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Rapid and reproducible characterization of sickling during automated deoxygenation in sickle cell disease patients.
Am J Hematol. 2019 05; 94(5):575-584.AJ

Abstract

In sickle cell disease (SCD), sickle hemoglobin (HbS) polymerizes upon deoxygenation, resulting in sickling of red blood cells (RBCs). These sickled RBCs have strongly reduced deformability, leading to vaso-occlusive crises and chronic hemolytic anemia. To date, there are no reliable laboratory parameters or assays capable of predicting disease severity or monitoring treatment effects. We here report on the oxygenscan, a newly developed method to measure RBC deformability (expressed as Elongation Index - EI) as a function of pO2 . Upon a standardized, 22 minute, automated cycle of deoxygenation (pO2 median 16 mmHg ± 0.17) and reoxygenation, a number of clinically relevant parameters are produced in a highly reproducible manner (coefficients of variation <5%). In particular, physiological modulators of oxygen affinity, such as, pH and 2,3-diphosphoglycerate showed a significant correlation (respectively R = -0.993 and R = 0.980) with Point of Sickling (PoS5%), which is defined as the pO2 where a 5% decrease in EI is observed during deoxygenation. Furthermore, in vitro treatment with antisickling agents, including GBT440, which alter the oxygen affinity of hemoglobin, caused a reproducible left-shift of the PoS, indicating improved deformability at lower oxygen tensions. When RBCs from 21 SCD patients were analyzed, we observed a significantly higher PoS in untreated homozygous SCD patients compared to treated patients and other genotypes. We conclude that the oxygenscan is a state-of-the-art technique that allows for rapid analysis of sickling behavior in SCD patients. The method is promising for personalized treatment, development of new treatment strategies and could have potential in prediction of complications.

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Authors+Show Affiliations

Rab MAE
Laboratory of Clinical Chemistry & Hematology, University Medical Center Utrecht, Utrecht University, Utrecht, The Netherlands. Van Creveldkliniek, University Medical Center Utrecht, Utrecht University, Utrecht, The Netherlands.
van Oirschot BA
Laboratory of Clinical Chemistry & Hematology, University Medical Center Utrecht, Utrecht University, Utrecht, The Netherlands.
Bos J
Laboratory of Clinical Chemistry & Hematology, University Medical Center Utrecht, Utrecht University, Utrecht, The Netherlands.
Merkx TH
Laboratory of Clinical Chemistry & Hematology, University Medical Center Utrecht, Utrecht University, Utrecht, The Netherlands.
van Wesel ACW
Laboratory of Clinical Chemistry & Hematology, University Medical Center Utrecht, Utrecht University, Utrecht, The Netherlands.
Abdulmalik O
Division of Hematology, The Children's Hospital of Philadelphia, Philadelphia, Pennsylvania.
Safo MK
Department of Medicinal Chemistry, Institute for Structural Biology, Drug Discovery and Development, School of Pharmacy, Virginia Commonwealth University, Virginia.
Versluijs BA
Department of Pediatric Hematology, University Medical Center Utrecht, Utrecht University, Utrecht, The Netherlands.
Houwing ME
Department of Pediatric Hematology, Erasmus University Medical Center- Sophia Children's Hospital, Rotterdam, The Netherlands.
Cnossen MH
Department of Pediatric Hematology, Erasmus University Medical Center- Sophia Children's Hospital, Rotterdam, The Netherlands.
Riedl J
Result Laboratory, Albert Schweitzer Hospital, Dordrecht, The Netherlands.
Schutgens REG
Van Creveldkliniek, University Medical Center Utrecht, Utrecht University, Utrecht, The Netherlands.
Pasterkamp G
Laboratory of Clinical Chemistry & Hematology, University Medical Center Utrecht, Utrecht University, Utrecht, The Netherlands.
Bartels M
Department of Pediatric Hematology, University Medical Center Utrecht, Utrecht University, Utrecht, The Netherlands.
van Beers EJ
Van Creveldkliniek, University Medical Center Utrecht, Utrecht University, Utrecht, The Netherlands.
van Wijk R
Laboratory of Clinical Chemistry & Hematology, University Medical Center Utrecht, Utrecht University, Utrecht, The Netherlands.

