="left" rowspan="1" colspan="1">B1357</td><td align="center" rowspan="1" colspan="1">Spo</td><td align="center" rowspan="1" colspan="1">16/m</td><td align="center" rowspan="1" colspan="1">LA</td><td align="center" rowspan="1" colspan="1">Sciatic nerve</td><td align="center" rowspan="1" colspan="1">+</td></tr><tr><td align="left" rowspan="1" colspan="1">R107</td><td align="center" rowspan="1" colspan="1">Spo</td><td align="center" rowspan="1" colspan="1">33/m</td><td align="center" rowspan="1" colspan="1">L</td><td align="center" rowspan="1" colspan="1">Neck</td><td align="center" rowspan="1" colspan="1">+</td></tr><tr><td align="left" rowspan="1" colspan="1">R111</td><td align="center" rowspan="1" colspan="1">Spo</td><td align="center" rowspan="1" colspan="1">13/f</td><td align="center" rowspan="1" colspan="1">LA</td><td align="center" rowspan="1" colspan="1">Thigh</td><td align="center" rowspan="1" colspan="1">+</td></tr><tr><td align="left" rowspan="1" colspan="1">B1064</td><td align="center" rowspan="1" colspan="1">Spo</td><td align="center" rowspan="1" colspan="1">44/f</td><td align="center" rowspan="1" colspan="1">L</td><td align="center" rowspan="1" colspan="1">Thigh</td><td align="center" rowspan="1" colspan="1">+</td></tr><tr><td align="left" rowspan="1" colspan="1">R116</td><td align="center" rowspan="1" colspan="1">Spo</td><td align="center" rowspan="1" colspan="1">10/m</td><td align="center" rowspan="1" colspan="1">LA</td><td align="center" rowspan="1" colspan="1">Arm</td><td align="center" rowspan="1" colspan="1">+</td></tr><tr><td align="left" rowspan="1" colspan="1">R98</td><td align="center" rowspan="1" colspan="1">Spo</td><td align="center" rowspan="1" colspan="1">23/f</td><td align="center" rowspan="1" colspan="1">L</td><td align="center" rowspan="1" colspan="1">Calf</td><td align="center" rowspan="1" colspan="1">+</td></tr><tr><td align="left" rowspan="1" colspan="1">B2387</td><td align="center" rowspan="1" colspan="1">Spo</td><td align="center" rowspan="1" colspan="1">29/m</td><td align="center" rowspan="1" colspan="1">LA</td><td align="center" rowspan="1" colspan="1">Supraclavicular region</td><td align="center" rowspan="1" colspan="1">+</td></tr><tr><td align="left" rowspan="1" colspan="1">R122</td><td align="center" rowspan="1" colspan="1">Fam</td><td align="center" rowspan="1" colspan="1">19/f</td><td align="center" rowspan="1" colspan="1">LA</td><td align="center" rowspan="1" colspan="1">Groin</td><td align="center" rowspan="1" colspan="1">+</td></tr><tr><td align="left" rowspan="1" colspan="1">R104</td><td align="center" rowspan="1" colspan="1">Spo</td><td align="center" rowspan="1" colspan="1">25/m</td><td align="center" rowspan="1" colspan="1">L</td><td align="center" rowspan="1" colspan="1">Left calf</td><td align="center" rowspan="1" colspan="1">+</td></tr><tr><td align="left" rowspan="1" colspan="1">R97</td><td align="center" rowspan="1" colspan="1">Spo</td><td align="center" rowspan="1" colspan="1">17/m</td><td align="center" rowspan="1" colspan="1">LA</td><td align="center" rowspan="1" colspan="1">Left iliac</td><td align="center" rowspan="1" colspan="1">+</td></tr><tr><td align="left" rowspan="1" colspan="1">B1400</td><td align="center" rowspan="1" colspan="1">Fam</td><td align="center" rowspan="1" colspan="1">20/m</td><td align="center" rowspan="1" colspan="1">LA</td><td align="center" rowspan="1" colspan="1">Chest wall</td><td align="center" rowspan="1" colspan="1">+</td></tr><tr><td align="left" rowspan="1" colspan="1">R105</td><td align="center" rowspan="1" colspan="1">Spo</td><td align="center" rowspan="1" colspan="1">13/m</td><td align="center" rowspan="1" colspan="1">LA</td><td align="center" rowspan="1" colspan="1">Abdominal wall</td><td align="center" rowspan="1" colspan="1">+</td></tr><tr><td align="left" rowspan="1" colspan="1">R96</td><td align="center" rowspan="1" colspan="1">Spo</td><td align="center" rowspan="1" colspan="1">23/m</td><td align="center" rowspan="1" colspan="1">LA</td><td align="center" rowspan="1" colspan="1">Sciatic nerve buttock</td><td align="center" rowspan="1" colspan="1">+</td></tr><tr><td align="left" rowspan="1" colspan="1">R102</td><td align="center" rowspan="1" colspan="1">Spo</td><td