Inhibition of Hif1α prevents both trauma-induced and genetic heterotopic ossification

Shailesh Agarwal, Shawn Loder, Cameron Brownley, David Cholok, Laura Mangiavini, John Li, Christopher Breuler, Hsiao H. Sung, Shuli Li, Kavitha Ranganathan, Joshua Peterson, Ronald Tompkins, David Herndon, Wenzhong Xiao, Dolrudee Jumlongras, Bjorn R. Olsen, Thomas A. Davis, Yuji Mishina, Ernestina Schipani, Benjamin Levi

Research output: Contribution to journalArticle

Abstract

Pathologic extraskeletal bone formation, or heterotopic ossification (HO), occurs following mechanical trauma, burns, orthopedic operations, and in patients with hyperactivating mutations of the type I bone morphogenetic protein receptor ACVR1 (Activin type 1 receptor). Extraskeletal bone forms through an endochondral process with a cartilage intermediary prompting the hypothesis that hypoxic signaling present during cartilage formation drives HO development and that HO precursor cells derive from a mesenchymal lineage as defined by Paired related homeobox 1 (Prx). Here we demonstrate that Hypoxia inducible factor-1α (Hif1α), a key mediator of cellular adaptation to hypoxia, is highly expressed and active in three separate mouse models: trauma-induced, genetic, and a hybrid model of genetic and trauma-induced HO. In each of these models, Hif1α expression coincides with the expression of master transcription factor of cartilage, Sox9 [(sex determining region Y)- box 9]. Pharmacologic inhibition of Hif1α using PX-478 or rapamycin significantly decreased or inhibited extraskeletal bone formation. Importantly, de novo soft-tissue HO was eliminated or significantly diminished in treated mice. Lineage-tracing mice demonstrate that cells forming HO belong to the Prx lineage. Burn/tenotomy performed in lineage-specific Hif1α knockout mice (Prx-Cre/Hif1αfl:fl) resulted in substantially decreased HO, and again lack of de novo soft-tissue HO. Genetic loss of Hif1α in mesenchymal cells marked by Prx-cre prevents the formation of the mesenchymal condensations as shown by routine histology and immunostaining for Sox9 and PDGFRα. Pharmacologic inhibition of Hif1α had a similar effect on mesenchymal condensation development. Our findings indicate that Hif1α represents a promising target to prevent and treat pathologic extraskeletal bone.

Original languageEnglish (US)
Pages (from-to)E338-E347
JournalProceedings of the National Academy of Sciences of the United States of America
Volume113
Issue number3
DOIs
StatePublished - Jan 19 2016
Externally publishedYes

Fingerprint

Heterotopic Ossification
Hypoxia-Inducible Factor 1
Wounds and Injuries
Cartilage
Burns
Osteogenesis
Type I Activin Receptors
Type I Bone Morphogenetic Protein Receptors
Tenotomy
Bone and Bones
Homeobox Genes
Genetic Models
Sirolimus
Knockout Mice
Orthopedics
Histology
Transcription Factors
Mutation

Keywords

  • Cartilage
  • Heterotopic ossification
  • HIF1α
  • Mesenchymal condensation
  • Prx

ASJC Scopus subject areas

  • General

Cite this

Inhibition of Hif1α prevents both trauma-induced and genetic heterotopic ossification. / Agarwal, Shailesh; Loder, Shawn; Brownley, Cameron; Cholok, David; Mangiavini, Laura; Li, John; Breuler, Christopher; Sung, Hsiao H.; Li, Shuli; Ranganathan, Kavitha; Peterson, Joshua; Tompkins, Ronald; Herndon, David; Xiao, Wenzhong; Jumlongras, Dolrudee; Olsen, Bjorn R.; Davis, Thomas A.; Mishina, Yuji; Schipani, Ernestina; Levi, Benjamin.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 113, No. 3, 19.01.2016, p. E338-E347.

