Regling K, DO , Cashen K , Gadgeel M, Xi Y, Herppich A,Tarasev M, Hines P, Chitlur M, Blood (2020) 136 (Supplement 1): 14.
Bleeding and thrombosis remain the primary complications related to the use of extracorporeal membrane oxygenation (ECMO). To date, no single test or parameter has been identified to accurately predict the risk of these hemostatic complications. Thrombin generation may be the key marker for both thrombosis and bleeding, and may be influenced by inflammation. The aims of this study were to evaluate if novel laboratory tests including thrombin generation assay, neutrophil extracellular traps (NETs), microparticles (MPs), and red blood cell (RBC) membrane fragility/adhesion could better predict patients at risk for bleeding and/or thrombosis compared to routine laboratory tests. We hypothesized that ECMO related thrombosis is associated with increased thrombin generation and ECMO related bleeding is associated with decreased thrombin generation.
White J, Moira L, Gao X, Tarasev M, Chakraborty S, Emanuele M, Hines PC. Can red blood cell function assays assess response to red cell-modifying therapies? Clin Hemorheol Microcirc. 2021 Jan 7. doi: 10.3233/CH-200944. Epub ahead of print. PMID: 33459699.
Red blood cell (RBC)-modifying therapies have provided new opportunities for patients with sickle cell disease, although the absence of validated biomarkers of RBC function is a barrier to FDA approval and clinical adoption. Flow Adhesion (FA) and Mechanical Fragility (MF) biomarkers objectively stratify individuals with SCD into pro-adhesive vs pro-hemolytic phenotypes respectively, which may potentially help predict therapeutic responses.
Pittman DD, Hines PC, Beidler D, Rybin D, Frelinger AL, Michelson AD, Liu K, Gao X, White J, Zaidi AU, Charnigo RJ, Callaghan MU. Evaluation of Longitudinal Pain Study in Sickle Cell Disease (ELIPSIS) by patient-reported outcomes, actigraphy, and biomarkers. Blood. 2021 Apr 15;137(15):2010-2020. doi: 10.1182/blood.2020006020. PMID: 33067606; PMCID: PMC8057263.
Clinical trials in sickle cell disease (SCD) often focus on health care utilization for painful vaso-occlusive crises (VOCs). However, no objective, quantifiable pain biomarkers exist, pain is not specific to VOCs, health care utilization varies between patients, unreported at-home VOCs likely contribute to long-term outcomes, and patient-reported outcomes are seldom considered. This non interventional, longitudinal, 6-month study aimed to develop tools to identify VOCs in SCD patients with or without health care utilization.
Tarasev M, Chakraborty S, Light L, Alfano K, Pagani F.D. Red blood cell mechanical fragility as potential metric for assessing blood damage caused by implantable durable ventricular assist devices: Comparison of two types of centrifugal flow left ventricular assist devices, Progress in Pediatric Cardiology, Volume 56, 2020,101198, ISSN 1058-9813
Implantable Ventricular Assist Devices (VADs) have become a treatment of choice for patients with end-stage heart failure or cardiogenic shock, significantly increasing both survival rates and the quality of life of patients. Moreover, VAD use is growing as destination therapy for patients who require permanent mechanical cardiac circulatory support. This heightens the need to ensure VAD reliability and safety, even amidst challenges in optimization of pump design for minimal blood damage
Tarasev M, Muchnik M, Chakraborty S (2019) Impact of the Oscillating Bead Size and Shape
on Induced Mechanical Stress on Red Blood Cells and Associated Hemolysis in Bead Milling. Int J
Blood Res Disord 6:041. doi.org/10.23937/2469-5696/1410041
While in circulation, red blood cells (RBC) need to elastically undergo large deformations without lysing, an ability that may be compromised by cell membrane damage. Such can be tested in vitro by subjecting an RBC sample to external mechanical stress, e.g. through bead milling or oscillation of an object in a sample. In addition to controlling frequency and duration of oscillations, this approach can be further tailored by bead selection/design.
