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PBMC proteomics in paediatric obesity

ABSTRACT: We isolated cell lysate proteins from PBMCs obtained from 71 children and adolescents (age 5-18) with normal weight, overweight or obesity. Body composition was measured using bioelectrical impedance analysis. Nano-electrospray liquid chromatography-tandem mass spectrometry was used to measure protein abundances. Controlling for patient sex, we investigated changes in abundance depending on leukocyte counts and on percentage body fat (PBF). We show protein expression in PBMCs is altered in children with increased PBF, especially in mitochondrial proteins. These obesity-associated changes in are better explained by PBF than by purely height and weight-based metrics, supporting the utility of bioelectrical impedance analysis in obesity research.

INSTRUMENT(S): Q Exactive Plus

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Blood Cell, Blood

DISEASE(S): Obesity

SUBMITTER: Maya Petek

LAB HEAD: Uroš Potočnik

PROVIDER: PXD064634 | Pride | 2025-07-05

REPOSITORIES: Pride

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Project description:Background: The mechanisms of how genetic variants (SNPs) identified in genome-wide association studies act to influence body mass remain unknown for most of these SNPs, which continue to puzzle the scientific community. Recent evidence points to epigenetic and chromatin state of the genome to have an important role. Methods: 355 healthy young individuals were genotyped for 52 known obesity-associated SNPs and we obtained DNA methylation levels in their blood using the Illumina 450K BeadChip. Associations between alleles and methylation at proximal cytosine residues were tested using a linear model adjusted for age, sex, weight category and a proxy for blood cell type counts. For replication in other tissues, we used two open-access datasets (skin fibroblasts, n=62; four brain regions, n=121-133) and an additional dataset in subcutaneous and visceral fat (n=149). Results: We found that alleles at 28 of these obesity-associated SNPs associate with methylation levels at 107 proximal CpG sites. Out of 107 CpG sites, 38 are located in gene promoters, including genes strongly implicated in obesity (MIR148A, BDNF, PTPMT1, NR1H3, MGAT1, SCGB3A1, HOXC12, PMAIP1, PSIP1, RPS10-NUDT3, RPS10, SKOR1, MAP2K5, SIX5, AGRN, IMMP1L, ELP4, ITIH4, SEMA3G, POMC, ADCY3, SSPN, LGR4, TUFM, MIR4721, SULT1A1, SULT1A2, APOBR, CLN3, SPNS1,SH2B1, ATXN2L, and IL27). Interestingly, the associated SNPs are in known eQTLs for some of these genes. We also found that the 107 CpGs are enriched in enhancers in peripheral blood mononuclear cells. Finally, our results indicate that some of these associations are not be blood-specific as we successfully replicated four associations in skin fibroblasts. Conclusions: Our results strongly suggest that many obesity-associated SNPs are associated with proximal gene regulation, which was reflected by association of obesity risk allele genotypes with differential DNA methylation. This study highlights the importance of DNA methylation and other chromatin marks as a way to understand the molecular basis of genetic variants associated to human diseases and traits. Bisulphite converted DNA from 355 individuals aged 14-34 were hybridised to the Illumina Infinium 450k Human Methylation Beadchip.

			Action	DRS
	LBP_052.msf	Msf
	LBP_052.raw	Raw
	LBP_053.msf	Msf
	LBP_053.raw	Raw
	LBP_054.msf	Msf

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PBMC proteomics in paediatric obesity

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