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Mass spectrometry-based analysis on the impact of iron status and whole blood donation on the global plasma proteome

ABSTRACT: Blood donation is generally considered to be safe, however to ensure blood donor health it is important to monitor the impact of a whole blood donation. Study design and methods Here, we used quantitative mass spectrometry to assess the global plasma proteomic impact of longitudinal whole blood donation in 5 new and 4 regular donors over a period of 180 days as well as the effect on iron status, by selecting 78 donors based on high levels of ferritin and hemoglobin. The majority of plasma proteins were stable between whole blood donors and demonstrated a low coefficient of variation, irrespective to iron status or donors status (new or regular). Longitudinal analysis showed limited differences between the plasma proteomes of regular and new upon a whole blood donation, apart from a consistent and significant short-term decrease in fibronectin. Plasma protein levels measured with MS highly correlated with blood count and lipid parameters, validating our MS-approach. Based on protein co-expression, we observed protein complexes of diverse biological processes, platelet and neutrophil signatures, complement and immune responses with a highly correlating cluster of IGHM, IGJ and CD5L. We showed higher inter- than intra-individual variation, most notably of immunoglobulins and allelic variants for alpha-1 antitrypsin. Overall, this study shows that whole blood donation has limited impact for both new and regular donors apart from short-term decreases in fibronectin and that varying iron statuses do not affect the global plasma proteome. We showed that individual specific signatures were reflected on the plasma proteome as well as detection of allelic variants

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Blood Plasma

SUBMITTER: Arie Hoogendijk

LAB HEAD: M van den Biggelaar

PROVIDER: PXD035447 | Pride | 2023-02-01

REPOSITORIES: Pride

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Project description:<p>The Susan G. Komen for the Cure® Tissue Bank at the IU Simon Cancer Center [<a href="www.komentissuebank.iu.edu" target="_blank">www.komentissuebank.iu.edu</a>] (KTB) was established expressly for the prospective collection of normal, healthy breast tissue from volunteer donors. Blood is also obtained from donors at the time of donation and is processed for serum, plasma and peripheral blood leukocyte DNA. Specimens are annotated and data includes age, race, ethnicity, personal health history, family cancer history, medication usage at the time of donation, and breast cancer risk factors.</p> <p>Premenopausal donors to the KTB were identified by a query of the Bank's database. Hematoxylin and eosin stained sections of the formalin-fixed paraffin-embedded tissue of the identified donors were reviewed and tissue was graded on the basis of the abundance of epithelium within the section. Only tissue containing abundant epithelium was considered for this study. Based on dates, the specimens of nine women in the follicular phase of the menstrual cycle and five in the luteal phase were chosen. Six donors using hormonal contraception at the time of donation were also included. Whole blood obtained from 19 of the 20 donors at the time of tissue donation was available and it was processed for serum. Estradiol, estriol, luteinizing hormone and progesterone concentrations were determined by the Indiana University Health Pathology Laboratory using a Beckman Unicel DxI 800 Immunoassay System. The phase of the menstrual cycle was verified by serum progesterone concentration. The epithelium of these 20 specimens was microdissected from multiple 8 micron frozen tissue sections. Total RNA extracted from the tissue was subsequently depleted of rRNA via locked nucleic acid probes. This enabled profiling of both poly-A and non-poly-A RNA species. Barcoded cDNA libraries from the 20 normal breast epithelia were prepared and sequenced on an Applied Biosystems (AB) SOLiD3 or SOLiD 4 platform. RNA-seq reads for each sample were then mapped to the human genome (hg19) using the LifeScope software version 2.5.1 (Life Technologies, Foster City, CA) and BAM (Binary Alignment/Map) files generated. Read counts for each gene were derived from the output BAM files using the RefSeq database (UCSC Genome Brower) as the gene model.</p>

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Mass spectrometry-based analysis on the impact of iron status and whole blood donation on the global plasma proteome

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