Genetic transfer of fusion proteins effectively inhibits VCAM-1-mediated cell adhesion and transmigration via inhibition of cytoskeletal anchorage

Christoph Eugen Hagemeyer, Ingo Ahrens, Nicole Bassler, Natia Dschachutaschwili, Yung-Chih Chen, Steffen U Eisenhardt, Christoph Bode, Karlheinz Peter

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

Abstract

The adhesion of leukocytes to endothelium plays a central role in the development of atherosclerosis and thus represents an attractive therapeutic target for anti-atherosclerotic therapies. Vascular cell adhesion molecule-1 (VCAM-1) mediates both the initial tethering and the firm adhesion of leukocytes to endothelial cells. Our work evaluates the feasibility of using the cytoskeletal anchorage of VCAM-1 as a target for gene therapy. As a proof of concept, integrin aIIb?3-mediated cell adhesion with clearly defined cytoskeletal anchorage was tested. We constructed fusion proteins containing the intracellular domain of ?3 placed at various distances to the cell membrane. Using cell adhesion assays and immunofluorescence, we established fusion constructs with competitive and dominant negative inhibition of cell adhesion. With the goal being the transfer of the dominant negative mechanism towards VCAM-1 inhibition, we constructed a fusion molecule containing the cytoplasmic domain of VCAM-1. Indeed, VCAM-1 mediated leukocyte adhesion can be inhibited via transfection of DNA encoding the designed VCAM-1 fusion protein. This is demonstrated in adhesion assays under static and flow conditions using CHO cells expressing recombinant VCAM-1 as well as activated endothelial cells. Thus, we are able to describe a novel approach for dominant negative inhibition of leukocyte adhesion to endothelial cells. This approach warrants further development as a novel gene therapeutic strategy that aims for a locally restricted effect at atherosclerotic areas of the vasculature.
Original languageEnglish
Pages (from-to)290 - 302
Number of pages13
JournalJournal of Cellular and Molecular Medicine
Volume14
Issue number1-2
DOIs
Publication statusPublished - 2010
Externally publishedYes

Cite this

Hagemeyer, Christoph Eugen ; Ahrens, Ingo ; Bassler, Nicole ; Dschachutaschwili, Natia ; Chen, Yung-Chih ; Eisenhardt, Steffen U ; Bode, Christoph ; Peter, Karlheinz. / Genetic transfer of fusion proteins effectively inhibits VCAM-1-mediated cell adhesion and transmigration via inhibition of cytoskeletal anchorage. In: Journal of Cellular and Molecular Medicine. 2010 ; Vol. 14, No. 1-2. pp. 290 - 302.
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title = "Genetic transfer of fusion proteins effectively inhibits VCAM-1-mediated cell adhesion and transmigration via inhibition of cytoskeletal anchorage",
abstract = "The adhesion of leukocytes to endothelium plays a central role in the development of atherosclerosis and thus represents an attractive therapeutic target for anti-atherosclerotic therapies. Vascular cell adhesion molecule-1 (VCAM-1) mediates both the initial tethering and the firm adhesion of leukocytes to endothelial cells. Our work evaluates the feasibility of using the cytoskeletal anchorage of VCAM-1 as a target for gene therapy. As a proof of concept, integrin aIIb?3-mediated cell adhesion with clearly defined cytoskeletal anchorage was tested. We constructed fusion proteins containing the intracellular domain of ?3 placed at various distances to the cell membrane. Using cell adhesion assays and immunofluorescence, we established fusion constructs with competitive and dominant negative inhibition of cell adhesion. With the goal being the transfer of the dominant negative mechanism towards VCAM-1 inhibition, we constructed a fusion molecule containing the cytoplasmic domain of VCAM-1. Indeed, VCAM-1 mediated leukocyte adhesion can be inhibited via transfection of DNA encoding the designed VCAM-1 fusion protein. This is demonstrated in adhesion assays under static and flow conditions using CHO cells expressing recombinant VCAM-1 as well as activated endothelial cells. Thus, we are able to describe a novel approach for dominant negative inhibition of leukocyte adhesion to endothelial cells. This approach warrants further development as a novel gene therapeutic strategy that aims for a locally restricted effect at atherosclerotic areas of the vasculature.",
author = "Hagemeyer, {Christoph Eugen} and Ingo Ahrens and Nicole Bassler and Natia Dschachutaschwili and Yung-Chih Chen and Eisenhardt, {Steffen U} and Christoph Bode and Karlheinz Peter",
year = "2010",
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Hagemeyer, CE, Ahrens, I, Bassler, N, Dschachutaschwili, N, Chen, Y-C, Eisenhardt, SU, Bode, C & Peter, K 2010, 'Genetic transfer of fusion proteins effectively inhibits VCAM-1-mediated cell adhesion and transmigration via inhibition of cytoskeletal anchorage', Journal of Cellular and Molecular Medicine, vol. 14, no. 1-2, pp. 290 - 302. https://doi.org/10.1111/j.1582-4934.2008.00409.x

Genetic transfer of fusion proteins effectively inhibits VCAM-1-mediated cell adhesion and transmigration via inhibition of cytoskeletal anchorage. / Hagemeyer, Christoph Eugen; Ahrens, Ingo; Bassler, Nicole; Dschachutaschwili, Natia; Chen, Yung-Chih; Eisenhardt, Steffen U; Bode, Christoph; Peter, Karlheinz.

