Characterization of gpvi-or gpvi-cd39-coated nanoparticles and their impact on in vitro thrombus formation

Jeremy A. Nestele; Anne Katrin Rohlfing; Valerie Dicenta; Alexander Bild; Daniela Eißler; Frederic Emschermann; Marcel Kremser; Konstantin Krutzke; Tilman E. Schäffer; Oliver Borst; Moran Levi; Netanel Korin; Meinrad Paul Gawaz

doi:10.3390/ijms23010011

Characterization of gpvi-or gpvi-cd39-coated nanoparticles and their impact on in vitro thrombus formation

Jeremy A. Nestele, Anne Katrin Rohlfing, Valerie Dicenta, Alexander Bild, Daniela Eißler, Frederic Emschermann, Marcel Kremser, Konstantin Krutzke, Tilman E. Schäffer, Oliver Borst, Moran Levi, Netanel Korin, Meinrad Paul Gawaz

Technion - Israel Institute of Technology

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

Abstract

Traditional antithrombotic agents commonly share a therapy-limiting side effect, as they increase the overall systemic bleeding risk. A novel approach for targeted antithrombotic therapy is nanoparticles. In other therapeutic fields, nanoparticles have enabled site-specific delivery with low levels of toxicity and side effects. Here, we paired nanotechnology with an established dimeric glyco-protein VI-Fc (GPVI-Fc) and a GPVI-CD39 fusion protein, thereby combining site-specific delivery and new antithrombotic drugs. Poly(lactic-co-glycolic acid) (PLGA) nanoparticles, NP-BSA, NP-GPVI and NP-GPVI-CD39 were characterized through electron microscopy, atomic force measurements and flow cytometry. Light transmission aggregometry enabled analysis of platelet aggregation. Throm-bus formation was observed through flow chamber experiments. NP-GPVI and NP-GPVI-CD39 displayed a characteristic surface coating pattern. Fluorescence properties were identical amongst all samples. NP-GPVI and NP-GPVI-CD39 significantly impaired platelet aggregation. Thrombus formation was significantly impaired by NP-GPVI and was particularly impaired by NP-GPVI-CD39. The receptor-coated nanoparticles NP-GPVI and the bifunctional molecule NP-GPVI-CD39 demonstrated significant inhibition of in vitro thrombus formation. Consequently, the nanoparticle-mediated antithrombotic effect of GPVI-Fc, as well as GPVI-CD39, and an additive impact of CD39 was confirmed. In conclusion, NP-GPVI and NP-GPVI-CD39 may serve as a promising foundation for a novel therapeutic approach regarding targeted antithrombotic therapy.

Original language	English
Article number	11
Journal	INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
Volume	23
Issue number	1
DOIs	https://doi.org/10.3390/ijms23010011
State	Published - 21 Dec 2021

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

SDG 3 Good Health and Well-being

Keywords

Antigens, CD/metabolism
Apyrase/metabolism
Fibrinolytic Agents/pharmacology
Flow Cytometry
Humans
Immunoglobulin Fc Fragments/metabolism
In Vitro Techniques
Microscopy, Electron
Models, Biological
Nanoparticles
Particle Size
Platelet Aggregation/drug effects
Platelet Membrane Glycoproteins/metabolism
Polylactic Acid-Polyglycolic Acid Copolymer/chemistry
Recombinant Fusion Proteins/pharmacology

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10.3390/ijms23010011

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Nestele, J. A., Rohlfing, A. K., Dicenta, V., Bild, A., Eißler, D., Emschermann, F., Kremser, M., Krutzke, K., Schäffer, T. E., Borst, O., Levi, M., Korin, N., & Gawaz, M. P. (2021). Characterization of gpvi-or gpvi-cd39-coated nanoparticles and their impact on in vitro thrombus formation. INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES, 23(1), Article 11. https://doi.org/10.3390/ijms23010011

