The interaction of ceramide 1-phosphate with group IVA cytosolic phospholipase A2 coordinates acute wound healing and repair

H. Patrick MacKnight; Daniel J. Stephenson; L. Alexis Hoeferlin; Savannah D. Benusa; James T. DeLigio; Kenneth D.  Maus; Anika N. Ali; Jennifer S. Wayne; Margaret A. Park; Edward H Hinchcliffe; Rhoderick E Brown; John J. Ryan; Robert F. Diegelmann; Charles E. Chalfant

doi:10.1126/scisignal.aav5918

The interaction of ceramide 1-phosphate with group IVA cytosolic phospholipase A2 coordinates acute wound healing and repair

H. Patrick MacKnight, Daniel J. Stephenson, L. Alexis Hoeferlin, Savannah D. Benusa, James T. DeLigio, Kenneth D. Maus, Anika N. Ali, Jennifer S. Wayne, Margaret A. Park, Edward H Hinchcliffe, Rhoderick E Brown, John J. Ryan, Robert F. Diegelmann, Charles E. Chalfant

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28 Scopus citations

Abstract

The sphingolipid ceramide 1-phosphate (C1P) directly binds to and activates group IVA cytosolic phospholipase A ₂ (cPLA ₂α) to stimulate the production of eicosanoids. Because eicosanoids are important in wound healing, we examined the repair of skin wounds in knockout (KO) mice lacking cPLA ₂α and in knock-in (KI) mice in which endogenous cPLA ₂α was replaced with a mutant form having an ablated C1P interaction site. Wound closure rate was not affected in the KO or KI mice, but wound maturation was enhanced in the KI mice compared to that in wild-type controls. Wounds in KI mice displayed increased infiltration of dermal fibroblasts into the wound environment, increased wound tensile strength, and a higher ratio of type I:type III collagen. In vitro, primary dermal fibroblasts (pDFs) from KI mice showed substantially increased collagen deposition and migration velocity compared to pDFs from wild-type and KO mice. KI mice also showed an altered eicosanoid profile of reduced proinflammatory prostaglandins (PGE ₂ and TXB ₂) and an increased abundance of certain hydroxyeicosatetraenoic acid (HETE) species. Specifically, an increase in 5-HETE enhanced dermal fibroblast migration and collagen deposition. This gain-of-function role for the mutant cPLA ₂α was also linked to the relocalization of cPLA ₂α and 5-HETE biosynthetic enzymes to the cytoplasm and cytoplasmic vesicles. These findings demonstrate the regulation of key wound-healing mechanisms in vivo by a defined protein-lipid interaction and provide insights into the roles that cPLA ₂α and eicosanoids play in orchestrating wound repair.

Original language	English (US)
Article number	eaav5918
Number of pages	17
Journal	Science Signaling
Volume	12
Issue number	610
DOIs	https://doi.org/10.1126/scisignal.aav5918
State	Published - Dec 3 2019

Bibliographical note

Funding Information:
We would like to thank J. Ryan and laboratory group in the Biology Department at VCU for use of laboratory space to finish experiments integral to the completion of this manuscript This work was supported by research grants from the Veteran?s Administration [VA Merit Review, I BX001792 (C.E.C.) and a Research Career Scientist Award, 13F-RCS-002 (C.E.C.)] and from the NIH via HL125353 (C.E.C., R.E.B., and E.H.H.), HD087198 and RR031535 (C.E.C.), R01138495 (J.J.R.), and F32Al108088 (L.A.H.). The contents of this manuscript do not represent the views of the Department of Veterans Affairs or the U.S. government. Microscopy was performed at the VCU Microscopy Facility, supported, in part, by funding from NIH-NCI Cancer Center Support Grant P30 CA016059.

Publisher Copyright:
Copyright © 2019 The Authors,

PubMed: MeSH publication types

Research Support, U.S. Gov't, Non-P.H.S.
Journal Article
Research Support, N.I.H., Extramural

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MacKnight, H. P., Stephenson, D. J., Hoeferlin, L. A., Benusa, S. D., DeLigio, J. T., Maus, K. D., Ali, A. N., Wayne, J. S., Park, M. A., Hinchcliffe, E. H., Brown, R. E., Ryan, J. J., Diegelmann, R. F., & Chalfant, C. E. (2019). The interaction of ceramide 1-phosphate with group IVA cytosolic phospholipase A2 coordinates acute wound healing and repair. Science Signaling, 12(610), Article eaav5918. https://doi.org/10.1126/scisignal.aav5918