White J, Lindgren M, Liu K, Gao X, Jendeberg L, Hines P. Sevuparin blocks sickle blood cell adhesion and sickle-leucocyte rolling on immobilized L-selectin in a dose dependent manner. Br J Haematol. 2019 Mar;184(5):873-876. doi: 10.1111/bjh.15188. Epub 2018 May 16. PMID: 29767405.
Adhesion of sickle red blood cells (SSRBC) to the vascular endothelium may initiate and propagate vascular obstruction in sickle cell disease (SCD) (Hoover et al, 1979; Hebbel et al, 1980). Hebbel et al (1980) were the first to report a correlation between erythrocyte adherence and disease severity. Subsequent studies demonstrated that the pathological adhesion of SSRBCs involves red cell receptors, adhesive bridging proteins and endothelial receptors (Joneckis et al, 1993; Swerlick et al, 1993; Udani et al, 1998; Hillery et al, 2000). These complex and multimodal mechanisms of SSRBC adhesion may require a multi‐targeted approach to achieve the best clinical outcome.
Tarasev M, Muchnik M, Light L, Alfano K, Chakraborty S. Individual variability in response to a single sickling event for normal, sickle cell, and sickle trait erythrocytes. Transl Res. 2017 Mar;181:96-107. doi: 10.1016/j.trsl.2016.09.005. Epub 2016 Sep 23. PMID: 27728824.
Hemoglobin S (Hb-S) polymerization is the primary event in sickle cell disease causing irreversible damage to red blood cell (RBC) membranes over repeated polymerization cycles. A single polymerization triggered by a hypoxic environment was reported to result in reversibly (upon reoxygenation) decreased RBC deformability and increased mechanical fragility (MF). Individualized responses have not been reported, although RBC fragility can vary significantly even among healthy individuals. This study evaluates individual variability in response to a single hypoxia-induced sickling event, through changes in RBC MF.
Alfano KM, Chakraborty S, Tarasev M. Differences in bead-milling-induced hemolysis of red blood cells due to shape and size of oscillating bead. Biomed Mater Eng. 2016 Sep 28;27(4):405-412. doi: 10.3233/BME-161594. PMID: 27689573.
Red blood cell (RBC) susceptibility to hemolysis - or fragility - can be profiled by subjecting a sample to progressive durations of mechanical stress and measuring hemolysis upon each. The ability to control stress application with multiple variable parameters can be useful in various areas of research. Bead milling, by oscillating an object in a blood sample, can offer control of parameters including oscillation force and frequency.
Tarasev M, Chakraborty S, Light L, Davenport R. Impact of environment on Red Blood Cell ability to withstand mechanical stress. Clin Hemorheol Microcirc. 2016 Nov 4;64(1):21-33. doi: 10.3233/CH-152037. PMID: 26890109.
Susceptibility of red blood cells (RBC) to hemolysis under mechanical stress is represented by RBC mechanical fragility (MF), with different types or intensities of stress potentially emphasizing different perturbations of RBC membranes. RBC membrane mechanics were shown to depend on cell environment, with many details not yet understood.
Lancelot M, White J, Sarnaik S, Hines P. Low molecular weight heparin inhibits sickle erythrocyte adhesion to VCAM-1 through VLA-4 blockade in a standardized microfluidic flow adhesion assay. Br J Haematol. 2017 Aug;178(3):479-481. doi: 10.1111/bjh.14137. Epub 2016 Jun 24. PMID: 27341635.
The vaso-occlusive events in sickle cell disease (SCD) begin in early childhood, warranting the need for more preventative and therapeutic interventions for those affected. Vaso-occlusion is partly caused by adhesion of sickle erythrocytes (SSRBCs) to components of the vascular wall and to circulating leucocytes (WBCs). SSRBCs have greater adhesion to the vascular endothelium and sub-endothelial matrix (SEM) compared to RBCs from unaffected individuals.
