GPX4 status affects GPX4 protein and peptide levels, and the omics storage lesion in REDS recalled donors. Of the 13 091 index donors, 643 were invited to donate a second unit of blood (recalled cohort) based on their extreme hemolytic propensity (ie, donors who ranked <5th or >95th percentile in the index cohort (A). These second units were stored for 42 days, and tested at storage day 10, 23, and 42 for omics and hemolytic parameters. (B) Uniform manifold approximation and projection analysis of omics data by storage duration shows that donors carrying 1 or 2 rs73507255 alternative alleles (missense mutation) clustered separately from the rest of the donor population. (C-D) Lower oxidative and osmotic hemolysis were measured in RBCs from donors carrying 2 alleles of the rs8178977 SNP, which results in higher levels of GPX4 protein (E). Indeed, while GPX4 protein levels were not affected by storage duration (F), donors carrying 1 or 2 copies of the rs8178977 alleles displayed higher levels of GPX4 throughout storage (G), especially in younger donors (H). Strong associations between GPX4 protein and peptide levels were observed (I), especially for free or glutathionylated (Cys) peptides in donors carrying the rs8178973 and rs8178977 SNPs (J-M).
Figure 3.

GPX4 status affects GPX4 protein and peptide levels, and the omics storage lesion in REDS recalled donors. Of the 13 091 index donors, 643 were invited to donate a second unit of blood (recalled cohort) based on their extreme hemolytic propensity (ie, donors who ranked <5th or >95th percentile in the index cohort (A). These second units were stored for 42 days, and tested at storage day 10, 23, and 42 for omics and hemolytic parameters. (B) Uniform manifold approximation and projection analysis of omics data by storage duration shows that donors carrying 1 or 2 rs73507255 alternative alleles (missense mutation) clustered separately from the rest of the donor population. (C-D) Lower oxidative and osmotic hemolysis were measured in RBCs from donors carrying 2 alleles of the rs8178977 SNP, which results in higher levels of GPX4 protein (E). Indeed, while GPX4 protein levels were not affected by storage duration (F), donors carrying 1 or 2 copies of the rs8178977 alleles displayed higher levels of GPX4 throughout storage (G), especially in younger donors (H). Strong associations between GPX4 protein and peptide levels were observed (I), especially for free or glutathionylated (Cys) peptides in donors carrying the rs8178973 and rs8178977 SNPs (J-M).

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