title: Integrative genetic, epigenetic and pathological analysis of paraganglioma reveals complex dysregulation of NOTCH signaling.
abstract: 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.
authors: Unit of General Pathology, Aging Research Center (Ce.S.I.), G. d'Annunzio University Foundation, Via Colle dell'Ara, 66100, Chieti, Italy.
authors: Alessandro
authors: A
authors: Cama
authors: Fabio
authors: F
authors: Verginelli
authors: Lavinia Vittoria
authors: LV
authors: Lotti
authors: Francesco
authors: F
authors: Napolitano
authors: Annalisa
authors: A
authors: Morgano
authors: Andria
authors: A
authors: D'Orazio
authors: Michele
authors: M
authors: Vacca
authors: Silvia
authors: S
authors: Perconti
authors: Felice
authors: F
authors: Pepe
authors: Federico
authors: F
authors: Romani
authors: Francesca
authors: F
authors: Vitullo
authors: Filippo
authors: F
authors: di Lella
authors: Rosa
authors: R
authors: Visone
authors: Massimo
authors: M
authors: Mannelli
authors: Hartmut P H
authors: HP
authors: Neumann
authors: Giancarlo
authors: G
authors: Raiconi
authors: Carlo
authors: C
authors: Paties
authors: Antonio
authors: A
authors: Moschetta
authors: Roberto
authors: R
authors: Tagliaferri
authors: Angelo
authors: A
authors: Veronese
authors: Mario
authors: M
authors: Sanna
authors: Renato
authors: R
authors: Mariani-Costantini
mesh_terms: D015153:Blotting, Western; D020169:Caspases / Q000378:metabolism; D016923:Cell Death / Q000235:genetics; D045744:Cell Line, Tumor; D004252:DNA Mutational Analysis; D044127:Epigenesis, Genetic / Q000502:physiology*; D005455:Fluorescent Antibody Technique; D006258:Head and Neck Neoplasms / Q000235:genetics* / Q000473:pathology*; D006801:Humans; D007150:Immunohistochemistry; D016086:Lentivirus / Q000235:genetics; D046228:Microarray Analysis; D016253:Microscopy, Immunoelectron; D010235:Paraganglioma / Q000235:genetics* / Q000473:pathology*; D010525:Peripheral Nerves / Q000378:metabolism; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D051880:Receptors, Notch / Q000235:genetics* / Q000502:physiology*; D015398:Signal Transduction / Q000235:genetics* / Q000502:physiology*; D013385:Succinate Dehydrogenase / Q000235:genetics; D014162:Transfection
chemical_list: D012333:RNA, Messenger; D051880:Receptors, Notch; D013385:Succinate Dehydrogenase; D020169:Caspases
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