MacKnight, HP, Stephenson, DJ, Hoeferlin, LA, Benusa, SD, DeLigio, JT, Maus, KD, Ali, AN, Wayne, JS, Park, MA, Hinchcliffe, EH, Brown, RE, Ryan, JJ, Diegelmann, RF & Chalfant, CE 2019, 'The interaction of ceramide 1-phosphate with group IVA cytosolic phospholipase A2 coordinates acute wound healing and repair', Science Signaling, vol. 12, no. 610, eaav5918. https://doi.org/10.1126/scisignal.aav5918

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title = "The interaction of ceramide 1-phosphate with group IVA cytosolic phospholipase A2 coordinates acute wound healing and repair",

abstract = "The sphingolipid ceramide 1-phosphate (C1P) directly binds to and activates group IVA cytosolic phospholipase A 2 (cPLA 2α) to stimulate the production of eicosanoids. Because eicosanoids are important in wound healing, we examined the repair of skin wounds in knockout (KO) mice lacking cPLA 2α and in knock-in (KI) mice in which endogenous cPLA 2α was replaced with a mutant form having an ablated C1P interaction site. Wound closure rate was not affected in the KO or KI mice, but wound maturation was enhanced in the KI mice compared to that in wild-type controls. Wounds in KI mice displayed increased infiltration of dermal fibroblasts into the wound environment, increased wound tensile strength, and a higher ratio of type I:type III collagen. In vitro, primary dermal fibroblasts (pDFs) from KI mice showed substantially increased collagen deposition and migration velocity compared to pDFs from wild-type and KO mice. KI mice also showed an altered eicosanoid profile of reduced proinflammatory prostaglandins (PGE 2 and TXB 2) and an increased abundance of certain hydroxyeicosatetraenoic acid (HETE) species. Specifically, an increase in 5-HETE enhanced dermal fibroblast migration and collagen deposition. This gain-of-function role for the mutant cPLA 2α was also linked to the relocalization of cPLA 2α and 5-HETE biosynthetic enzymes to the cytoplasm and cytoplasmic vesicles. These findings demonstrate the regulation of key wound-healing mechanisms in vivo by a defined protein-lipid interaction and provide insights into the roles that cPLA 2α and eicosanoids play in orchestrating wound repair. ",

author = "MacKnight, {H. Patrick} and Stephenson, {Daniel J.} and Hoeferlin, {L. Alexis} and Benusa, {Savannah D.} and DeLigio, {James T.} and Maus, {Kenneth D.} and Ali, {Anika N.} and Wayne, {Jennifer S.} and Park, {Margaret A.} and Hinchcliffe, {Edward H} and Brown, {Rhoderick E} and Ryan, {John J.} and Diegelmann, {Robert F.} and Chalfant, {Charles E.}",

note = "Funding Information: We would like to thank J. Ryan and laboratory group in the Biology Department at VCU for use of laboratory space to finish experiments integral to the completion of this manuscript This work was supported by research grants from the Veteran?s Administration [VA Merit Review, I BX001792 (C.E.C.) and a Research Career Scientist Award, 13F-RCS-002 (C.E.C.)] and from the NIH via HL125353 (C.E.C., R.E.B., and E.H.H.), HD087198 and RR031535 (C.E.C.), R01138495 (J.J.R.), and F32Al108088 (L.A.H.). The contents of this manuscript do not represent the views of the Department of Veterans Affairs or the U.S. government. Microscopy was performed at the VCU Microscopy Facility, supported, in part, by funding from NIH-NCI Cancer Center Support Grant P30 CA016059. Publisher Copyright: Copyright {\textcopyright} 2019 The Authors,",

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T1 - The interaction of ceramide 1-phosphate with group IVA cytosolic phospholipase A2 coordinates acute wound healing and repair

AU - MacKnight, H. Patrick

AU - Stephenson, Daniel J.

AU - Hoeferlin, L. Alexis

AU - Benusa, Savannah D.

AU - DeLigio, James T.

AU - Maus, Kenneth D.

AU - Ali, Anika N.

AU - Wayne, Jennifer S.

AU - Park, Margaret A.

AU - Hinchcliffe, Edward H

AU - Brown, Rhoderick E

AU - Ryan, John J.

AU - Diegelmann, Robert F.

