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Pediatric Brain Tumors: Signatures from the Intact Proteome

The present investigation aimed to explore the intact proteome of tissues of pediatric brain tumors of different WHO grades and localizations, including medulloblastoma, pilocytic astrocytoma, and glioblastoma, in comparison with the available data on ependymoma, to contribute to the understanding o...

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Autores principales: Rossetti, Diana Valeria, Inserra, Ilaria, Nesticò, Alessia, Vincenzoni, Federica, Iavarone, Federica, Messana, Irene, Castagnola, Massimo, Massimi, Luca, Tamburrini, Gianpiero, Caldarelli, Massimo, Desiderio, Claudia
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8949132/
https://www.ncbi.nlm.nih.gov/pubmed/35328618
http://dx.doi.org/10.3390/ijms23063196
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author Rossetti, Diana Valeria
Inserra, Ilaria
Nesticò, Alessia
Vincenzoni, Federica
Iavarone, Federica
Messana, Irene
Castagnola, Massimo
Massimi, Luca
Tamburrini, Gianpiero
Caldarelli, Massimo
Desiderio, Claudia
author_facet Rossetti, Diana Valeria
Inserra, Ilaria
Nesticò, Alessia
Vincenzoni, Federica
Iavarone, Federica
Messana, Irene
Castagnola, Massimo
Massimi, Luca
Tamburrini, Gianpiero
Caldarelli, Massimo
Desiderio, Claudia
author_sort Rossetti, Diana Valeria
collection PubMed
description The present investigation aimed to explore the intact proteome of tissues of pediatric brain tumors of different WHO grades and localizations, including medulloblastoma, pilocytic astrocytoma, and glioblastoma, in comparison with the available data on ependymoma, to contribute to the understanding of the molecular mechanisms underlying the onset and progression of these pathologies. Tissues have been homogenized in acidic water–acetonitrile solutions containing proteases inhibitors and analyzed by LC–high resolution MS for proteomic characterization and label-free relative quantitation. Tandem MS spectra have been analyzed by either manual inspection or software elaboration, followed by experimental/theoretical MS fragmentation data comparison by bioinformatic tools. Statistically significant differences in protein/peptide levels between the different tumor histotypes have been evaluated by ANOVA test and Tukey’s post-hoc test, considering a p-value > 0.05 as significant. Together with intact protein and peptide chains, in the range of molecular mass of 1.3–22.8 kDa, several naturally occurring fragments from major proteins, peptides, and proteoforms have been also identified, some exhibiting proper biological activities. Protein and peptide sequencing allowed for the identification of different post-translational modifications, with acetylations, oxidations, citrullinations, deamidations, and C-terminal truncations being the most frequently characterized. C-terminal truncations, lacking from two to four amino acid residues, particularly characterizing the β-thymosin peptides and ubiquitin, showed a different modulation in the diverse tumors studied. With respect to the other tumors, medulloblastoma, the most frequent malignant brain tumor of the pediatric age, was characterized by higher levels of thymosin β4 and β10 peptides, the latter and its des-IS form particularly marking this histotype. The distribution pattern of the C-terminal truncated forms was also different in glioblastoma, particularly underlying gender differences, according to the definition of male and female glioblastoma as biologically distinct diseases. Glioblastoma was also distinguished for the peculiar identification of the truncated form of the α-hemoglobin chain, lacking the C-terminal arginine, and exhibiting oxygen-binding and vasoconstrictive properties different from the intact form. The proteomic characterization of the undigested proteome, following the top-down approach, was challenging to originally investigate the post-translational events that differently characterize pediatric brain tumors. This study provides a contribution to elucidate the molecular profiles of the solid tumors most frequently affecting the pediatric age, and which are characterized by different grades of aggressiveness and localization.
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spelling pubmed-89491322022-03-26 Pediatric Brain Tumors: Signatures from the Intact Proteome Rossetti, Diana Valeria Inserra, Ilaria Nesticò, Alessia Vincenzoni, Federica Iavarone, Federica Messana, Irene Castagnola, Massimo Massimi, Luca Tamburrini, Gianpiero Caldarelli, Massimo Desiderio, Claudia Int J Mol Sci Article The present investigation aimed to explore the intact proteome of tissues of pediatric brain tumors of different WHO grades and localizations, including medulloblastoma, pilocytic astrocytoma, and glioblastoma, in comparison with the available data on ependymoma, to contribute to the understanding of the molecular mechanisms underlying the onset and progression of these pathologies. Tissues have been homogenized in acidic water–acetonitrile solutions containing proteases inhibitors and analyzed by LC–high resolution MS for proteomic characterization and label-free relative quantitation. Tandem MS spectra have been analyzed by either manual inspection or software elaboration, followed by experimental/theoretical MS fragmentation data comparison by bioinformatic tools. Statistically significant differences in protein/peptide levels between the different tumor histotypes have been evaluated by ANOVA test and Tukey’s post-hoc test, considering a p-value > 0.05 as significant. Together with intact protein and peptide chains, in the range of molecular mass of 1.3–22.8 kDa, several naturally occurring fragments from major proteins, peptides, and proteoforms have been also identified, some exhibiting proper biological activities. Protein and peptide sequencing allowed for the identification of different post-translational modifications, with acetylations, oxidations, citrullinations, deamidations, and C-terminal truncations being the most frequently characterized. C-terminal truncations, lacking from two to four amino acid residues, particularly characterizing the β-thymosin peptides and ubiquitin, showed a different modulation in the diverse tumors studied. With respect to the other tumors, medulloblastoma, the most frequent malignant brain tumor of the pediatric age, was characterized by higher levels of thymosin β4 and β10 peptides, the latter and its des-IS form particularly marking this histotype. The distribution pattern of the C-terminal truncated forms was also different in glioblastoma, particularly underlying gender differences, according to the definition of male and female glioblastoma as biologically distinct diseases. Glioblastoma was also distinguished for the peculiar identification of the truncated form of the α-hemoglobin chain, lacking the C-terminal arginine, and exhibiting oxygen-binding and vasoconstrictive properties different from the intact form. The proteomic characterization of the undigested proteome, following the top-down approach, was challenging to originally investigate the post-translational events that differently characterize pediatric brain tumors. This study provides a contribution to elucidate the molecular profiles of the solid tumors most frequently affecting the pediatric age, and which are characterized by different grades of aggressiveness and localization. MDPI 2022-03-16 /pmc/articles/PMC8949132/ /pubmed/35328618 http://dx.doi.org/10.3390/ijms23063196 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Rossetti, Diana Valeria
Inserra, Ilaria
Nesticò, Alessia
Vincenzoni, Federica
Iavarone, Federica
Messana, Irene
Castagnola, Massimo
Massimi, Luca
Tamburrini, Gianpiero
Caldarelli, Massimo
Desiderio, Claudia
Pediatric Brain Tumors: Signatures from the Intact Proteome
title Pediatric Brain Tumors: Signatures from the Intact Proteome
title_full Pediatric Brain Tumors: Signatures from the Intact Proteome
title_fullStr Pediatric Brain Tumors: Signatures from the Intact Proteome
title_full_unstemmed Pediatric Brain Tumors: Signatures from the Intact Proteome
title_short Pediatric Brain Tumors: Signatures from the Intact Proteome
title_sort pediatric brain tumors: signatures from the intact proteome
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8949132/
https://www.ncbi.nlm.nih.gov/pubmed/35328618
http://dx.doi.org/10.3390/ijms23063196
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