The Fluid Membrane determines Mechanics of Red Blood Cell Extracellular Vesicles and is Softened in Hereditary Spherocytosis

AbstractExtracellular vesicles (EVs) are widely studied regarding their role in cell-to-cell communication and disease, as well as for applications as biomarker or drug delivery vehicle. EVs contain both membrane and intraluminal proteins, affecting their structural properties and thereby likely their functioning. Here, we use atomic force microscopy for the mechanical characterization of red blood cell (RBC) EVs from healthy individuals as well as from a patient with hereditary spherocytosis (HS) due to ankyrin deficiency. We show that the EVs are packed with proteins, yet their response to indentation is similar to that of a fluid lipid vesicle lacking proteins. The bending modulus of RBC EVs of healthy donors is ~15kbT, agreeing well with the bending modulus of the RBC membrane. Surprisingly, whereas RBCs become more rigid in HS, the excreted vesicles of a patient with this blood disorder have a significantly (~50%) lower bending modulus than donor EVs. These results shed new light on the mechanism and effects of EV budding and may underlie the reported increase in vesiculation and stiffening of RBCs in hereditary spherocytosis patients.

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Ektacytometry Analysis of Post-splenectomy Red Blood Cell Properties Identifies Cell Membrane Stability Test as a Novel Biomarker of Membrane Health in Hereditary Spherocytosis

Frontiers in Physiology ◽

10.3389/fphys.2021.641384 ◽

2021 ◽

Vol 12 ◽

Author(s):

M. C. Berrevoets ◽

J. Bos ◽

R. Huisjes ◽

T. H. Merkx ◽

B. A. van Oirschot ◽

...

Keyword(s):

Cell Membrane ◽

Blood Cell ◽

Red Blood Cell ◽

Clinical Benefit ◽

Hereditary Spherocytosis ◽

Case Control ◽

Stability Test ◽

Control Cohort ◽

Cell Membrane Stability ◽

Rbc Membrane

Hereditary spherocytosis (HS) is the most common form of hereditary chronic hemolytic anemia. It is caused by mutations in red blood cell (RBC) membrane and cytoskeletal proteins, which compromise membrane integrity, leading to vesiculation. Eventually, this leads to entrapment of poorly deformable spherocytes in the spleen. Splenectomy is a procedure often performed in HS. The clinical benefit results from removing the primary site of destruction, thereby improving RBC survival. But whether changes in RBC properties contribute to the clinical benefit of splenectomy is unknown. In this study we used ektacytometry to investigate the longitudinal effects of splenectomy on RBC properties in five well-characterized HS patients at four different time points and in a case-control cohort of 26 HS patients. Osmotic gradient ektacytometry showed that splenectomy resulted in improved intracellular viscosity (hydration state) whereas total surface area and surface-to-volume ratio remained essentially unchanged. The cell membrane stability test (CMST), which assesses the in vitro response to shear stress, showed that after splenectomy, HS RBCs had partly regained the ability to shed membrane, a property of healthy RBCs, which was confirmed in the case-control cohort. In particular the CMST holds promise as a novel biomarker in HS that reflects RBC membrane health and may be used to asses treatment response in HS.

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Exploratory spotlights on the proteome of extracellular vesicles in red blood cell units prepared by the same donors revealed that leukoreduction may matter for the quality and safety of transfusion therapy: Back to basics for research topics that might begin tomorrow’s realities

Transfusion and Apheresis Science ◽

10.1016/j.transci.2021.103165 ◽

2021 ◽

pp. 103165

Author(s):

Jerard Seghatchian

Keyword(s):

Blood Cell ◽

Extracellular Vesicles ◽

Red Blood Cell ◽

Quality And Safety ◽

Research Topics ◽

Transfusion Therapy

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Flow Cytometric Analysis of Erythrocytes Osmotic Fragility in Hereditary Spherocytosis: A Case‐Controlled Study Evaluating the Best Anticoagulant, Sample Pre‐Treatment and NaCl Concentration for Reliable Screening of this Red Blood Cell Membrane Disorder

