Selective ROCK Inhibitor Enhances Blood Flow Recovery after Hindlimb Ischemia

<p dir="ltr">The impairment in microvascular network formation could delay the restoration of blood flow after acute limb ischemia. A high-content screen of a GSK-published kinase inhibitor library identified a set of ROCK inhibitor hits enhancing endothelial network formation. Subse...

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محفوظ في:
التفاصيل البيبلوغرافية
المؤلف الرئيسي: Hend Salah Fayed (17732622) (author)
مؤلفون آخرون: Mouayad Zuheir Bakleh (19450432) (author), Jasni Viralippurath Ashraf (18518943) (author), Alison Howarth (7356479) (author), Daniel Ebner (5556488) (author), Ayman Al Haj Zen (16614801) (author)
منشور في: 2023
الموضوعات:
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_version_ 1864513507658039296
author Hend Salah Fayed (17732622)
author2 Mouayad Zuheir Bakleh (19450432)
Jasni Viralippurath Ashraf (18518943)
Alison Howarth (7356479)
Daniel Ebner (5556488)
Ayman Al Haj Zen (16614801)
author2_role author
author
author
author
author
author_facet Hend Salah Fayed (17732622)
Mouayad Zuheir Bakleh (19450432)
Jasni Viralippurath Ashraf (18518943)
Alison Howarth (7356479)
Daniel Ebner (5556488)
Ayman Al Haj Zen (16614801)
author_role author
dc.creator.none.fl_str_mv Hend Salah Fayed (17732622)
Mouayad Zuheir Bakleh (19450432)
Jasni Viralippurath Ashraf (18518943)
Alison Howarth (7356479)
Daniel Ebner (5556488)
Ayman Al Haj Zen (16614801)
dc.date.none.fl_str_mv 2023-09-22T09:00:00Z
dc.identifier.none.fl_str_mv 10.3390/ijms241914410
dc.relation.none.fl_str_mv https://figshare.com/articles/journal_contribution/Selective_ROCK_Inhibitor_Enhances_Blood_Flow_Recovery_after_Hindlimb_Ischemia/26788333
dc.rights.none.fl_str_mv CC BY 4.0
info:eu-repo/semantics/openAccess
dc.subject.none.fl_str_mv Biological sciences
Biochemistry and cell biology
Biomedical and clinical sciences
Pharmacology and pharmaceutical sciences
high content screening
kinase inhibitors
ROCK inhibitor
blood flow
limb ischemia
dc.title.none.fl_str_mv Selective ROCK Inhibitor Enhances Blood Flow Recovery after Hindlimb Ischemia
dc.type.none.fl_str_mv Text
Journal contribution
info:eu-repo/semantics/publishedVersion
text
contribution to journal
description <p dir="ltr">The impairment in microvascular network formation could delay the restoration of blood flow after acute limb ischemia. A high-content screen of a GSK-published kinase inhibitor library identified a set of ROCK inhibitor hits enhancing endothelial network formation. Subsequent kinase activity profiling against a panel of 224 protein kinases showed that two indazole-based ROCK inhibitor hits exhibited high selectivity for ROCK1 and ROCK2 isoforms compared to other ROCK inhibitors. One of the chemical entities, GSK429286, was selected for follow-up studies. We found that GSK429286 was ten times more potent in enhancing endothelial tube formation than Fasudil, a classic ROCK inhibitor. ROCK1 inhibition by RNAi phenocopied the angiogenic phenotype of the GSK429286 compound. Using an organotypic angiogenesis co-culture assay, we showed that GSK429286 formed a dense vascular network with thicker endothelial tubes. Next, mice received either vehicle or GSK429286 (10 mg/kg i.p.) for seven days after hindlimb ischemia induction. As assessed by laser speckle contrast imaging, GSK429286 potentiated blood flow recovery after ischemia induction. At the histological level, we found that GSK429286 significantly increased the size of new microvessels in the regenerating areas of ischemic muscles compared with vehicle-treated ones. Our findings reveal that selective ROCK inhibitors have in vitro pro-angiogenic properties and therapeutic potential to restore blood flow in limb ischemia. </p><h2>Other Information</h2><p dir="ltr">Published in: International Journal of Molecular Sciences<br>License: <a href="https://creativecommons.org/licenses/by/4.0/" target="_blank">https://creativecommons.org/licenses/by/4.0/</a><br>See article on publisher's website: <a href="https://dx.doi.org/10.3390/ijms241914410" target="_blank">https://dx.doi.org/10.3390/ijms241914410</a></p>
eu_rights_str_mv openAccess
