Assessing the kinetics of oxygen-unloading from red cells using FlowScore, a flow-cytometric proxy of the functional quality of blood

Julija Rabcuka; Peter A. Smethurst; Katharina Dammert; Jarob Saker; Gemma Aran; Geraldine M. Walsh; Joanne C.G. Tan; Margarita Codinach; Ken McTaggart; Denese C. Marks; Stephan J.L. Bakker; Amy McMahon; Emanuele Di Angelantonio; David J. Roberts; Slawomir Blonski; Piotr M. Korczyk; Atsushi Shirakami; Rebecca Cardigan; Pawel Swietach

doi:10.1016/j.ebiom.2024.105498

Assessing the kinetics of oxygen-unloading from red cells using FlowScore, a flow-cytometric proxy of the functional quality of blood

Julija Rabcuka, Peter A. Smethurst, Katharina Dammert, Jarob Saker, Gemma Aran, Geraldine M. Walsh, Joanne C.G. Tan, Margarita Codinach, Ken McTaggart, Denese C. Marks, Stephan J.L. Bakker, Amy McMahon, Emanuele Di Angelantonio, David J. Roberts, Slawomir Blonski, Piotr M. Korczyk, Atsushi Shirakami, Rebecca Cardigan, Pawel Swietach^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

Abstract

Background: Metrics evaluating the functional quality of red blood cells (RBCs) must consider their role in oxygen delivery. Whereas oxygen-carrying capacity is routinely reported using haemoglobin assays, the rate of oxygen exchange is not measured, yet also important for tissue oxygenation. Since oxygen-unloading depends on the diffusion pathlength inside RBCs, cell geometry offers a plausible surrogate. Methods: We related the time-constant of oxygen-unloading (τ), measured using single-cell oxygen saturation imaging, with flow-cytometric variables recorded on a haematology analyser. Experiments compared freshly-drawn RBCs with stored RBCs, wherein metabolic run-down and spherical remodelling hinder oxygen unloading. Findings: Multivariable regression related τ to a ratio of side- and forward-scatter, referred to herein as FlowScore. FlowScore was able to distinguish, with sensitivity and specificity >80%, freshly drawn blood from blood that underwent storage-related kinetic attrition in O₂-handling. Moreover, FlowScore predicted τ restoration upon biochemical rejuvenation of stored blood. Since RBC geometry and metabolic state are related, variants of FlowScore estimated [ATP] and [2,3-diphosphoglycerate]. The veracity of FlowScore was confirmed by four blood-banking systems (Australia, Canada, England, Spain). Applying FlowScore to data from the COMPARE study revealed a positive association with the time-delay from sample collection to measurement, which was verified experimentally. The LifeLines dataset revealed age, sex, and smoking among factors affecting FlowScore. Interpretation: We establish FlowScore as a widely-accessible and cost-effective surrogate of RBC oxygen-unloading kinetics. As a metric of a cellular process that is sensitive to storage and disease, we propose FlowScore as an RBC quality marker for blood-banking and haematology. Funding: See Acknowledgements.

Original language	English
Article number	105498
Journal	EBioMedicine
Volume	111
DOIs	https://doi.org/10.1016/j.ebiom.2024.105498
Publication status	Published - Jan 2025
Externally published	Yes

Bibliographical note

Publisher Copyright:
© 2024 The Authors

Keywords

Assay
Erythrocytes
Haematology
Oxygen transport
Storage lesion
Transfusion

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10.1016/j.ebiom.2024.105498

Cite this

Rabcuka, J., Smethurst, P. A., Dammert, K., Saker, J., Aran, G., Walsh, G. M., Tan, J. C. G., Codinach, M., McTaggart, K., Marks, D. C., Bakker, S. J. L., McMahon, A., Di Angelantonio, E., Roberts, D. J., Blonski, S., Korczyk, P. M., Shirakami, A., Cardigan, R., & Swietach, P. (2025). Assessing the kinetics of oxygen-unloading from red cells using FlowScore, a flow-cytometric proxy of the functional quality of blood. EBioMedicine, 111, Article 105498. https://doi.org/10.1016/j.ebiom.2024.105498