Cytometry Part B Clinical Cytometry ◽

10.1002/cyto.b.21733 ◽

2018 ◽

Vol 94 (6) ◽

pp. 910-917

Author(s):

Camila Santos Nobre ◽

Jeovania Almeida Silva ◽

Rafael Henriques Jácomo ◽

Lídia Freire Abdalla Nery ◽

Gustavo Barcelos Barra

Keyword(s):

Cell Membrane ◽

Blood Cell ◽

Red Blood Cell ◽

Hereditary Spherocytosis ◽

Flow Cytometric Analysis ◽

Osmotic Fragility ◽

Controlled Study ◽

Flow Cytometric ◽

Nacl Concentration ◽

Pre Treatment

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Combined spectrin and ankyrin deficiency is common in autosomal dominant hereditary spherocytosis

Blood ◽

10.1182/blood.v82.10.2953.2953 ◽

1993 ◽

Vol 82 (10) ◽

pp. 2953-2960 ◽

Cited By ~ 47

Author(s):

P Savvides ◽

O Shalev ◽

KM John ◽

SE Lux

Keyword(s):

Blood Cell ◽

Red Blood Cell ◽

Autosomal Dominant ◽

Hereditary Spherocytosis ◽

Band 3 ◽

Normal Range ◽

Dominant Form ◽

Autosomal Dominant Form ◽

The Common ◽

Ankyrin Deficiency

Abstract The common autosomal dominant form of hereditary spherocytosis (HS) has been genetically linked to defects of the erythroid ankyrin gene in a few families; however, the frequency of ankyrin deficiency and its relationship to red blood cell (RBC) spectrin content are unknown. To test these questions, we measured RBC spectrin and ankyrin by radioimmunoassay in 39 patients from 20 families with dominant HS. Normal RBCs contained 242,000 +/- 20,500 spectrin heterodimers and 124,500 +/- 11,000 ankyrins per cell. In dominant HS, RBC spectrin and ankyrin ranged from about 40% to 100% of normal and were continuously distributed. Measurements in the same patient on different occasions were reproducible (+/- 5% to 10%) and RBCs from affected members of a kindred contained similar amounts of spectrin and ankyrin (+/- 3% to 4%). Spectrin and ankyrin levels were almost always less than the assay controls, but were less than the normal range in only 75% and 80% of kindreds, respectively. Remarkably, the degree of RBC spectrin and ankyrin deficiency was very similar in 19 of 20 HS kindreds. One otherwise typical family differed, with marked ankyrin deficiency (45% of control) and a relatively mild spectrin deficit (81%). We conclude that most patients with dominant HS have combined ankyrin and spectrin deficiency and that the two proteins are usually about equally deficient, suggesting that defects in ankyrin expression, ankyrin stability, or ankyrin band 3 (AE1) interactions may be common in dominant HS.

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Altered Levels of Desaturation and ω-6 Fatty Acids in Breast Cancer Patients’ Red Blood Cell Membranes

Metabolites ◽

10.3390/metabo10110469 ◽

2020 ◽

Vol 10 (11) ◽

pp. 469

Author(s):

Javier Amézaga ◽

Gurutze Ugartemendia ◽

Aitziber Larraioz ◽

Nerea Bretaña ◽

Aizpea Iruretagoyena ◽

...

Keyword(s):

