Biocompatible coupling of therapeutic fusion proteins to human erythrocytes

Villa, Carlos H.; Pan, Dongli; Johnston, Ian; Greineder, Colin F.; Walsh, Landis R.; Hood, Elizabeth D.; Cines, Douglas B.; Poncz, Mortimer; Siegel, Don L.; Muzykantov, Vladimir R.

doi:10.1182/bloodadvances.2017011734

Cited by 43 publications

(39 citation statements)

References 64 publications

(77 reference statements)

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Figure S5, Supporting Information, shows the result of a lysis assay where hybrid liposomes containing 20% DMPC were exposed to increasing osmotic stress by altering the molar concentration of a phosphate buffer saline. This method has previously been described as in vitro test for biocompatibiliy of RBC membranes 41. The data show an increase of lysed hybrid liposomes below 5.6 mM phosphate buffer saline, corresponding to a ≈150 mOsm solution, in good agreement with results for pure RBC 41.…”

Section: Discussionsupporting

confidence: 74%

“…This method has previously been described as in vitro test for biocompatibiliy of RBC membranes 41. The data show an increase of lysed hybrid liposomes below 5.6 mM phosphate buffer saline, corresponding to a ≈150 mOsm solution, in good agreement with results for pure RBC 41. It can, therefore, be expected that the hybrid liposomes will behave similar to RBCs in vivo.…”

Section: Discussionsupporting

confidence: 72%

See 1 more Smart Citation

Hybrid Erythrocyte Liposomes: Functionalized Red Blood Cell Membranes for Molecule Encapsulation

et al. 2020

View full text Add to dashboard Cite

The modification of erythrocyte membrane properties provides a new tool towards improved drug delivery and biomedical applications. The fabrication of hybrid erythrocyte liposomes is presented by doping red blood cell membranes with synthetic lipid molecules of different classes (PC, PS, PG) and different degrees of saturation (14:0, 16:0–18:1). The respective solubility limits are determined, and material properties of the hybrid liposomes are studied by a combination of X‐ray diffraction, epi‐fluorescent microscopy, dynamic light scattering (DLS), Zeta potential, UV‐vis spectroscopy, and Molecular Dynamics (MD) simulations. Membrane thickness and lipid orientation can be tuned through the addition of phosphatidylcholine lipids. The hybrid membranes can be fluorescently labelled by incorporating Texas‐red DHPE, and their charge modified by incorporating phosphatidylserine and phosphatidylglycerol. By using fluorescein labeled dextran as an example, it is demonstrated that small molecules can be encapsulated into these hybrid liposomes.

show abstract

Section: Discussionsupporting

confidence: 74%

Section: Discussionsupporting

confidence: 72%

Hybrid Erythrocyte Liposomes: Functionalized Red Blood Cell Membranes for Molecule Encapsulation

et al. 2020

View full text Add to dashboard Cite

show abstract

“…Differences in effects on RBCs by coupling approaches can be further exacerbated when comparing between different epitopes. For example, our group recently showed that scFv-thrombomodulin fusion proteins directed against human RhCE did not impact RBC rigidity and fragility, while those targeted to glycophorin A significantly increased these markers of RBC damage [114]. This suggests that features of the molecule used to couple to the surface of the RBC (e.g., avidity, Fc fragment, epitope) have a significant impact on the biocompatibility of the loading strategy.…”

Section: Surface Loading Of Rbc-impact Of Dose and Coupling Strategymentioning

confidence: 99%

Vascular Drug Delivery Using Carrier Red Blood Cells: Focus on RBC Surface Loading and Pharmacokinetics