AU - Chalfant, Charles E.

N1 - Funding Information: We would like to thank J. Ryan and laboratory group in the Biology Department at VCU for use of laboratory space to finish experiments integral to the completion of this manuscript This work was supported by research grants from the Veteran?s Administration [VA Merit Review, I BX001792 (C.E.C.) and a Research Career Scientist Award, 13F-RCS-002 (C.E.C.)] and from the NIH via HL125353 (C.E.C., R.E.B., and E.H.H.), HD087198 and RR031535 (C.E.C.), R01138495 (J.J.R.), and F32Al108088 (L.A.H.). The contents of this manuscript do not represent the views of the Department of Veterans Affairs or the U.S. government. Microscopy was performed at the VCU Microscopy Facility, supported, in part, by funding from NIH-NCI Cancer Center Support Grant P30 CA016059. Publisher Copyright: Copyright © 2019 The Authors,

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N2 - The sphingolipid ceramide 1-phosphate (C1P) directly binds to and activates group IVA cytosolic phospholipase A 2 (cPLA 2α) to stimulate the production of eicosanoids. Because eicosanoids are important in wound healing, we examined the repair of skin wounds in knockout (KO) mice lacking cPLA 2α and in knock-in (KI) mice in which endogenous cPLA 2α was replaced with a mutant form having an ablated C1P interaction site. Wound closure rate was not affected in the KO or KI mice, but wound maturation was enhanced in the KI mice compared to that in wild-type controls. Wounds in KI mice displayed increased infiltration of dermal fibroblasts into the wound environment, increased wound tensile strength, and a higher ratio of type I:type III collagen. In vitro, primary dermal fibroblasts (pDFs) from KI mice showed substantially increased collagen deposition and migration velocity compared to pDFs from wild-type and KO mice. KI mice also showed an altered eicosanoid profile of reduced proinflammatory prostaglandins (PGE 2 and TXB 2) and an increased abundance of certain hydroxyeicosatetraenoic acid (HETE) species. Specifically, an increase in 5-HETE enhanced dermal fibroblast migration and collagen deposition. This gain-of-function role for the mutant cPLA 2α was also linked to the relocalization of cPLA 2α and 5-HETE biosynthetic enzymes to the cytoplasm and cytoplasmic vesicles. These findings demonstrate the regulation of key wound-healing mechanisms in vivo by a defined protein-lipid interaction and provide insights into the roles that cPLA 2α and eicosanoids play in orchestrating wound repair.

AB - The sphingolipid ceramide 1-phosphate (C1P) directly binds to and activates group IVA cytosolic phospholipase A 2 (cPLA 2α) to stimulate the production of eicosanoids. Because eicosanoids are important in wound healing, we examined the repair of skin wounds in knockout (KO) mice lacking cPLA 2α and in knock-in (KI) mice in which endogenous cPLA 2α was replaced with a mutant form having an ablated C1P interaction site. Wound closure rate was not affected in the KO or KI mice, but wound maturation was enhanced in the KI mice compared to that in wild-type controls. Wounds in KI mice displayed increased infiltration of dermal fibroblasts into the wound environment, increased wound tensile strength, and a higher ratio of type I:type III collagen. In vitro, primary dermal fibroblasts (pDFs) from KI mice showed substantially increased collagen deposition and migration velocity compared to pDFs from wild-type and KO mice. KI mice also showed an altered eicosanoid profile of reduced proinflammatory prostaglandins (PGE 2 and TXB 2) and an increased abundance of certain hydroxyeicosatetraenoic acid (HETE) species. Specifically, an increase in 5-HETE enhanced dermal fibroblast migration and collagen deposition. This gain-of-function role for the mutant cPLA 2α was also linked to the relocalization of cPLA 2α and 5-HETE biosynthetic enzymes to the cytoplasm and cytoplasmic vesicles. These findings demonstrate the regulation of key wound-healing mechanisms in vivo by a defined protein-lipid interaction and provide insights into the roles that cPLA 2α and eicosanoids play in orchestrating wound repair.

U2 - 10.1126/scisignal.aav5918

DO - 10.1126/scisignal.aav5918

M3 - Article

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SN - 1937-9145

VL - 12

JO - Science Signaling

JF - Science Signaling

IS - 610

M1 - eaav5918

ER -

The interaction of ceramide 1-phosphate with group IVA cytosolic phospholipase A2 coordinates acute wound healing and repair

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