PHD-2 Suppression in Mesenchymal Stromal Cells Enhances Wound Healing

Sae Hee Ko; Allison C. Nauta; Shane D. Morrison; Michael S. Hu; Andrew S. Zimmermann; Michael T. Chung; Jason P. Glotzbach; Victor W. Wong; Graham G. Walmsley; H. Peter Lorenz; Denise A. Chan; Geoffrey C. Gurtner; Amato J. Giaccia; Michael T. Longaker

doi:10.1097/PRS.0000000000003959

PHD-2 Suppression in Mesenchymal Stromal Cells Enhances Wound Healing

Sae Hee Ko, Allison C. Nauta, Shane D. Morrison, Michael S. Hu, Andrew S. Zimmermann, Michael T. Chung, Jason P. Glotzbach, Victor W. Wong, Graham G. Walmsley, H. Peter Lorenz, Denise A. Chan, Geoffrey C. Gurtner, Amato J. Giaccia, Michael T. Longaker

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

17 Scopus citations

Abstract

Background: Cell therapy with mesenchymal stromal cells is a promising strategy for tissue repair. Restoration of blood flow to ischemic tissues is a key step in wound repair, and mesenchymal stromal cells have been shown to be proangiogenic. Angiogenesis is critically regulated by the hypoxia-inducible factor (HIF) superfamily, consisting of transcription factors targeted for degradation by prolyl hydroxylase domain (PHD)-2. The aim of this study was to enhance the proangiogenic capability of mesenchymal stromal cells and to use these modified cells to promote wound healing. Methods: Mesenchymal stromal cells harvested from mouse bone marrow were transduced with short hairpin RNA (shRNA) against PHD-2; control cells were transduced with scrambled shRNA (shScramble) construct. Gene expression quantification, human umbilical vein endothelial cell tube formation assays, and wound healing assays were used to assess the effect of PHD knockdown mesenchymal stromal cells on wound healing dynamics. Results: PHD-2 knockdown mesenchymal stromal cells overexpressed HIF-1α and multiple angiogenic factors compared to control (p < 0.05). Human umbilical vein endothelial cells treated with conditioned medium from PHD-2 knockdown mesenchymal stromal cells exhibited increased formation of capillary-like structures and enhanced migration compared with human umbilical vein endothelial cells treated with conditioned medium from shScramble-transduced mesenchymal stromal cells (p < 0.05). Wounds treated with PHD-2 knockdown mesenchymal stromal cells healed at a significantly accelerated rate compared with wounds treated with shScramble mesenchymal stromal cells (p < 0.05). Histologic studies revealed increased blood vessel density and increased cellularity in the wounds treated with PHD-2 knockdown mesenchymal stromal cells (p < 0.05). Conclusions: Silencing PHD-2 in mesenchymal stromal cells augments their proangiogenic potential in wound healing therapy. This effect appears to be mediated by overexpression of HIF family transcription factors and up-regulation of multiple downstream angiogenic factors.

Original language	English (US)
Pages (from-to)	55e-67e
Journal	Plastic and reconstructive surgery
Volume	141
Issue number	1
DOIs	https://doi.org/10.1097/PRS.0000000000003959
State	Published - Jan 1 2018

ASJC Scopus subject areas

Surgery

Access to Document

10.1097/PRS.0000000000003959

Cite this

Ko, S. H., Nauta, A. C., Morrison, S. D., Hu, M. S., Zimmermann, A. S., Chung, M. T., Glotzbach, J. P., Wong, V. W., Walmsley, G. G., Peter Lorenz, H., Chan, D. A., Gurtner, G. C., Giaccia, A. J., & Longaker, M. T. (2018). PHD-2 Suppression in Mesenchymal Stromal Cells Enhances Wound Healing. Plastic and reconstructive surgery, 141(1), 55e-67e. https://doi.org/10.1097/PRS.0000000000003959