et al. 2020

Self Cite

View full text Add to dashboard Cite

Red blood cells (RBC) have great potential as drug delivery systems, capable of producing unprecedented changes in pharmacokinetics, pharmacodynamics, and immunogenicity. Despite this great potential and nearly 50 years of research, it is only recently that RBC-mediated drug delivery has begun to move out of the academic lab and into industrial drug development. RBC loading with drugs can be performed in several ways-either via encapsulation within the RBC or surface coupling, and either ex vivo or in vivo-depending on the intended application. In this review, we briefly summarize currently used technologies for RBC loading/coupling with an eye on how pharmacokinetics is impacted. Additionally, we provide a detailed description of key ADME (absorption, distribution, metabolism, elimination) changes that would be expected for RBC-associated drugs and address unique features of RBC pharmacokinetics. As thorough understanding of pharmacokinetics is critical in successful translation to the clinic, we expect that this review will provide a jumping off point for further investigations into this area.Pharmaceutics 2020, 12, 440 2 of 21 of the research enterprise encompassing the design of drug delivery systems (DDS)-liposomes, antibody-drug conjugates, and polymeric nanocarriers, to name a few.Nevertheless, approaches to use RBCs as carriers for pharmacological agents have recently gained significant and rapidly growing attention. Progress in this field is rapidly diversifying and accelerating towards potentially clinically useful products. Many groups are now investigating the use of RBCs in drug delivery and are making significant contributions, leading to breakthrough findings and upbeat investments. Several RBC-based drug delivery approaches have entered clinical trials, including RBC-encapsulated asparaginase (Erytech, Phase 3) and dexamethasone (EryDel, Phase 3). Novel advanced strategies are emerging, including genetic molecular modifications of RBC [2,3], modulation of the immune system by RBC-coupled antigens [3,4], and vascular transfer of RBC-coupled nanocarriers (RBC hitchhiking) [5][6][7].Both encapsulation into and coupling to the surface of RBC fundamentally transform the key parameters of absorption, distribution, metabolism, and elimination (ADME) of drugs and drug delivery systems (DDS), including diverse nanocarriers. To our knowledge, studies of the pharmacokinetics (PK) and pharmacodynamics (PD) of RBC-based DDS are lacking, despite great relevance for industrial development and clinical utility.In order to help to close this gap of knowledge, in this paper we undertook the first attempt to define specific, salient parameters controlling behavior of RBC/DDS in the body. Our goal is to provide the modular framework for experimental and theoretical pre-clinical and clinical investigations of ADME-PK-PD features of RBC-based drug delivery.

show abstract

“…Pro [18][19][20][21][22][23][24][25] RBCs release membrane-derived procoagulant microvesicles bearing phosphatidylserine during in vivo aging and in vitro storage Pro [28][29][30][31]33,34] Meizothrombin, a protein C activator with low fibrinogen-cleaving activity, is formed on RBCs and released into the blood Anti [20] Factor IX is activated directly by an elastase-like enzyme on the RBC Pro [31,32,34,36,37,[40][41][42][43][44][45][47][48][49] Quantitative and qualitative changes in RBCs related to bleeding and thrombosis…”

Section: Introductionmentioning

confidence: 99%

Red blood cells: the forgotten player in hemostasis and thrombosis

Weisel

Litvinov

2019

Journal of Thrombosis and Haemostasis

294

272

View full text Add to dashboard Cite

Summary New evidence has stirred up a long‐standing but undeservedly forgotten interest in the role of erythrocytes, or red blood cells (RBCs), in blood clotting and its disorders. This review summarizes the most recent research that describes the involvement of RBCs in hemostasis and thrombosis. There are both quantitative and qualitative changes in RBCs that affect bleeding and thrombosis, as well as interactions of RBCs with cellular and molecular components of the hemostatic system. The changes in RBCs that affect hemostasis and thrombosis include RBC counts or hematocrit (modulating blood rheology through viscosity) and qualitative changes, such as deformability, aggregation, expression of adhesive proteins and phosphatidylserine, release of extracellular microvesicles, and hemolysis. The pathogenic mechanisms implicated in thrombotic and hemorrhagic risk include variable adherence of RBCs to the vessel wall, which depends on the functional state of RBCs and/or endothelium, modulation of platelet reactivity and platelet margination, alterations of fibrin structure and reduced susceptibility to fibrinolysis, modulation of nitric oxide availability, and the levels of von Willebrand factor and factor VIII in blood related to the ABO blood group system. RBCs are involved in platelet‐driven contraction of clots and thrombi that results in formation of a tightly packed array of polyhedral erythrocytes, or polyhedrocytes, which comprises a nearly impermeable barrier that is important for hemostasis and wound healing. The revisited notion of the importance of RBCs is largely based on clinical and experimental associations between RBCs and thrombosis or bleeding, implying that RBCs are a prospective therapeutic target in hemostatic and thrombotic disorders.

show abstract

Biocompatible coupling of therapeutic fusion proteins to human erythrocytes

Cited by 43 publications

References 64 publications

Hybrid Erythrocyte Liposomes: Functionalized Red Blood Cell Membranes for Molecule Encapsulation

Hybrid Erythrocyte Liposomes: Functionalized Red Blood Cell Membranes for Molecule Encapsulation

Vascular Drug Delivery Using Carrier Red Blood Cells: Focus on RBC Surface Loading and Pharmacokinetics

Red blood cells: the forgotten player in hemostasis and thrombosis

Contact Info

Product

Resources

About