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title = "Characterization of gpvi-or gpvi-cd39-coated nanoparticles and their impact on in vitro thrombus formation",

abstract = "Traditional antithrombotic agents commonly share a therapy-limiting side effect, as they increase the overall systemic bleeding risk. A novel approach for targeted antithrombotic therapy is nanoparticles. In other therapeutic fields, nanoparticles have enabled site-specific delivery with low levels of toxicity and side effects. Here, we paired nanotechnology with an established dimeric glyco-protein VI-Fc (GPVI-Fc) and a GPVI-CD39 fusion protein, thereby combining site-specific delivery and new antithrombotic drugs. Poly(lactic-co-glycolic acid) (PLGA) nanoparticles, NP-BSA, NP-GPVI and NP-GPVI-CD39 were characterized through electron microscopy, atomic force measurements and flow cytometry. Light transmission aggregometry enabled analysis of platelet aggregation. Throm-bus formation was observed through flow chamber experiments. NP-GPVI and NP-GPVI-CD39 displayed a characteristic surface coating pattern. Fluorescence properties were identical amongst all samples. NP-GPVI and NP-GPVI-CD39 significantly impaired platelet aggregation. Thrombus formation was significantly impaired by NP-GPVI and was particularly impaired by NP-GPVI-CD39. The receptor-coated nanoparticles NP-GPVI and the bifunctional molecule NP-GPVI-CD39 demonstrated significant inhibition of in vitro thrombus formation. Consequently, the nanoparticle-mediated antithrombotic effect of GPVI-Fc, as well as GPVI-CD39, and an additive impact of CD39 was confirmed. In conclusion, NP-GPVI and NP-GPVI-CD39 may serve as a promising foundation for a novel therapeutic approach regarding targeted antithrombotic therapy.",

keywords = "Antigens, CD/metabolism, Apyrase/metabolism, Fibrinolytic Agents/pharmacology, Flow Cytometry, Humans, Immunoglobulin Fc Fragments/metabolism, In Vitro Techniques, Microscopy, Electron, Models, Biological, Nanoparticles, Particle Size, Platelet Aggregation/drug effects, Platelet Membrane Glycoproteins/metabolism, Polylactic Acid-Polyglycolic Acid Copolymer/chemistry, Recombinant Fusion Proteins/pharmacology",

author = "Nestele, \{Jeremy A.\} and Rohlfing, \{Anne Katrin\} and Valerie Dicenta and Alexander Bild and Daniela Ei{\ss}ler and Frederic Emschermann and Marcel Kremser and Konstantin Krutzke and Sch{\"a}ffer, \{Tilman E.\} and Oliver Borst and Moran Levi and Netanel Korin and Gawaz, \{Meinrad Paul\}",

note = "Publisher Copyright: {\textcopyright} 2021 by the authors. Licensee MDPI, Basel, Switzerland.",

year = "2021",

month = dec,

day = "21",

doi = "10.3390/ijms23010011",

language = "الإنجليزيّة",

volume = "23",

journal = "INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES",

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Nestele, JA, Rohlfing, AK, Dicenta, V, Bild, A, Eißler, D, Emschermann, F, Kremser, M, Krutzke, K, Schäffer, TE, Borst, O, Levi, M, Korin, N & Gawaz, MP 2021, 'Characterization of gpvi-or gpvi-cd39-coated nanoparticles and their impact on in vitro thrombus formation', INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES, vol. 23, no. 1, 11. https://doi.org/10.3390/ijms23010011

TY - JOUR

T1 - Characterization of gpvi-or gpvi-cd39-coated nanoparticles and their impact on in vitro thrombus formation

AU - Nestele, Jeremy A.

AU - Rohlfing, Anne Katrin

AU - Dicenta, Valerie

AU - Bild, Alexander

AU - Eißler, Daniela

AU - Emschermann, Frederic

AU - Kremser, Marcel

AU - Krutzke, Konstantin

AU - Schäffer, Tilman E.