@article{20d249337c144c65bcd18cbe426218a7,

title = "Assessing the kinetics of oxygen-unloading from red cells using FlowScore, a flow-cytometric proxy of the functional quality of blood",

abstract = "Background: Metrics evaluating the functional quality of red blood cells (RBCs) must consider their role in oxygen delivery. Whereas oxygen-carrying capacity is routinely reported using haemoglobin assays, the rate of oxygen exchange is not measured, yet also important for tissue oxygenation. Since oxygen-unloading depends on the diffusion pathlength inside RBCs, cell geometry offers a plausible surrogate. Methods: We related the time-constant of oxygen-unloading (τ), measured using single-cell oxygen saturation imaging, with flow-cytometric variables recorded on a haematology analyser. Experiments compared freshly-drawn RBCs with stored RBCs, wherein metabolic run-down and spherical remodelling hinder oxygen unloading. Findings: Multivariable regression related τ to a ratio of side- and forward-scatter, referred to herein as FlowScore. FlowScore was able to distinguish, with sensitivity and specificity >80%, freshly drawn blood from blood that underwent storage-related kinetic attrition in O2-handling. Moreover, FlowScore predicted τ restoration upon biochemical rejuvenation of stored blood. Since RBC geometry and metabolic state are related, variants of FlowScore estimated [ATP] and [2,3-diphosphoglycerate]. The veracity of FlowScore was confirmed by four blood-banking systems (Australia, Canada, England, Spain). Applying FlowScore to data from the COMPARE study revealed a positive association with the time-delay from sample collection to measurement, which was verified experimentally. The LifeLines dataset revealed age, sex, and smoking among factors affecting FlowScore. Interpretation: We establish FlowScore as a widely-accessible and cost-effective surrogate of RBC oxygen-unloading kinetics. As a metric of a cellular process that is sensitive to storage and disease, we propose FlowScore as an RBC quality marker for blood-banking and haematology. Funding: See Acknowledgements.",

keywords = "Assay, Erythrocytes, Haematology, Oxygen transport, Storage lesion, Transfusion",

author = "Julija Rabcuka and Smethurst, {Peter A.} and Katharina Dammert and Jarob Saker and Gemma Aran and Walsh, {Geraldine M.} and Tan, {Joanne C.G.} and Margarita Codinach and Ken McTaggart and Marks, {Denese C.} and Bakker, {Stephan J.L.} and Amy McMahon and {Di Angelantonio}, Emanuele and Roberts, {David J.} and Slawomir Blonski and Korczyk, {Piotr M.} and Atsushi Shirakami and Rebecca Cardigan and Pawel Swietach",

note = "Publisher Copyright: {\textcopyright} 2024 The Authors",

year = "2025",

month = jan,

doi = "10.1016/j.ebiom.2024.105498",

language = "English",

volume = "111",

journal = "EBioMedicine",

issn = "2352-3964",

publisher = "Elsevier B.V.",

}

Rabcuka, J, Smethurst, PA, Dammert, K, Saker, J, Aran, G, Walsh, GM, Tan, JCG, Codinach, M, McTaggart, K, Marks, DC, Bakker, SJL, McMahon, A, Di Angelantonio, E, Roberts, DJ, Blonski, S, Korczyk, PM, Shirakami, A, Cardigan, R & Swietach, P 2025, 'Assessing the kinetics of oxygen-unloading from red cells using FlowScore, a flow-cytometric proxy of the functional quality of blood', EBioMedicine, vol. 111, 105498. https://doi.org/10.1016/j.ebiom.2024.105498

TY - JOUR

T1 - Assessing the kinetics of oxygen-unloading from red cells using FlowScore, a flow-cytometric proxy of the functional quality of blood

AU - Rabcuka, Julija

AU - Smethurst, Peter A.

AU - Dammert, Katharina

AU - Saker, Jarob

AU - Aran, Gemma

AU - Walsh, Geraldine M.

AU - Tan, Joanne C.G.

AU - Codinach, Margarita

AU - McTaggart, Ken

AU - Marks, Denese C.

AU - Bakker, Stephan J.L.

AU - McMahon, Amy

AU - Di Angelantonio, Emanuele

AU - Roberts, David J.

AU - Blonski, Slawomir

AU - Korczyk, Piotr M.

