Dataset Information

Breaking the mucin barrier: a new affinity chromatography-mass spectrometry approach to unveil potential cell markers and pathways altered in Pseudomyxoma peritonei

ABSTRACT: Pseudomyxoma peritonei (PMP) is a rare, malignant tumour with unknown underlying molecular mechanisms characterised by a progressive accumulation of mucin and secreting tumour cells within the abdomen and pelvis. Cytoreductive surgery (CRS) combined with hyperthermic intraperitoneal chemotherapy (HIPEC) is the only therapeutic option currently available for its treatment and recurrence with a fatal outcome is common despite its use. The lack of information available about the molecular mechanisms underlying this condition is mainly due to the physicochemical characteristics of mucin, whose main purpose is to protect epithelial cells, which makes it inaccessible to a proteomic analysis. In this paper we describe the first protocol available to break this barrier and offer a chance to isolate proteins from soft and hard mucinous tumour tissues. This approach is based on the depletion of glycoproteins, immunoglobins, and albumins by affinity liquid chromatography, which makes the mucin available for label free quantification by mass spectrometry. As in the case of non mucinous tissues, our protocol yields a mix of total proteins and has allowed us to establish the first protein profile in mucinous tumours. In addition, our protein-protein interaction networks and pathway enrichment analyses revealed MUC13 and TFF1 as potential therapeutic targets in this disease.

INSTRUMENT(S): TripleTOF 6600

ORGANISM(S): Homo Sapiens (human) Trypanosoma Cruzi Cruzi

TISSUE(S): Peritoneal Cavity

DISEASE(S): Pseudomyxoma Peritonei

SUBMITTER: Luz Valero

LAB HEAD: Antonio Romero-Ruiz

PROVIDER: PXD037364 | Pride | 2024-06-22

REPOSITORIES: Pride

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Breaking the Mucin Barrier: A New Affinity Chromatography-Mass Spectrometry Approach to Unveil Potential Cell Markers and Pathways Altered in Pseudomyxoma Peritonei.

Romero-Ruiz Antonio A Granados-Rodríguez Melissa M Bura Florina I FI Valenzuela-Molina Francisca F Rufián-Andújar Blanca B Martínez-López Ana A Rodríguez-Ortiz Lidia L Ortega-Salas Rosa R Torres-Martínez María M Moreno-Serrano Ana A Castaño Justo J Michán Carmen C Alhama José J Vázquez-Borrego Mari C MC Arjona-Sánchez Álvaro Á

Biological procedures online 20240515 1

<h4>Background</h4>Pseudomyxoma peritonei (PMP) is a rare peritoneal mucinous carcinomatosis with largely unknown underlying molecular mechanisms. Cytoreductive surgery combined with hyperthermic intraperitoneal chemotherapy is the only therapeutic option; however, despite its use, recurrence with a fatal outcome is common. The lack of molecular characterisation of PMP and other mucinous tumours is mainly due to the physicochemical properties of mucin.<h4>Results</h4>This manuscript describes th ...[more]

PMID: 38750435

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Project description:Carotenoids are naturally occurring pigments in plants responsible for the orange, yellow, and red color of fruits and vegetables. Carrots are one of the primary dietary sources of carotenoids. The biological activities of carotenoids in higher organisms are well documented in most tissues but not the large intestine. The gastrointestinal barrier acts as a line of defense against the systemic invasion of pathogenic bacteria, especially at the colonic level. Proteins involved in tight junction assembly between epithelial cells and mucus secretion from goblet cells are essential for maintaining intestinal barrier homeostasis. A high-fat diet can cause gut impairment by inducing barrier permeability, leading to low-grade chronic inflammation via metabolic endotoxemia. Our hypothesis for this study is that the dietary intake of carotenoid-rich foods can alleviate obesity-associated gut inflammation and strengthen the intestinal barrier function. Male C57BL/6J mice were randomized to one of four experimental diets for 20 weeks (n = 20 animals/group): Low-fat diet (LFD, 10% calories from fat), high-fat diet (HFD, 45% calories from fat), HFD with white carrot powder (HFD + WC), or HFD with orange carrot powder (HFD + OC). Colon tissues were harvested to analyze the biochemical effects of carotenoids in carrots. The distal sections were subjected to isobaric labeling-based quantitative proteomics in which tryptic peptides were labeled with tandem mass tags, followed by fractionation and LC-MS/MS analysis in an Orbitrap Eclipse Tribrid instrument. High-performance liquid chromatography results depicted that the HFD+WC pellets were carotenoid-deficient, and the HFD+OC pellets contained high concentrations of provitamin A carotenoids, specifically α-carotene and β-carotene. As a result of the quantitative proteomics, a total of 4410 differentially expressed proteins were identified. Intestinal barrier-associated proteins were highly upregulated in the HFD+OC group, particularly mucin-2 (MUC-2). Upon closer investigation into mucosal activity, other proteins related to MUC-2 functionality and tight junction management were upregulated by the HFD+OC dietary intervention. Carotenoid-rich foods may prevent high-fat diet-induced intestinal barrier disruption by promoting colonic mucus synthesis and secretion in mammalian organisms.

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	AR_210127-L.1.wiff	Wiff
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