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Proteomics analysis of High-grade glioma tissues showing differential fluorescence following 5-Aminolevulinic acid guided surgery

ABSTRACT: Targeted treatment of high-grade gliomas (HGGs) is challenging due to intra- and inter-tumoral heterogeneity. Prognosis of these tumors relies largely on the extent of resection. Fluorescence guided surgery using 5-ALA as adjunct has been on the rise in the recent years. However, 5-ALA has been ineffective in a small subset of population with similar histological phenotypes but varying metabolic/biochemical properties. Visualized fluorescence can sometimes be subjective and lead to variability in defining fluorescing regions with respect to their biological grade. Objective assessment of fluorescence is possible using spectroscopic techniques and with ex vivo PpIX assessment assays. The biometric study in our previous work revealed that even with objective assessment using PpIX assays, there exists a small subpopulation of tumor cells with similar histological phenotypes but discordant metabolic/biochemical properties w.r.t accumulation of PpIX. In the current study, we extended the investigation further and have carried out proteomic analysis of high-grade glioma tissue samples resected using 5-ALA fluorescence guided surgery to understand molecular differences leading to differential fluorescence in these complex and heterogenous tumors.

INSTRUMENT(S): Orbitrap Fusion

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Brain

DISEASE(S): Glioblastoma

SUBMITTER: Saicharan Ghantasala

LAB HEAD: Prof. Sanjeeva Srivastava

PROVIDER: PXD039170 | Pride | 2023-07-20

REPOSITORIES: Pride

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Project description:The metabolic impact of the common peroxisome proliferator-activated receptor gamma isoform 2 (PPARÎ³2) variant Pro12Ala in human populations has been widely debated. We demonstrate using a Pro12Ala knock-in model that on chow Ala/Ala mice are leaner, have improved insulin sensitivity and plasma lipid profiles, and longer lifespan. Gene-environment interactions played a key role as high-fat feeding eliminated the beneficial effects of the Pro12Ala variant on adiposity, plasma lipids, and insulin sensitivity. The underlying molecular mechanisms involve changes in cofactor interaction and adiponectin signaling. Altogether, our results establish the Pro12Ala variant of PparÎ³2 as an important modulator in metabolic control that strongly depends on the metabolic context. PparÎ³ Pro12Ala and wild-type littermates were fed regular rodent chow or high-fat diet (D12330, 5560 kcal/kg, Research Diets, New Brunswick, NJ), as indicated. Only males were used to minimize possible effects of variation in estrus status of females. Since the body weight development of Pro/Ala heterozygote mice was intermediate to that of Pro/Pro and Ala/Ala mice, we focused on the two homozygote genotypes in further experimentation. Mice were housed with a 12h light-dark cycle and had free access to water and food. Eight week old male Pro/Pro and Ala/Ala littermate mice were fed either regular rodent chow or high-fat diet (as above) for 15 weeks (n = 8-10 per group), at which time blood and tissue samples were collected. Total RNA extraction, sample amplification, labeling, and microarray (Genechip Mouse Genome 430 2.0 Array, Affymetrix Inc., Santa Clara, CA) processing steps were performed by the Rosetta Inpharmatics Gene Expression Laboratory (Seattle, WA) using custom automated procedures in compliance with manufacturer protocols. Microarray data was processed using Rosetta Resolver (Rosetta Inpharmatics, Seattle, WA) and expressed as relative to the virtual pool of HFD â Pro/Pro group (mlratio). Gene expression signatures were generated from these mlratios for expressed genes (25% of RMA processed intensities above 60% quantile) by T-testing (Pro/Pro vs. Ala/Ala) and correcting for multiple testing (20% cut-off for False Discovery Rate). GSEA was performed according to provider suggestions, Pro/Pro vs. Ala/Ala. Statistical significance was declared if P < 0.05.

			Action	DRS
	ALA_Protein_groups.rar	Other
	ALA_metadata.xlsx	Xlsx
	Human_Proteome_May2022.fasta	Fasta
	P10_F_1_1.raw	Raw
	P10_F_1_2.raw	Raw

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Proteomics analysis of High-grade glioma tissues showing differential fluorescence following 5-Aminolevulinic acid guided surgery

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