@article{11440638d4214130a4bd06660d60a201,

title = "PHD-2 Suppression in Mesenchymal Stromal Cells Enhances Wound Healing",

abstract = "Background: Cell therapy with mesenchymal stromal cells is a promising strategy for tissue repair. Restoration of blood flow to ischemic tissues is a key step in wound repair, and mesenchymal stromal cells have been shown to be proangiogenic. Angiogenesis is critically regulated by the hypoxia-inducible factor (HIF) superfamily, consisting of transcription factors targeted for degradation by prolyl hydroxylase domain (PHD)-2. The aim of this study was to enhance the proangiogenic capability of mesenchymal stromal cells and to use these modified cells to promote wound healing. Methods: Mesenchymal stromal cells harvested from mouse bone marrow were transduced with short hairpin RNA (shRNA) against PHD-2; control cells were transduced with scrambled shRNA (shScramble) construct. Gene expression quantification, human umbilical vein endothelial cell tube formation assays, and wound healing assays were used to assess the effect of PHD knockdown mesenchymal stromal cells on wound healing dynamics. Results: PHD-2 knockdown mesenchymal stromal cells overexpressed HIF-1α and multiple angiogenic factors compared to control (p < 0.05). Human umbilical vein endothelial cells treated with conditioned medium from PHD-2 knockdown mesenchymal stromal cells exhibited increased formation of capillary-like structures and enhanced migration compared with human umbilical vein endothelial cells treated with conditioned medium from shScramble-transduced mesenchymal stromal cells (p < 0.05). Wounds treated with PHD-2 knockdown mesenchymal stromal cells healed at a significantly accelerated rate compared with wounds treated with shScramble mesenchymal stromal cells (p < 0.05). Histologic studies revealed increased blood vessel density and increased cellularity in the wounds treated with PHD-2 knockdown mesenchymal stromal cells (p < 0.05). Conclusions: Silencing PHD-2 in mesenchymal stromal cells augments their proangiogenic potential in wound healing therapy. This effect appears to be mediated by overexpression of HIF family transcription factors and up-regulation of multiple downstream angiogenic factors.",

author = "Ko, {Sae Hee} and Nauta, {Allison C.} and Morrison, {Shane D.} and Hu, {Michael S.} and Zimmermann, {Andrew S.} and Chung, {Michael T.} and Glotzbach, {Jason P.} and Wong, {Victor W.} and Walmsley, {Graham G.} and {Peter Lorenz}, H. and Chan, {Denise A.} and Gurtner, {Geoffrey C.} and Giaccia, {Amato J.} and Longaker, {Michael T.}",

note = "Publisher Copyright: Copyright {\textcopyright} 2017 by the American Society of Plastic Surgeons.",

year = "2018",

month = jan,

day = "1",

doi = "10.1097/PRS.0000000000003959",

language = "English (US)",

volume = "141",

pages = "55e--67e",

journal = "Plastic and reconstructive surgery",

issn = "0032-1052",

publisher = "Lippincott Williams and Wilkins",

number = "1",

}

TY - JOUR

T1 - PHD-2 Suppression in Mesenchymal Stromal Cells Enhances Wound Healing

AU - Ko, Sae Hee

AU - Nauta, Allison C.

AU - Morrison, Shane D.

AU - Hu, Michael S.

AU - Zimmermann, Andrew S.

AU - Chung, Michael T.

AU - Glotzbach, Jason P.

AU - Wong, Victor W.

AU - Walmsley, Graham G.

AU - Peter Lorenz, H.

AU - Chan, Denise A.

AU - Gurtner, Geoffrey C.

AU - Giaccia, Amato J.

AU - Longaker, Michael T.