align="center" rowspan="1" colspan="1">23/m</td><td align="center" rowspan="1" colspan="1">Méta</td><td align="center" rowspan="1" colspan="1">Retroperitoneum</td><td align="center" rowspan="1" colspan="1">+</td></tr><tr><td align="left" rowspan="1" colspan="1">R101</td><td align="center" rowspan="1" colspan="1">Spo</td><td align="center" rowspan="1" colspan="1">26/m</td><td align="center" rowspan="1" colspan="1">LA</td><td align="center" rowspan="1" colspan="1">Retroperitoneum</td><td align="center" rowspan="1" colspan="1">+</td></tr><tr><td align="left" rowspan="1" colspan="1">B1399</td><td align="center" rowspan="1" colspan="1">Spo</td><td align="center" rowspan="1" colspan="1">5/m</td><td align="center" rowspan="1" colspan="1">LA</td><td align="center" rowspan="1" colspan="1">Urinary bladder</td><td align="center" rowspan="1" colspan="1">+</td></tr><tr><td align="left" rowspan="1" colspan="1">R114</td><td align="center" rowspan="1" colspan="1">Spo</td><td align="center" rowspan="1" colspan="1">7/m</td><td align="center" rowspan="1" colspan="1">LA</td><td align="center" rowspan="1" colspan="1">Thigh</td><td align="center" rowspan="1" colspan="1">+</td></tr><tr><td align="left" rowspan="1" colspan="1">B1148</td><td align="center" rowspan="1" colspan="1">Fam</td><td align="center" rowspan="1" colspan="1">17/f</td><td align="center" rowspan="1" colspan="1">L</td><td align="center" rowspan="1" colspan="1">Thigh</td><td align="center" rowspan="1" colspan="1">+</td></tr><tr><td align="left" rowspan="1" colspan="1">bloc t</td><td align="center" rowspan="1" colspan="1">Spo</td><td align="center" rowspan="1" colspan="1">32/f</td><td align="center" rowspan="1" colspan="1">L</td><td align="center" rowspan="1" colspan="1">Thigh</td><td align="center" rowspan="1" colspan="1">+</td></tr><tr><td align="left" rowspan="1" colspan="1">B1793</td><td align="center" rowspan="1" colspan="1">Fam</td><td align="center" rowspan="1" colspan="1">32/f</td><td align="center" rowspan="1" colspan="1">L</td><td align="center" rowspan="1" colspan="1">Thigh</td><td align="center" rowspan="1" colspan="1">0</td></tr><tr><td align="left" rowspan="1" colspan="1">R127</td><td align="center" rowspan="1" colspan="1">Fam</td><td align="center" rowspan="1" colspan="1">11/f</td><td align="center" rowspan="1" colspan="1">LA</td><td align="center" rowspan="1" colspan="1">Left thigh (sciatic)</td><td align="center" rowspan="1" colspan="1">n.e.</td></tr><tr><td align="left" rowspan="1" colspan="1">R119</td><td align="center" rowspan="1" colspan="1">Fam</td><td align="center" rowspan="1" colspan="1">10/m</td><td align="center" rowspan="1" colspan="1">LA</td><td align="center" rowspan="1" colspan="1">Retroperitoneum</td><td align="center" rowspan="1" colspan="1">n.e.</td></tr><tr><td align="left" rowspan="1" colspan="1">R125</td><td align="center" rowspan="1" colspan="1">Fam</td><td align="center" rowspan="1" colspan="1">11/f</td><td align="center" rowspan="1" colspan="1">LA</td><td align="center" rowspan="1" colspan="1">Upper</td><td align="center" rowspan="1" colspan="1">n.e.</td></tr><tr><td align="left" rowspan="1" colspan="1">B1599</td><td align="center" rowspan="1" colspan="1">Spo</td><td align="center" rowspan="1" colspan="1">20/f</td><td align="center" rowspan="1" colspan="1">LA</td><td align="center" rowspan="1" colspan="1">Pelvis</td><td align="center" rowspan="1" colspan="1">n.e.</td></tr></tbody></table></table-wrap>
<graphic xlink:href="SRCM2008-849156.001"/> <graphic xlink:href="SRCM2008-849156.002"/> <graphic xlink:href="SRCM2008-849156.003"/> <graphic xlink:href="SRCM2008-849156.004"/>
[{"label": ["3"], "surname": ["Friedman"], "given-names": ["JM"], "article-title": ["Epidemiology of neurofibromatosis type 1"], "italic": ["American Journal of Medical Genetics Part C"], "year": ["1999"], "volume": ["89"], "issue": ["1"], "fpage": ["1"], "lpage": ["6"]}, {"label": ["5"], "surname": ["Riccardi"], "given-names": ["VM"], "italic": ["Neurofibromatosis: Phenotype, Natural History, and Pathogenesis"], "year": ["1992"], "publisher-loc": ["Baltimore, Md, USA"], "publisher-name": ["John Hopkins University Press"]}]
Sarcoma. 2008 Aug 28; 2008:849156