Proteomic characterization of pediatric craniopharyngioma intracystic fluid by LC‐MS top‐down/bottom‐up integrated approaches

Abstract

The combination of top-down and bottom-up platforms was utilized for the LC-MS proteomic characterization of the intracystic fluid of adamantinomatous craniopharyngioma pediatric brain tumor disease. Proteins and peptides characterization was achieved by high-resolution LC-ESI-LTQ-Orbitrap-MS analysis while low-resolution LC-ESI-IT-MS was applied for the complete screening of the samples and the evaluation of the protein distribution within patients. Top-down analyses were applied to liquid/liquid extracted samples while bottom-up analyses were performed after trypsin digestion of both untreated and pretreated samples. The two proteomic approaches were complementary for the characterization of the proteome of craniopharyngioma intracystic fluid. Proteins and peptides involved in inflammation, mineralization processes and lipid transport were identified, in agreement with the calcium flecks, cholesterol granules and bone residues characteristic of this fluid. Apolipoprotein A-I, A-II, C-I and J, hemoglobin fragments, ubiquitin, α-2-HS-glycoprotein or fetuin A, α-1-antichymotrypsin, vitamin D binding protein, and α-1-acid glycoprotein were characterized. These data could be relevant for the comprehension of the processes involved in the pathogenesis of the disease and the development of the cyst and could contribute to the individuation of therapeutic targets for the reduction of the cyst volume delaying and/or avoiding invasive surgical treatments.

Molecular pathology of adamantinomatous craniopharyngioma: review and opportunities for practice

“…1,3,16,18,19,22,26,32,34,38,40,43,46,50,60 The complex tissue architecture of ACPs, with a range of cell types (e.g., palisading epithelium, stellate reticulum, clusters, reactive glial tissue, and inflammatory cells), has posed a particular challenge. It is important to interpret the molecular data in the context of the appropriate cell types within the tumor and surrounding reactive tissue.…”

“…47,53,54 Expression of a number of inflammatory mediators, such as IL6, IL1A, tumor necrosis factor (TNF), and α-defensins 1-3, has been identified in cystic fluid, generating interest in the use of immunomodulators for the treatment of cysts. 34,38,40 The histopathological similarities between ACP and odontogenesis and frequently cystic odontogenic tumors such as ameloblastoma and calcifying odontogenic cyst has also been recognized, but the molecular biology of these similarities and links remains relatively underexplored. 59 …”

Running Head: Craniopharyngioma and Immune Response

“…Frequently, adjacent brain tissue is infiltrated by leukocytes and cytokine(s) produced by tumor cells may play a pivotal role in tissue reaction [3,7,10]. Two subtypes could be distinguished: an adamantinomatous (Ada) form that is more common in children, and a papillary (Pa) form that is observed almost exclusively in adults [1].…”

“…IL-1alpha, IL-6 and TNF-alpha has been reported significantly higher in the cyst fluid than in cerebrospinal fluid (CSF) [12], Mori et al [13], observed that IL-6 plays an important role in the inflammatory reaction that occurs in the interface between the craniopharyngioma and the brain parenchyma. α-defensins 1-3 and other proteins and peptides have been involved in inflammation, mineralization processes and lipid transport [10]. Apolipoprotein A-I, A-II, C-I and J, haemoglobin fragments, ubiquitin, α-2-HS-glycoprotein or fetuin A, α-1-antichymotrypsin, vitamin D binding protein, and α-1-acid glycoprotein were also have been characterized in the oil machinery fluid [10].…”

“…α-defensins 1-3 and other proteins and peptides have been involved in inflammation, mineralization processes and lipid transport [10]. Apolipoprotein A-I, A-II, C-I and J, haemoglobin fragments, ubiquitin, α-2-HS-glycoprotein or fetuin A, α-1-antichymotrypsin, vitamin D binding protein, and α-1-acid glycoprotein were also have been characterized in the oil machinery fluid [10]. Thymosins β4 and β10 peptides has been also identified in the intracystic fluid AdaCPs [14].…”