PY - 2018/1/1

Y1 - 2018/1/1

N2 - Background: Cell therapy with mesenchymal stromal cells is a promising strategy for tissue repair. Restoration of blood flow to ischemic tissues is a key step in wound repair, and mesenchymal stromal cells have been shown to be proangiogenic. Angiogenesis is critically regulated by the hypoxia-inducible factor (HIF) superfamily, consisting of transcription factors targeted for degradation by prolyl hydroxylase domain (PHD)-2. The aim of this study was to enhance the proangiogenic capability of mesenchymal stromal cells and to use these modified cells to promote wound healing. Methods: Mesenchymal stromal cells harvested from mouse bone marrow were transduced with short hairpin RNA (shRNA) against PHD-2; control cells were transduced with scrambled shRNA (shScramble) construct. Gene expression quantification, human umbilical vein endothelial cell tube formation assays, and wound healing assays were used to assess the effect of PHD knockdown mesenchymal stromal cells on wound healing dynamics. Results: PHD-2 knockdown mesenchymal stromal cells overexpressed HIF-1α and multiple angiogenic factors compared to control (p < 0.05). Human umbilical vein endothelial cells treated with conditioned medium from PHD-2 knockdown mesenchymal stromal cells exhibited increased formation of capillary-like structures and enhanced migration compared with human umbilical vein endothelial cells treated with conditioned medium from shScramble-transduced mesenchymal stromal cells (p < 0.05). Wounds treated with PHD-2 knockdown mesenchymal stromal cells healed at a significantly accelerated rate compared with wounds treated with shScramble mesenchymal stromal cells (p < 0.05). Histologic studies revealed increased blood vessel density and increased cellularity in the wounds treated with PHD-2 knockdown mesenchymal stromal cells (p < 0.05). Conclusions: Silencing PHD-2 in mesenchymal stromal cells augments their proangiogenic potential in wound healing therapy. This effect appears to be mediated by overexpression of HIF family transcription factors and up-regulation of multiple downstream angiogenic factors.

AB - Background: Cell therapy with mesenchymal stromal cells is a promising strategy for tissue repair. Restoration of blood flow to ischemic tissues is a key step in wound repair, and mesenchymal stromal cells have been shown to be proangiogenic. Angiogenesis is critically regulated by the hypoxia-inducible factor (HIF) superfamily, consisting of transcription factors targeted for degradation by prolyl hydroxylase domain (PHD)-2. The aim of this study was to enhance the proangiogenic capability of mesenchymal stromal cells and to use these modified cells to promote wound healing. Methods: Mesenchymal stromal cells harvested from mouse bone marrow were transduced with short hairpin RNA (shRNA) against PHD-2; control cells were transduced with scrambled shRNA (shScramble) construct. Gene expression quantification, human umbilical vein endothelial cell tube formation assays, and wound healing assays were used to assess the effect of PHD knockdown mesenchymal stromal cells on wound healing dynamics. Results: PHD-2 knockdown mesenchymal stromal cells overexpressed HIF-1α and multiple angiogenic factors compared to control (p < 0.05). Human umbilical vein endothelial cells treated with conditioned medium from PHD-2 knockdown mesenchymal stromal cells exhibited increased formation of capillary-like structures and enhanced migration compared with human umbilical vein endothelial cells treated with conditioned medium from shScramble-transduced mesenchymal stromal cells (p < 0.05). Wounds treated with PHD-2 knockdown mesenchymal stromal cells healed at a significantly accelerated rate compared with wounds treated with shScramble mesenchymal stromal cells (p < 0.05). Histologic studies revealed increased blood vessel density and increased cellularity in the wounds treated with PHD-2 knockdown mesenchymal stromal cells (p < 0.05). Conclusions: Silencing PHD-2 in mesenchymal stromal cells augments their proangiogenic potential in wound healing therapy. This effect appears to be mediated by overexpression of HIF family transcription factors and up-regulation of multiple downstream angiogenic factors.

UR - http://www.scopus.com/inward/record.url?scp=85039745061&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85039745061&partnerID=8YFLogxK

U2 - 10.1097/PRS.0000000000003959

DO - 10.1097/PRS.0000000000003959

M3 - Article

C2 - 29280872

AN - SCOPUS:85039745061

SN - 0032-1052

VL - 141

SP - 55e-67e

JO - Plastic and reconstructive surgery

JF - Plastic and reconstructive surgery

IS - 1

ER -

PHD-2 Suppression in Mesenchymal Stromal Cells Enhances Wound Healing

Abstract

ASJC Scopus subject areas

Access to Document

Other files and links

Fingerprint

Cite this