AU - Borst, Oliver

AU - Levi, Moran

AU - Korin, Netanel

AU - Gawaz, Meinrad Paul

PY - 2021/12/21

Y1 - 2021/12/21

N2 - Traditional antithrombotic agents commonly share a therapy-limiting side effect, as they increase the overall systemic bleeding risk. A novel approach for targeted antithrombotic therapy is nanoparticles. In other therapeutic fields, nanoparticles have enabled site-specific delivery with low levels of toxicity and side effects. Here, we paired nanotechnology with an established dimeric glyco-protein VI-Fc (GPVI-Fc) and a GPVI-CD39 fusion protein, thereby combining site-specific delivery and new antithrombotic drugs. Poly(lactic-co-glycolic acid) (PLGA) nanoparticles, NP-BSA, NP-GPVI and NP-GPVI-CD39 were characterized through electron microscopy, atomic force measurements and flow cytometry. Light transmission aggregometry enabled analysis of platelet aggregation. Throm-bus formation was observed through flow chamber experiments. NP-GPVI and NP-GPVI-CD39 displayed a characteristic surface coating pattern. Fluorescence properties were identical amongst all samples. NP-GPVI and NP-GPVI-CD39 significantly impaired platelet aggregation. Thrombus formation was significantly impaired by NP-GPVI and was particularly impaired by NP-GPVI-CD39. The receptor-coated nanoparticles NP-GPVI and the bifunctional molecule NP-GPVI-CD39 demonstrated significant inhibition of in vitro thrombus formation. Consequently, the nanoparticle-mediated antithrombotic effect of GPVI-Fc, as well as GPVI-CD39, and an additive impact of CD39 was confirmed. In conclusion, NP-GPVI and NP-GPVI-CD39 may serve as a promising foundation for a novel therapeutic approach regarding targeted antithrombotic therapy.

AB - Traditional antithrombotic agents commonly share a therapy-limiting side effect, as they increase the overall systemic bleeding risk. A novel approach for targeted antithrombotic therapy is nanoparticles. In other therapeutic fields, nanoparticles have enabled site-specific delivery with low levels of toxicity and side effects. Here, we paired nanotechnology with an established dimeric glyco-protein VI-Fc (GPVI-Fc) and a GPVI-CD39 fusion protein, thereby combining site-specific delivery and new antithrombotic drugs. Poly(lactic-co-glycolic acid) (PLGA) nanoparticles, NP-BSA, NP-GPVI and NP-GPVI-CD39 were characterized through electron microscopy, atomic force measurements and flow cytometry. Light transmission aggregometry enabled analysis of platelet aggregation. Throm-bus formation was observed through flow chamber experiments. NP-GPVI and NP-GPVI-CD39 displayed a characteristic surface coating pattern. Fluorescence properties were identical amongst all samples. NP-GPVI and NP-GPVI-CD39 significantly impaired platelet aggregation. Thrombus formation was significantly impaired by NP-GPVI and was particularly impaired by NP-GPVI-CD39. The receptor-coated nanoparticles NP-GPVI and the bifunctional molecule NP-GPVI-CD39 demonstrated significant inhibition of in vitro thrombus formation. Consequently, the nanoparticle-mediated antithrombotic effect of GPVI-Fc, as well as GPVI-CD39, and an additive impact of CD39 was confirmed. In conclusion, NP-GPVI and NP-GPVI-CD39 may serve as a promising foundation for a novel therapeutic approach regarding targeted antithrombotic therapy.

KW - Antigens, CD/metabolism

KW - Apyrase/metabolism

KW - Fibrinolytic Agents/pharmacology

KW - Flow Cytometry

KW - Humans

KW - Immunoglobulin Fc Fragments/metabolism

KW - In Vitro Techniques

KW - Microscopy, Electron

KW - Models, Biological

KW - Nanoparticles

KW - Particle Size

KW - Platelet Aggregation/drug effects

KW - Platelet Membrane Glycoproteins/metabolism

KW - Polylactic Acid-Polyglycolic Acid Copolymer/chemistry

KW - Recombinant Fusion Proteins/pharmacology

UR - http://www.scopus.com/inward/record.url?scp=85121372279&partnerID=8YFLogxK

U2 - 10.3390/ijms23010011

DO - 10.3390/ijms23010011

M3 - مقالة

C2 - 35008437

SN - 1661-6596

VL - 23

JO - INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES

JF - INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES

IS - 1

M1 - 11

ER -

Characterization of gpvi-or gpvi-cd39-coated nanoparticles and their impact on in vitro thrombus formation

Abstract

UN SDGs

Keywords

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