Red Blood Cell- Omics

The NHLBI Recipient Epidemiology and Donor Evaluation Study (REDS-III) Red Blood Cell Omics (RBC-Omics) Study

The REDS-III RBC-Omics research team focused on a simple observation, that the variability in donor RBC stability and function during cold storage and post-transfusion recovery is more dependent on the donor than the time the unit is in storage. For example, mouse and human female blood is more stable during storage than male blood, and RBC transfusions from donors with common mutations like sickle cell trait (HbAS) exhibit enhanced storage hemolysis and lower post-transfusion recovery. These observations informed our overarching hypothesis that evolved variability in genes encoding RBC proteins significantly modulate RBC storage properties and function and recovery after transfusion. This genetic variability has evolved in large part as a consequence of endemic malaria, which has exerted evolutionary pressure on the human genome. While it is well appreciated that this variability is significant in admixed populations, especially African American, Asian, Hispanic, Arabic, Indian and Mediterranean populations, no previous studies have explored how common and rare variants might modulate donor RBCs during storage and post-transfusion, nor how these variants might affect susceptibility to hemolysis in human diseases, like sickle cell anemia.

To address this question, the REDS-III RBC-Omics Study consented and enrolled 13,403 RBC donors through the efforts of the four “hub” blood centers in the U.S, with over-recruitment of racial-ethnic minority and high-frequency donors. The study team developed and applied high throughput automated assays to measure various types of hemolysis, including osmotic, oxidative and spontaneous storage hemolysis, in samples of the leukocyte-reduced RBC (LR-RBC) components that had been stored under blood bank conditions for 42 days. Recall of 664 of the enrolled donors enabled confirmation of “heritability” of hemolysis phenotypes and detailed analyses of kinetics of hemolysis parameters over storage of LR-RBC and corresponding changes in RBC metabolomic profiles.  We then performed the largest genome-wide association study (GWAS) study of healthy human blood donors to date using a custom Transfusion Medicine Array, comparing high density single nucleotide polymorphisms (SNPs) with RBC responses to canonical in vitro stressors following 4°C storage for 42 days as well as to metabolomic findings. These RBC-Omics studies have identified candidate SNPs at genome wide significance that are associated with enhanced or reduced responses to hemolytic perturbations. 

We plan to continue to explore the function of these common and rare variant SNPs in vitro by correlating genetic and RBC structural and metabolomic findings, and in vivo, both in mouse models of transfusion and also in humans with measurements such as post-transfusion RBC recovery and function in recipient of RBC-Omics donors in the REDS-III linked donor recipient database and autologous biotin labeled transfusion-recovery studies focused on selected recalled RBC-Omics donors. We anticipate that these findings will  contribute to a Precision Transfusion Medicine field, in which RBC donor genotype determines the time limits of RBC storage and helps predicts in vivo RBC recovery and survival. We also anticipate that the SNPs that we identify that correlate with in vitro hemolysis, as well as other associations like age and gender, will predict risk of hemolysis in human diseases like sickle cell anemia.

Translating red cell “omics” into new perspectives in transfusion medicine: mining the gems in data Mountains

S.L. Spitalnik and D.V. Devine, pages 2-5

Blood, sweat, and tears: Red Blood Cell‐Omics study objectives, design, and recruitment activities

S.M. Endres-Dighe, et al, pages 46-56

Piloting and implementation of quality assessment and quality control procedures in RBC‐Omics: a large multi‐center study of red blood cell hemolysis during storage

M. Stone, et al, pages 57-66

Frequent blood donations alter susceptibility of red blood cells to storage‐ and stress‐induced hemolysis

T. Kanias, et al, pages 67-78

Intradonor reproducibility and changes in hemolytic variables during red blood cell storage: results of recall phase of the REDS‐III RBC‐Omics study

M.C. Lanteri, et al, pages 79-88

Heterogeneity of blood processing and storage additives in different centers impacts stored red blood cell metabolism as much as storage time: lessons from REDS‐III—Omics

A. D’Alessandro, et al, pages 89-100

Development and evaluation of a transfusion medicine genome wide genotyping array

Y. Guo, et al , pages 101- 111

Methylation of protein aspartates and deamidated asparagines as a function of blood bank storage and oxidative stress in human red blood cells

J. Reisz, et al