TY - JOUR
T1 - Integrative genetic, epigenetic and pathological analysis of paraganglioma reveals complex dysregulation of NOTCH signaling
AU - Cama, Alessandro
AU - Verginelli, Fabio
AU - Lotti, Lavinia Vittoria
AU - Napolitano, Francesco
AU - Morgano, Annalisa
AU - D'Orazio, Andria
AU - Vacca, Michele
AU - Perconti, Silvia
AU - Pepe, Felice
AU - Romani, Federico
AU - Vitullo, Francesca
AU - Di Lella, Filippo
AU - Visone, Rosa
AU - Mannelli, Massimo
AU - Neumann, Hartmut P H
AU - Raiconi, Giancarlo
AU - Paties, Carlo
AU - Moschetta, Antonio
AU - Tagliaferri, Roberto
AU - Veronese, Angelo
AU - Sanna, Mario
AU - Mariani-Costantini, Renato
N1 - Funding Information:
Acknowledgments This study was funded by Associazione Itali-ana Ricerca sul Cancro (AIrC), grant Ig 9168 (2009) to r.M.C.
PY - 2013/10
Y1 - 2013/10
N2 - Head and neck paragangliomas, rare neoplasms of the paraganglia composed of nests of neurosecretory and glial cells embedded in vascular stroma, provide a remarkable example of organoid tumor architecture. To identify genes and pathways commonly deregulated in head and neck paraganglioma, we integrated high-density genome-wide copy number variation (CNV) analysis with microRNA and immunomorphological studies. Gene-centric CNV analysis of 24 cases identified a list of 104 genes most significantly targeted by tumor-associated alterations. The NOTCH signaling pathway was the most significantly enriched term in the list (P = 0.002 after Bonferroni or Benjamini correction). Expression of the relevant NOTCH pathway proteins in sustentacular (glial), chief (neuroendocrine) and endothelial cells was confirmed by immunohistochemistry in 47 head and neck paraganglioma cases. There were no relationships between level and pattern of NOTCH1/JAG2 protein expression and germline mutation status in the SDH genes, implicated in paraganglioma predisposition, or the presence/absence of immunostaining for SDHB, a surrogate marker of SDH mutations. Interestingly, NOTCH upregulation was observed also in cases with no evidence of CNVs at NOTCH signaling genes, suggesting altered epigenetic modulation of this pathway. To address this issue we performed microarray-based microRNA expression analyses. Notably 5 microRNAs (miR-200a,b,c and miR-34b,c), including those most downregulated in the tumors, correlated to NOTCH signaling and directly targeted NOTCH1 in in vitro experiments using SH-SY5Y neuroblastoma cells. Furthermore, lentiviral transduction of miR-200s and miR-34s in patient-derived primary tympano-jugular paraganglioma cell cultures was associated with NOTCH1 downregulation and increased levels of markers of cell toxicity and cell death. Taken together, our results provide an integrated view of common molecular alterations associated with head and neck paraganglioma and reveal an essential role of NOTCH pathway deregulation in this tumor type.
AB - Head and neck paragangliomas, rare neoplasms of the paraganglia composed of nests of neurosecretory and glial cells embedded in vascular stroma, provide a remarkable example of organoid tumor architecture. To identify genes and pathways commonly deregulated in head and neck paraganglioma, we integrated high-density genome-wide copy number variation (CNV) analysis with microRNA and immunomorphological studies. Gene-centric CNV analysis of 24 cases identified a list of 104 genes most significantly targeted by tumor-associated alterations. The NOTCH signaling pathway was the most significantly enriched term in the list (P = 0.002 after Bonferroni or Benjamini correction). Expression of the relevant NOTCH pathway proteins in sustentacular (glial), chief (neuroendocrine) and endothelial cells was confirmed by immunohistochemistry in 47 head and neck paraganglioma cases. There were no relationships between level and pattern of NOTCH1/JAG2 protein expression and germline mutation status in the SDH genes, implicated in paraganglioma predisposition, or the presence/absence of immunostaining for SDHB, a surrogate marker of SDH mutations. Interestingly, NOTCH upregulation was observed also in cases with no evidence of CNVs at NOTCH signaling genes, suggesting altered epigenetic modulation of this pathway. To address this issue we performed microarray-based microRNA expression analyses. Notably 5 microRNAs (miR-200a,b,c and miR-34b,c), including those most downregulated in the tumors, correlated to NOTCH signaling and directly targeted NOTCH1 in in vitro experiments using SH-SY5Y neuroblastoma cells. Furthermore, lentiviral transduction of miR-200s and miR-34s in patient-derived primary tympano-jugular paraganglioma cell cultures was associated with NOTCH1 downregulation and increased levels of markers of cell toxicity and cell death. Taken together, our results provide an integrated view of common molecular alterations associated with head and neck paraganglioma and reveal an essential role of NOTCH pathway deregulation in this tumor type.
KW - CNV
KW - Head and neck
KW - MicroRNA
KW - NOTCH signaling
KW - Paraganglioma
KW - Paraganglioma cell culture
UR - http://www.scopus.com/inward/record.url?scp=84885468590&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84885468590&partnerID=8YFLogxK
U2 - 10.1007/s00401-013-1165-y
DO - 10.1007/s00401-013-1165-y
M3 - Article
C2 - 23955600
AN - SCOPUS:84885468590
SN - 0001-6322
VL - 126
SP - 575
EP - 594
JO - Acta Neuropathologica
JF - Acta Neuropathologica
IS - 4
ER -