MeSH

Anemia, Sickle CellBenzaldehydesErythrocytes, AbnormalHemoglobin, SickleHumansOxygenPyrazinesPyrazoles

Pub Type(s)

Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

30784099

Citation

Rab, Minke A E., et al. "Rapid and Reproducible Characterization of Sickling During Automated Deoxygenation in Sickle Cell Disease Patients." American Journal of Hematology, vol. 94, no. 5, 2019, pp. 575-584.
Rab MAE, van Oirschot BA, Bos J, et al. Rapid and reproducible characterization of sickling during automated deoxygenation in sickle cell disease patients. Am J Hematol. 2019;94(5):575-584.
Rab, M. A. E., van Oirschot, B. A., Bos, J., Merkx, T. H., van Wesel, A. C. W., Abdulmalik, O., Safo, M. K., Versluijs, B. A., Houwing, M. E., Cnossen, M. H., Riedl, J., Schutgens, R. E. G., Pasterkamp, G., Bartels, M., van Beers, E. J., & van Wijk, R. (2019). Rapid and reproducible characterization of sickling during automated deoxygenation in sickle cell disease patients. American Journal of Hematology, 94(5), 575-584. https://doi.org/10.1002/ajh.25443
Rab MAE, et al. Rapid and Reproducible Characterization of Sickling During Automated Deoxygenation in Sickle Cell Disease Patients. Am J Hematol. 2019;94(5):575-584. PubMed PMID: 30784099.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Rapid and reproducible characterization of sickling during automated deoxygenation in sickle cell disease patients. AU - Rab,Minke A E, AU - van Oirschot,Brigitte A, AU - Bos,Jennifer, AU - Merkx,Tesy H, AU - van Wesel,Annet C W, AU - Abdulmalik,Osheiza, AU - Safo,Martin K, AU - Versluijs,Birgitta A, AU - Houwing,Maite E, AU - Cnossen,Marjon H, AU - Riedl,Jurgen, AU - Schutgens,Roger E G, AU - Pasterkamp,Gerard, AU - Bartels,Marije, AU - van Beers,Eduard J, AU - van Wijk,Richard, Y1 - 2019/03/08/ PY - 2019/02/11/received PY - 2019/02/17/revised PY - 2019/02/19/accepted PY - 2019/2/21/pubmed PY - 2020/1/28/medline PY - 2019/2/21/entrez SP - 575 EP - 584 JF - American journal of hematology JO - Am J Hematol VL - 94 IS - 5 N2 - In sickle cell disease (SCD), sickle hemoglobin (HbS) polymerizes upon deoxygenation, resulting in sickling of red blood cells (RBCs). These sickled RBCs have strongly reduced deformability, leading to vaso-occlusive crises and chronic hemolytic anemia. To date, there are no reliable laboratory parameters or assays capable of predicting disease severity or monitoring treatment effects. We here report on the oxygenscan, a newly developed method to measure RBC deformability (expressed as Elongation Index - EI) as a function of pO2 . Upon a standardized, 22 minute, automated cycle of deoxygenation (pO2 median 16 mmHg ± 0.17) and reoxygenation, a number of clinically relevant parameters are produced in a highly reproducible manner (coefficients of variation <5%). In particular, physiological modulators of oxygen affinity, such as, pH and 2,3-diphosphoglycerate showed a significant correlation (respectively R = -0.993 and R = 0.980) with Point of Sickling (PoS5%), which is defined as the pO2 where a 5% decrease in EI is observed during deoxygenation. Furthermore, in vitro treatment with antisickling agents, including GBT440, which alter the oxygen affinity of hemoglobin, caused a reproducible left-shift of the PoS, indicating improved deformability at lower oxygen tensions. When RBCs from 21 SCD patients were analyzed, we observed a significantly higher PoS in untreated homozygous SCD patients compared to treated patients and other genotypes. We conclude that the oxygenscan is a state-of-the-art technique that allows for rapid analysis of sickling behavior in SCD patients. The method is promising for personalized treatment, development of new treatment strategies and could have potential in prediction of complications. SN - 1096-8652 UR - https://www.unboundmedicine.com/medline/citation/30784099/Rapid_and_reproducible_characterization_of_sickling_during_automated_deoxygenation_in_sickle_cell_disease_patients_ DB - PRIME DP - Unbound Medicine ER -