White J, Krishnamoorthy S, Gupta D, Lancelot M, Moore N, Sarnaik S, Hobbs WE 2nd, Light DR, Hines P. VLA-4 blockade by natalizumab inhibits sickle reticulocyte and leucocyte adhesion during simulated blood flow. Br J Haematol. 2016 Sep;174(6):970-82. doi: 10.1111/bjh.14158. Epub 2016 Jun 12. PMID: 27291690.
Very Late Antigen-4 (VLA-4, α4β1-integrin, ITGA4) orchestrates cell-cell and cell-endothelium adhesion. Given the proposed role of VLA-4 in sickle cell disease (SCD) pathophysiology, we evaluated the ability of the VLA-4 blocking antibody natalizumab to inhibit SCD blood cell adhesion. Natalizumab recognized surface VLA-4 on leucocytes and reticulocytes in whole blood from SCD subjects. SCD reticulocytes were positive for VLA-4, while VLA-4 staining of non-SCD reticulocytes was undetectable. Titrations with natalizumab revealed the presence of saturable levels of VLA-4 on both SCD reticulocytes and leucocytes similar to healthy subject leucocytes.
Alfano KM, Tarasev M, Meines S, Parunak G. An approach to measuring RBC haemolysis and profiling RBC mechanical fragility. J Med Eng Technol. 2016;40(4):162-71. doi: 10.3109/03091902.2016.1153741. Epub 2016 Mar 23. PMID: 27004768.
Red blood cells (RBC) can be damaged by medical products, from storage or from disease. Haemolysis (cell rupture and haemoglobin release) is often a key indicator, with mechanical fragility (MF) offering the potential to assess sub-haemolytic damage as well. This article reports on a unique approach to measuring haemolysis, without the need for centrifugation or other sample separation. It also reports on employing that in measuring blood fragility (susceptibility to haemolysis) under shear stress, utilising an electromagnet to cause a bead to oscillate within a cartridge that contains the sample.
White J, Lancelot M, Sarnaik S, Hines P. Increased erythrocyte adhesion to VCAM-1 during pulsatile flow: Application of a microfluidic flow adhesion bioassay. Clin Hemorheol Microcirc. 2015;60(2):201-13. doi: 10.3233/CH-141847. PMID: 24898561; PMCID: PMC4923762.
Sickle cell disease (SCD) is characterized by microvascular occlusion mediated by adhesive interactions of sickle erythrocytes (SSRBCs) to the endothelium. Most in vitro flow adhesion assays measure SSRBC adhesion during continuous flow, although in vivo SSRBC adhesive interactions occur during pulsatile flow. Using a well-plate microfluidic flow adhesion system, we demonstrate that isolated SSRBCs adhere to vascular cell adhesion molecule (VCAM-1) at greater levels during pulsatile versus continuous flow.
Tarasev M, Chakraborty S, Alfano K. RBC mechanical fragility as a direct blood quality metric to supplement storage time. Mil Med. 2015 Mar;180(3 Suppl):150-7. doi: 10.7205/MILMED-D-14-00404. PMID: 25747646.
Lengthy storage times and associated storage lesion can result in reduced red blood cell (RBC) efficacy, particularly dangerous for massively transfused patients. Today's inventory management makes storage times the de-factometric of blood quality. However, RBC units' quality may vary because of time-independent factors. Mechanical fragility (MF) of RBC, reflecting sub-lethal cell damage, can potentially provide a more physiologically relevant predictor of cell's performance “in vivo.”
Tarasev M, Alfano K, Chakraborty S, Light L, Doeden K, Gorlin JB. Similar donors-similar blood? Transfusion. 2014 Mar;54(3 Pt 2):933-41. doi: 10.1111/trf.12457. Epub 2013 Oct 28. PMID: 24660765.
Red blood cell (RBC) storage lesions have been suggested as contributing factors to suboptimal clinical outcomes. While undesirable effects of storage are well documented, their clinical relevance is still debated. Focus on storage time as the sole determinant of RBC quality ignores the variability in cell properties that may depend on factors other than age. Mechanical fragility (MF) aggregately reflects many storage‐related functional and structural changes. This study evaluates interdonor versus intradonor variability, throughout storage, of both MF and autohemolysis (AH).