AU - Shirakami, Atsushi

AU - Cardigan, Rebecca

AU - Swietach, Pawel

PY - 2025/1

Y1 - 2025/1

N2 - Background: Metrics evaluating the functional quality of red blood cells (RBCs) must consider their role in oxygen delivery. Whereas oxygen-carrying capacity is routinely reported using haemoglobin assays, the rate of oxygen exchange is not measured, yet also important for tissue oxygenation. Since oxygen-unloading depends on the diffusion pathlength inside RBCs, cell geometry offers a plausible surrogate. Methods: We related the time-constant of oxygen-unloading (τ), measured using single-cell oxygen saturation imaging, with flow-cytometric variables recorded on a haematology analyser. Experiments compared freshly-drawn RBCs with stored RBCs, wherein metabolic run-down and spherical remodelling hinder oxygen unloading. Findings: Multivariable regression related τ to a ratio of side- and forward-scatter, referred to herein as FlowScore. FlowScore was able to distinguish, with sensitivity and specificity >80%, freshly drawn blood from blood that underwent storage-related kinetic attrition in O2-handling. Moreover, FlowScore predicted τ restoration upon biochemical rejuvenation of stored blood. Since RBC geometry and metabolic state are related, variants of FlowScore estimated [ATP] and [2,3-diphosphoglycerate]. The veracity of FlowScore was confirmed by four blood-banking systems (Australia, Canada, England, Spain). Applying FlowScore to data from the COMPARE study revealed a positive association with the time-delay from sample collection to measurement, which was verified experimentally. The LifeLines dataset revealed age, sex, and smoking among factors affecting FlowScore. Interpretation: We establish FlowScore as a widely-accessible and cost-effective surrogate of RBC oxygen-unloading kinetics. As a metric of a cellular process that is sensitive to storage and disease, we propose FlowScore as an RBC quality marker for blood-banking and haematology. Funding: See Acknowledgements.

AB - Background: Metrics evaluating the functional quality of red blood cells (RBCs) must consider their role in oxygen delivery. Whereas oxygen-carrying capacity is routinely reported using haemoglobin assays, the rate of oxygen exchange is not measured, yet also important for tissue oxygenation. Since oxygen-unloading depends on the diffusion pathlength inside RBCs, cell geometry offers a plausible surrogate. Methods: We related the time-constant of oxygen-unloading (τ), measured using single-cell oxygen saturation imaging, with flow-cytometric variables recorded on a haematology analyser. Experiments compared freshly-drawn RBCs with stored RBCs, wherein metabolic run-down and spherical remodelling hinder oxygen unloading. Findings: Multivariable regression related τ to a ratio of side- and forward-scatter, referred to herein as FlowScore. FlowScore was able to distinguish, with sensitivity and specificity >80%, freshly drawn blood from blood that underwent storage-related kinetic attrition in O2-handling. Moreover, FlowScore predicted τ restoration upon biochemical rejuvenation of stored blood. Since RBC geometry and metabolic state are related, variants of FlowScore estimated [ATP] and [2,3-diphosphoglycerate]. The veracity of FlowScore was confirmed by four blood-banking systems (Australia, Canada, England, Spain). Applying FlowScore to data from the COMPARE study revealed a positive association with the time-delay from sample collection to measurement, which was verified experimentally. The LifeLines dataset revealed age, sex, and smoking among factors affecting FlowScore. Interpretation: We establish FlowScore as a widely-accessible and cost-effective surrogate of RBC oxygen-unloading kinetics. As a metric of a cellular process that is sensitive to storage and disease, we propose FlowScore as an RBC quality marker for blood-banking and haematology. Funding: See Acknowledgements.

KW - Assay

KW - Erythrocytes

KW - Haematology

KW - Oxygen transport

KW - Storage lesion

KW - Transfusion

UR - http://www.scopus.com/inward/record.url?scp=85211585335&partnerID=8YFLogxK

U2 - 10.1016/j.ebiom.2024.105498

DO - 10.1016/j.ebiom.2024.105498

M3 - Article

AN - SCOPUS:85211585335

SN - 2352-3964

VL - 111

JO - EBioMedicine

JF - EBioMedicine

M1 - 105498

ER -

Assessing the kinetics of oxygen-unloading from red cells using FlowScore, a flow-cytometric proxy of the functional quality of blood

Abstract

Bibliographical note

Keywords

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