In: Journal of Cellular and Molecular Medicine, Vol. 14, No. 1-2, 2010, p. 290 - 302.

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

TY - JOUR

T1 - Genetic transfer of fusion proteins effectively inhibits VCAM-1-mediated cell adhesion and transmigration via inhibition of cytoskeletal anchorage

AU - Hagemeyer, Christoph Eugen

AU - Ahrens, Ingo

AU - Bassler, Nicole

AU - Dschachutaschwili, Natia

AU - Chen, Yung-Chih

AU - Eisenhardt, Steffen U

AU - Bode, Christoph

AU - Peter, Karlheinz

PY - 2010

Y1 - 2010

N2 - The adhesion of leukocytes to endothelium plays a central role in the development of atherosclerosis and thus represents an attractive therapeutic target for anti-atherosclerotic therapies. Vascular cell adhesion molecule-1 (VCAM-1) mediates both the initial tethering and the firm adhesion of leukocytes to endothelial cells. Our work evaluates the feasibility of using the cytoskeletal anchorage of VCAM-1 as a target for gene therapy. As a proof of concept, integrin aIIb?3-mediated cell adhesion with clearly defined cytoskeletal anchorage was tested. We constructed fusion proteins containing the intracellular domain of ?3 placed at various distances to the cell membrane. Using cell adhesion assays and immunofluorescence, we established fusion constructs with competitive and dominant negative inhibition of cell adhesion. With the goal being the transfer of the dominant negative mechanism towards VCAM-1 inhibition, we constructed a fusion molecule containing the cytoplasmic domain of VCAM-1. Indeed, VCAM-1 mediated leukocyte adhesion can be inhibited via transfection of DNA encoding the designed VCAM-1 fusion protein. This is demonstrated in adhesion assays under static and flow conditions using CHO cells expressing recombinant VCAM-1 as well as activated endothelial cells. Thus, we are able to describe a novel approach for dominant negative inhibition of leukocyte adhesion to endothelial cells. This approach warrants further development as a novel gene therapeutic strategy that aims for a locally restricted effect at atherosclerotic areas of the vasculature.

AB - The adhesion of leukocytes to endothelium plays a central role in the development of atherosclerosis and thus represents an attractive therapeutic target for anti-atherosclerotic therapies. Vascular cell adhesion molecule-1 (VCAM-1) mediates both the initial tethering and the firm adhesion of leukocytes to endothelial cells. Our work evaluates the feasibility of using the cytoskeletal anchorage of VCAM-1 as a target for gene therapy. As a proof of concept, integrin aIIb?3-mediated cell adhesion with clearly defined cytoskeletal anchorage was tested. We constructed fusion proteins containing the intracellular domain of ?3 placed at various distances to the cell membrane. Using cell adhesion assays and immunofluorescence, we established fusion constructs with competitive and dominant negative inhibition of cell adhesion. With the goal being the transfer of the dominant negative mechanism towards VCAM-1 inhibition, we constructed a fusion molecule containing the cytoplasmic domain of VCAM-1. Indeed, VCAM-1 mediated leukocyte adhesion can be inhibited via transfection of DNA encoding the designed VCAM-1 fusion protein. This is demonstrated in adhesion assays under static and flow conditions using CHO cells expressing recombinant VCAM-1 as well as activated endothelial cells. Thus, we are able to describe a novel approach for dominant negative inhibition of leukocyte adhesion to endothelial cells. This approach warrants further development as a novel gene therapeutic strategy that aims for a locally restricted effect at atherosclerotic areas of the vasculature.

UR - http://onlinelibrary.wiley.com/doi/10.1111/j.1582-4934.2008.00409.x/abstract;jsessionid=7B04BACEE7A87B2D449DBA3E8D6387CB.f04t02

U2 - 10.1111/j.1582-4934.2008.00409.x

DO - 10.1111/j.1582-4934.2008.00409.x

M3 - Article

VL - 14

SP - 290

EP - 302

JO - Journal of Cellular and Molecular Medicine

JF - Journal of Cellular and Molecular Medicine

SN - 1582-1838

IS - 1-2

ER -

Hagemeyer CE, Ahrens I, Bassler N, Dschachutaschwili N, Chen Y-C, Eisenhardt SU et al. Genetic transfer of fusion proteins effectively inhibits VCAM-1-mediated cell adhesion and transmigration via inhibition of cytoskeletal anchorage. Journal of Cellular and Molecular Medicine. 2010;14(1-2):290 - 302. https://doi.org/10.1111/j.1582-4934.2008.00409.x

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