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Imaging, myeloid precursor immortalization, and genome editing for defining mechanisms of leukocyte recruitment in vivo

Recruitment of leukocytes from the blood to sites of inflammation poses a promising target for new diagnostic and therapeutic approaches. We aimed to develop a novel method to non-invasively analyze molecular mechanisms of leukocyte migration in pre-clinical models of inflammation in vivo. Methods:...

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Autores principales: Gran, Sandra, Honold, Lisa, Fehler, Olesja, Zenker, Stefanie, Eligehausen, Sarah, Kuhlmann, Michael T., Geven, Edwin, van den Bosch, Martijn, van Lent, Peter, Spiekermann, Christoph, Hermann, Sven, Vogl, Thomas, Schäfers, Michael, Roth, Johannes
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Ivyspring International Publisher 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5928898/
https://www.ncbi.nlm.nih.gov/pubmed/29721088
http://dx.doi.org/10.7150/thno.23632
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author Gran, Sandra
Honold, Lisa
Fehler, Olesja
Zenker, Stefanie
Eligehausen, Sarah
Kuhlmann, Michael T.
Geven, Edwin
van den Bosch, Martijn
van Lent, Peter
Spiekermann, Christoph
Hermann, Sven
Vogl, Thomas
Schäfers, Michael
Roth, Johannes
author_facet Gran, Sandra
Honold, Lisa
Fehler, Olesja
Zenker, Stefanie
Eligehausen, Sarah
Kuhlmann, Michael T.
Geven, Edwin
van den Bosch, Martijn
van Lent, Peter
Spiekermann, Christoph
Hermann, Sven
Vogl, Thomas
Schäfers, Michael
Roth, Johannes
author_sort Gran, Sandra
collection PubMed
description Recruitment of leukocytes from the blood to sites of inflammation poses a promising target for new diagnostic and therapeutic approaches. We aimed to develop a novel method to non-invasively analyze molecular mechanisms of leukocyte migration in pre-clinical models of inflammation in vivo. Methods: We used the ER-HoxB8 system to transiently immortalize murine myeloid precursors from wildtype and CD18- as well as MRP14-deficient mice. A VLA4α-/- cell line was generated by CRISPR/Cas9-mediated gene editing. We analyzed the migration of wildtype and knockout leukocytes in vivo by optical and nuclear imaging in mice with irritant contact dermatitis, cutaneous granuloma, experimental arthritis and myocardial infarction. Results: Transient immortalization, gene editing and in vivo imaging can be combined to analyze migratory mechanisms of murine leukocytes, even for gene deletions resulting in lethal phenotypes in mice. We reliably confirmed known migratory defects of leukocytes deficient for the adhesion molecules CD18 or VLA4α. Also, using our new method we identified a new role of the most abundant calcium-binding proteins in phagocytes and major alarmins in many inflammatory diseases, MRP8 and MRP14, for transmigration in vivo. Conclusion: We provide a combinatorial approach to rapidly characterize molecular mechanisms of leukocyte recruitment in vivo, with the potential to aid in identification of diagnostic and therapeutic targets in inflammatory pathologies.
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spelling pubmed-59288982018-05-02 Imaging, myeloid precursor immortalization, and genome editing for defining mechanisms of leukocyte recruitment in vivo Gran, Sandra Honold, Lisa Fehler, Olesja Zenker, Stefanie Eligehausen, Sarah Kuhlmann, Michael T. Geven, Edwin van den Bosch, Martijn van Lent, Peter Spiekermann, Christoph Hermann, Sven Vogl, Thomas Schäfers, Michael Roth, Johannes Theranostics Research Paper Recruitment of leukocytes from the blood to sites of inflammation poses a promising target for new diagnostic and therapeutic approaches. We aimed to develop a novel method to non-invasively analyze molecular mechanisms of leukocyte migration in pre-clinical models of inflammation in vivo. Methods: We used the ER-HoxB8 system to transiently immortalize murine myeloid precursors from wildtype and CD18- as well as MRP14-deficient mice. A VLA4α-/- cell line was generated by CRISPR/Cas9-mediated gene editing. We analyzed the migration of wildtype and knockout leukocytes in vivo by optical and nuclear imaging in mice with irritant contact dermatitis, cutaneous granuloma, experimental arthritis and myocardial infarction. Results: Transient immortalization, gene editing and in vivo imaging can be combined to analyze migratory mechanisms of murine leukocytes, even for gene deletions resulting in lethal phenotypes in mice. We reliably confirmed known migratory defects of leukocytes deficient for the adhesion molecules CD18 or VLA4α. Also, using our new method we identified a new role of the most abundant calcium-binding proteins in phagocytes and major alarmins in many inflammatory diseases, MRP8 and MRP14, for transmigration in vivo. Conclusion: We provide a combinatorial approach to rapidly characterize molecular mechanisms of leukocyte recruitment in vivo, with the potential to aid in identification of diagnostic and therapeutic targets in inflammatory pathologies. Ivyspring International Publisher 2018-03-23 /pmc/articles/PMC5928898/ /pubmed/29721088 http://dx.doi.org/10.7150/thno.23632 Text en © Ivyspring International Publisher This is an open access article distributed under the terms of the Creative Commons Attribution (CC BY-NC) license (https://creativecommons.org/licenses/by-nc/4.0/). See http://ivyspring.com/terms for full terms and conditions.
spellingShingle Research Paper
Gran, Sandra
Honold, Lisa
Fehler, Olesja
Zenker, Stefanie
Eligehausen, Sarah
Kuhlmann, Michael T.
Geven, Edwin
van den Bosch, Martijn
van Lent, Peter
Spiekermann, Christoph
Hermann, Sven
Vogl, Thomas
Schäfers, Michael
Roth, Johannes
Imaging, myeloid precursor immortalization, and genome editing for defining mechanisms of leukocyte recruitment in vivo
title Imaging, myeloid precursor immortalization, and genome editing for defining mechanisms of leukocyte recruitment in vivo
title_full Imaging, myeloid precursor immortalization, and genome editing for defining mechanisms of leukocyte recruitment in vivo
title_fullStr Imaging, myeloid precursor immortalization, and genome editing for defining mechanisms of leukocyte recruitment in vivo
title_full_unstemmed Imaging, myeloid precursor immortalization, and genome editing for defining mechanisms of leukocyte recruitment in vivo
title_short Imaging, myeloid precursor immortalization, and genome editing for defining mechanisms of leukocyte recruitment in vivo
title_sort imaging, myeloid precursor immortalization, and genome editing for defining mechanisms of leukocyte recruitment in vivo
topic Research Paper
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5928898/
https://www.ncbi.nlm.nih.gov/pubmed/29721088
http://dx.doi.org/10.7150/thno.23632
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