Breast Cancer ◽

Fatty Acids ◽

Fatty Acid ◽

Blood Cell ◽

Cancer Patients ◽

Red Blood Cell ◽

Breast Cancer Patients ◽

Disease Evolution ◽

Lipidomic Analysis ◽

Rbc Membrane

Red blood cell (RBC) membrane can reflect fatty acid (FA) contribution from diet and biosynthesis. In cancer, membrane FAs are involved in tumorigenesis and invasiveness, and are indicated as biomarkers to monitor the disease evolution as well as potential targets for therapies and nutritional strategies. The present study provides RBC membrane FA profiles in recently diagnosed breast cancer patients before starting chemotherapy treatment. Patients and controls were recruited, and their dietary habits were collected. FA lipidomic analysis of mature erythrocyte membrane phospholipids in blood samples was performed. Data were adjusted to correct for the effects of diet, body mass index (BMI), and age, revealing that patients showed lower levels of saturated fatty acids (SFA) and higher levels of monounsaturated fatty acid, cis-vaccenic (25%) than controls, with consequent differences in desaturase enzymatic index (∆9 desaturase, –13.1%). In the case of polyunsaturated fatty acids (PUFA), patients had higher values of ω-6 FA (C18:2 (+11.1%); C20:4 (+7.4%)). RBC membrane lipidomic analysis in breast cancer revealed that ω-6 pathways are favored. These results suggest new potential targets for treatments and better nutritional guidelines.

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Red blood cell aquaporin-1 expression is decreased in hereditary spherocytosis

Annals of Hematology ◽

10.1007/s00277-016-2757-0 ◽

2016 ◽

Vol 95 (10) ◽

pp. 1595-1601 ◽

Cited By ~ 7

Author(s):

Renée L. Crisp ◽

Romina E. Maltaneri ◽

Daniela C. Vittori ◽

Liliana Solari ◽

Daniel Gammella ◽

...

Keyword(s):

Blood Cell ◽

Red Blood Cell ◽

Hereditary Spherocytosis ◽

Aquaporin 1

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Red blood cell (RBC) age at collection and storage influences RBC membrane-associated carbohydrates and lectin binding

Transfusion ◽

10.1111/j.1537-2995.2007.01230.x ◽

2007 ◽

Vol 47 (6) ◽

pp. 966-968 ◽

Cited By ~ 22

Author(s):

Rosemary L. Sparrow ◽

Margaret F. Veale ◽

Geraldine Healey ◽

Katherine A. Payne

Keyword(s):

Blood Cell ◽

Red Blood Cell ◽

Lectin Binding ◽

Rbc Membrane ◽

And Storage

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Plasmodium falciparum erythrocyte-binding antigen 175 triggers a biophysical change in the red blood cell that facilitates invasion

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1620843114 ◽

2017 ◽

Vol 114 (16) ◽

pp. 4225-4230 ◽

Cited By ~ 34

Author(s):

Marion Koch ◽

Katherine E. Wright ◽

Oliver Otto ◽

Maik Herbig ◽

Nichole D. Salinas ◽

...

Keyword(s):

Cell Surface ◽

Blood Cell ◽

Signaling Pathways ◽

Red Blood Cell ◽

Malaria Parasite ◽

Red Cell ◽

Biophysical Properties ◽

Parasite Invasion ◽

Bending Modulus ◽

Erythrocyte Binding

Invasion of the red blood cell (RBC) by the Plasmodium parasite defines the start of malaria disease pathogenesis. To date, experimental investigations into invasion have focused predominantly on the role of parasite adhesins or signaling pathways and the identity of binding receptors on the red cell surface. A potential role for signaling pathways within the erythrocyte, which might alter red cell biophysical properties to facilitate invasion, has largely been ignored. The parasite erythrocyte-binding antigen 175 (EBA175), a protein required for entry in most parasite strains, plays a key role by binding to glycophorin A (GPA) on the red cell surface, although the function of this binding interaction is unknown. Here, using real-time deformability cytometry and flicker spectroscopy to define biophysical properties of the erythrocyte, we show that EBA175 binding to GPA leads to an increase in the cytoskeletal tension of the red cell and a reduction in the bending modulus of the cell’s membrane. We isolate the changes in the cytoskeleton and membrane and show that reduction in the bending modulus is directly correlated with parasite invasion efficiency. These data strongly imply that the malaria parasite primes the erythrocyte surface through its binding antigens, altering the biophysical nature of the target cell and thus reducing a critical energy barrier to invasion. This finding would constitute a major change in our concept of malaria parasite invasion, suggesting it is, in fact, a balance between parasite and host cell physical forces working together to facilitate entry.

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