Tarasev M, Alfano K, Chakraborty S, Bertholf M, Zubair A (2013)
Mechanical Fragility as a Potential Time-Independent Measure of Membrane
Integrity among Stored RBC Units. J Blood Disorders Transf 4:139.
Previous studies have shown that storage causes RBC membrane damage and subsequent potassium leakage to extracellular environment, with the effects exacerbated by RBC irradiation. While damage to RBC appears to worsen with storage time (ST), ST alone has not been shown to fully account for this phenomenon. It is therefore important to study the extent to which other time-independent factors can affect RBC membrane integrity. RBC mechanical fragility (MF) is evaluated as a surrogate measure of RBC membrane integrity due to its potential to reflect aggregate biochemical and biomechanical changes associated with storage.
Alfano K, Tarasev M. Investigating Direct Non-Age Metrics of Stored Blood Quality Loss. The Internet Journal of Medical
Technology. 2011 Volume 5 Number 1.
Long storage times for blood products are often unavoidable. Product age is essentially the only indicator used today for Red Blood Cell (RBC) quality loss during storage. Much controversy persists over the impact of RBC age on transfusion outcomes, as studies on this remain inconclusive. Such inconsistency may arise from unit-to-unit variability, which likely introduces some age-independence to RBC state. Thus, quality metrics other than storage time could aid with inventory management and/or treatment decisions. RBC membrane mechanical fragility is proposed here as one such candidate in vitro metric: it aggregately reflects a range of biochemical and biomechanical changes associated with storage lesion, and can provide a more comprehensive characterization of particular units than other properties.
Hines PC, McKnight TP, Seto W, Kwiatkowski JL. Central nervous system events in children with sickle cell disease presenting acutely with headache. J Pediatr. 2011 Sep;159(3):472-8. doi: 10.1016/j.jpeds.2011.02.009. Epub 2011 Mar 25. PMID: 21439575.
This is a retrospective cohort study of acute care visits for headache in children with SCD-SS. The prevalence of headache visits, neuroimaging evaluation, and acute CNS events were calculated and clinical and laboratory variables assessed.
Hines PC, Zen Q, Burney SN, Shea DA, Ataga KI, Orringer EP, Telen MJ, Parise LV. Novel epinephrine and cyclic AMP-mediated activation of BCAM/Lu-dependent sickle (SS) RBC adhesion. Blood. 2003 Apr 15;101(8):3281-7. doi: 10.1182/blood-2001-12-0289. Epub 2002 Dec 27. PMID: 12506027.
Sickle (SS) red blood cells (RBCs), unlike unaffected (AA) RBCs, adhere avidly to components of the vascular wall, and this abnormal adhesion is believed to contribute to the painful vaso- occlusive crises that occur in patients with sickle cell anemia. Laminin is an extracellular matrix (ECM) protein distributed throughout most vascular beds,1 with isoforms 10 and 11 specifi- cally supporting more SS RBC adhesion than other ECM proteins tested under physiologic shear and flow conditions.2 Patients with sickle cell anemia have extensive endothelial damage,3 likely bringing the underlying matrix laminin into direct contact with flowing blood.
Lee SP, Cunningham ML, Hines PC, Joneckis CC, Orringer EP, Parise LV. Sickle cell adhesion to laminin: potential role for the alpha5 chain. Blood. 1998 Oct 15;92(8):2951-8. PMID: 9763582.
Sickle red blood cell (RBC) adhesion to the endothelium and to exposed, underlying subendothelial proteins is believed to contribute to vascular occlusion in sickle cell disease. Laminin, a major component of the subendothelium, supports significant adhesion of sickle, but not normal RBCs. The purpose of this study was to define the adhesive region for sickle RBCs within a human laminin preparation using a flow adhesion assay designed to mimic physiologic flow through postcapillary venules.