id Manara2_fb9528a825024e4856a354cdf4a706fb
identifier_str_mv 10.3390/ijms241914410
network_acronym_str Manara2
network_name_str Manara2
oai_identifier_str oai:figshare.com:article/26788333
publishDate 2023
repository.mail.fl_str_mv
repository.name.fl_str_mv
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rights_invalid_str_mv CC BY 4.0
spelling Selective ROCK Inhibitor Enhances Blood Flow Recovery after Hindlimb IschemiaHend Salah Fayed (17732622)Mouayad Zuheir Bakleh (19450432)Jasni Viralippurath Ashraf (18518943)Alison Howarth (7356479)Daniel Ebner (5556488)Ayman Al Haj Zen (16614801)Biological sciencesBiochemistry and cell biologyBiomedical and clinical sciencesPharmacology and pharmaceutical scienceshigh content screeningkinase inhibitorsROCK inhibitorblood flowlimb ischemia<p dir="ltr">The impairment in microvascular network formation could delay the restoration of blood flow after acute limb ischemia. A high-content screen of a GSK-published kinase inhibitor library identified a set of ROCK inhibitor hits enhancing endothelial network formation. Subsequent kinase activity profiling against a panel of 224 protein kinases showed that two indazole-based ROCK inhibitor hits exhibited high selectivity for ROCK1 and ROCK2 isoforms compared to other ROCK inhibitors. One of the chemical entities, GSK429286, was selected for follow-up studies. We found that GSK429286 was ten times more potent in enhancing endothelial tube formation than Fasudil, a classic ROCK inhibitor. ROCK1 inhibition by RNAi phenocopied the angiogenic phenotype of the GSK429286 compound. Using an organotypic angiogenesis co-culture assay, we showed that GSK429286 formed a dense vascular network with thicker endothelial tubes. Next, mice received either vehicle or GSK429286 (10 mg/kg i.p.) for seven days after hindlimb ischemia induction. As assessed by laser speckle contrast imaging, GSK429286 potentiated blood flow recovery after ischemia induction. At the histological level, we found that GSK429286 significantly increased the size of new microvessels in the regenerating areas of ischemic muscles compared with vehicle-treated ones. Our findings reveal that selective ROCK inhibitors have in vitro pro-angiogenic properties and therapeutic potential to restore blood flow in limb ischemia. </p><h2>Other Information</h2><p dir="ltr">Published in: International Journal of Molecular Sciences<br>License: <a href="https://creativecommons.org/licenses/by/4.0/" target="_blank">https://creativecommons.org/licenses/by/4.0/</a><br>See article on publisher's website: <a href="https://dx.doi.org/10.3390/ijms241914410" target="_blank">https://dx.doi.org/10.3390/ijms241914410</a></p>2023-09-22T09:00:00ZTextJournal contributioninfo:eu-repo/semantics/publishedVersiontextcontribution to journal10.3390/ijms241914410https://figshare.com/articles/journal_contribution/Selective_ROCK_Inhibitor_Enhances_Blood_Flow_Recovery_after_Hindlimb_Ischemia/26788333CC BY 4.0info:eu-repo/semantics/openAccessoai:figshare.com:article/267883332023-09-22T09:00:00Z
spellingShingle Selective ROCK Inhibitor Enhances Blood Flow Recovery after Hindlimb Ischemia
Hend Salah Fayed (17732622)
Biological sciences
Biochemistry and cell biology
Biomedical and clinical sciences
Pharmacology and pharmaceutical sciences
high content screening
kinase inhibitors
ROCK inhibitor
blood flow
limb ischemia
status_str publishedVersion
title Selective ROCK Inhibitor Enhances Blood Flow Recovery after Hindlimb Ischemia
title_full Selective ROCK Inhibitor Enhances Blood Flow Recovery after Hindlimb Ischemia
title_fullStr Selective ROCK Inhibitor Enhances Blood Flow Recovery after Hindlimb Ischemia
title_full_unstemmed Selective ROCK Inhibitor Enhances Blood Flow Recovery after Hindlimb Ischemia
title_short Selective ROCK Inhibitor Enhances Blood Flow Recovery after Hindlimb Ischemia
title_sort Selective ROCK Inhibitor Enhances Blood Flow Recovery after Hindlimb Ischemia
topic Biological sciences
Biochemistry and cell biology
Biomedical and clinical sciences
Pharmacology and pharmaceutical sciences
high content screening
kinase inhibitors
ROCK inhibitor
blood flow
limb ischemia