Head and Neck Paragangliomas in Von Hippel-Lindau Disease and ...
Head and Neck Paragangliomas in Von Hippel-Lindau Disease and ...
Head and Neck Paragangliomas in Von Hippel-Lindau Disease and ...
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MATERIAL AND METHODS<br />
Subjects<br />
The <strong>in</strong>ternational Non-SDHx-HNP-Consortium was constituted <strong>in</strong> 2008 <strong>in</strong> order to identify <strong>and</strong> to<br />
cl<strong>in</strong>ically <strong>and</strong> genetically characterize patients with HNP with heritable syndromes <strong>in</strong> the absence of<br />
germl<strong>in</strong>e SDHx mutations. We paid particular attention to additional component tumors suggestive of<br />
VHL, MEN 2 or NF 1. We adopted <strong>and</strong> updated the registrants from the European Australian<br />
American HNP Registry <strong>in</strong>clud<strong>in</strong>g symptomatic <strong>and</strong> asymptomatic HNPs as of March 1, 2009 which<br />
<strong>in</strong>cluded a total of 513 subjects. In particular, centers dedicated to cl<strong>in</strong>ical care <strong>and</strong> molecular genetic<br />
diagnosis of HNPs <strong>in</strong> 26 countries <strong>in</strong>clud<strong>in</strong>g Germany, Spa<strong>in</strong>, The Netherl<strong>and</strong>s, Italy, Hungary,<br />
Ukra<strong>in</strong>e, Serbia, Greece, Pol<strong>and</strong>, Sweden, F<strong>in</strong>l<strong>and</strong>, Denmark, Belgium, France, Switzerl<strong>and</strong>, Austria,<br />
Turkey, Iran, Ch<strong>in</strong>a, Japan, the USA, Brazil, Chile, Argent<strong>in</strong>a, Australia <strong>and</strong> New Zeal<strong>and</strong> were<br />
recontacted. Separately to the European Australian American HNP Registry, the Netherl<strong>and</strong>s <strong>and</strong><br />
France have national HNP registries with 147 <strong>and</strong> 149 registrants <strong>in</strong> whom SDHx mutations have been<br />
excluded, respectively. This aimed to identify unrecognized patients with HNP for case registration<br />
<strong>and</strong> for updat<strong>in</strong>g the HNP registry for prevalence estimations.<br />
For prevalence estimations <strong>in</strong> VHL, we used the national population-based registries for Germany,<br />
France, Spa<strong>in</strong>, <strong>and</strong> the Netherl<strong>and</strong>s.<br />
Genetic Analysis<br />
All registrants provided 10 ml EDTA-anticoagulated blood for molecular genetic analyses. The<br />
analyses comprised test<strong>in</strong>g of the VHL (RefSeq:NM_000551.2) <strong>and</strong> RET (RefSeq:NM_020975.4)<br />
genes for germl<strong>in</strong>e mutations. DNA extraction, primers, PCR conditions <strong>and</strong> sequenc<strong>in</strong>g methods<br />
followed published protocols of <strong>in</strong>ternational st<strong>and</strong>ards (1, 2, 10, 11). Of note, analyses also <strong>in</strong>cluded<br />
screen<strong>in</strong>g for large deletions or rearrangements of the VHL gene, s<strong>in</strong>ce such mutations have been<br />
described <strong>in</strong> VHL patients with pheochromocytoma (13, 14). Overall, 809 HNP patients<br />
(513+147+149) of the European Australian American HNP Registry, the French HNP Registry <strong>and</strong><br />
the Rotterdam HNP Registry comb<strong>in</strong>ed were subjected to the above described analyses.<br />
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