Research output: Contribution to journalArticle

Agarwal, S, Loder, S, Brownley, C, Cholok, D, Mangiavini, L, Li, J, Breuler, C, Sung, HH, Li, S, Ranganathan, K, Peterson, J, Tompkins, R, Herndon, D, Xiao, W, Jumlongras, D, Olsen, BR, Davis, TA, Mishina, Y, Schipani, E & Levi, B 2016, 'Inhibition of Hif1α prevents both trauma-induced and genetic heterotopic ossification', Proceedings of the National Academy of Sciences of the United States of America, vol. 113, no. 3, pp. E338-E347. https://doi.org/10.1073/pnas.1515397113
Agarwal, Shailesh ; Loder, Shawn ; Brownley, Cameron ; Cholok, David ; Mangiavini, Laura ; Li, John ; Breuler, Christopher ; Sung, Hsiao H. ; Li, Shuli ; Ranganathan, Kavitha ; Peterson, Joshua ; Tompkins, Ronald ; Herndon, David ; Xiao, Wenzhong ; Jumlongras, Dolrudee ; Olsen, Bjorn R. ; Davis, Thomas A. ; Mishina, Yuji ; Schipani, Ernestina ; Levi, Benjamin. / Inhibition of Hif1α prevents both trauma-induced and genetic heterotopic ossification. In: Proceedings of the National Academy of Sciences of the United States of America. 2016 ; Vol. 113, No. 3. pp. E338-E347.
@article{501e234e483e4a8697a3daf277864952,
title = "Inhibition of Hif1α prevents both trauma-induced and genetic heterotopic ossification",
abstract = "Pathologic extraskeletal bone formation, or heterotopic ossification (HO), occurs following mechanical trauma, burns, orthopedic operations, and in patients with hyperactivating mutations of the type I bone morphogenetic protein receptor ACVR1 (Activin type 1 receptor). Extraskeletal bone forms through an endochondral process with a cartilage intermediary prompting the hypothesis that hypoxic signaling present during cartilage formation drives HO development and that HO precursor cells derive from a mesenchymal lineage as defined by Paired related homeobox 1 (Prx). Here we demonstrate that Hypoxia inducible factor-1α (Hif1α), a key mediator of cellular adaptation to hypoxia, is highly expressed and active in three separate mouse models: trauma-induced, genetic, and a hybrid model of genetic and trauma-induced HO. In each of these models, Hif1α expression coincides with the expression of master transcription factor of cartilage, Sox9 [(sex determining region Y)- box 9]. Pharmacologic inhibition of Hif1α using PX-478 or rapamycin significantly decreased or inhibited extraskeletal bone formation. Importantly, de novo soft-tissue HO was eliminated or significantly diminished in treated mice. Lineage-tracing mice demonstrate that cells forming HO belong to the Prx lineage. Burn/tenotomy performed in lineage-specific Hif1α knockout mice (Prx-Cre/Hif1αfl:fl) resulted in substantially decreased HO, and again lack of de novo soft-tissue HO. Genetic loss of Hif1α in mesenchymal cells marked by Prx-cre prevents the formation of the mesenchymal condensations as shown by routine histology and immunostaining for Sox9 and PDGFRα. Pharmacologic inhibition of Hif1α had a similar effect on mesenchymal condensation development. Our findings indicate that Hif1α represents a promising target to prevent and treat pathologic extraskeletal bone.",
keywords = "Cartilage, Heterotopic ossification, HIF1α, Mesenchymal condensation, Prx",
author = "Shailesh Agarwal and Shawn Loder and Cameron Brownley and David Cholok and Laura Mangiavini and John Li and Christopher Breuler and Sung, {Hsiao H.} and Shuli Li and Kavitha Ranganathan and Joshua Peterson and Ronald Tompkins and David Herndon and Wenzhong Xiao and Dolrudee Jumlongras and Olsen, {Bjorn R.} and Davis, {Thomas A.} and Yuji Mishina and Ernestina Schipani and Benjamin Levi",
year = "2016",
month = "1",
day = "19",
doi = "10.1073/pnas.1515397113",
language = "English (US)",
volume = "113",
pages = "E338--E347",
journal = "Proceedings of the National Academy of Sciences of the United States of America",
issn = "0027-8424",
number = "3",

}

TY - JOUR

T1 - Inhibition of Hif1α prevents both trauma-induced and genetic heterotopic ossification

AU - Agarwal, Shailesh

AU - Loder, Shawn

AU - Brownley, Cameron

AU - Cholok, David

AU - Mangiavini, Laura

AU - Li, John

AU - Breuler, Christopher

AU - Sung, Hsiao H.

AU - Li, Shuli

AU - Ranganathan, Kavitha

AU - Peterson, Joshua

AU - Tompkins, Ronald

AU - Herndon, David

AU - Xiao, Wenzhong

AU - Jumlongras, Dolrudee

AU - Olsen, Bjorn R.

AU - Davis, Thomas A.

AU - Mishina, Yuji

AU - Schipani, Ernestina

AU - Levi, Benjamin

PY - 2016/1/19

Y1 - 2016/1/19

N2 - Pathologic extraskeletal bone formation, or heterotopic ossification (HO), occurs following mechanical trauma, burns, orthopedic operations, and in patients with hyperactivating mutations of the type I bone morphogenetic protein receptor ACVR1 (Activin type 1 receptor). Extraskeletal bone forms through an endochondral process with a cartilage intermediary prompting the hypothesis that hypoxic signaling present during cartilage formation drives HO development and that HO precursor cells derive from a mesenchymal lineage as defined by Paired related homeobox 1 (Prx). Here we demonstrate that Hypoxia inducible factor-1α (Hif1α), a key mediator of cellular adaptation to hypoxia, is highly expressed and active in three separate mouse models: trauma-induced, genetic, and a hybrid model of genetic and trauma-induced HO. In each of these models, Hif1α expression coincides with the expression of master transcription factor of cartilage, Sox9 [(sex determining region Y)- box 9]. Pharmacologic inhibition of Hif1α using PX-478 or rapamycin significantly decreased or inhibited extraskeletal bone formation. Importantly, de novo soft-tissue HO was eliminated or significantly diminished in treated mice. Lineage-tracing mice demonstrate that cells forming HO belong to the Prx lineage. Burn/tenotomy performed in lineage-specific Hif1α knockout mice (Prx-Cre/Hif1αfl:fl) resulted in substantially decreased HO, and again lack of de novo soft-tissue HO. Genetic loss of Hif1α in mesenchymal cells marked by Prx-cre prevents the formation of the mesenchymal condensations as shown by routine histology and immunostaining for Sox9 and PDGFRα. Pharmacologic inhibition of Hif1α had a similar effect on mesenchymal condensation development. Our findings indicate that Hif1α represents a promising target to prevent and treat pathologic extraskeletal bone.

AB - Pathologic extraskeletal bone formation, or heterotopic ossification (HO), occurs following mechanical trauma, burns, orthopedic operations, and in patients with hyperactivating mutations of the type I bone morphogenetic protein receptor ACVR1 (Activin type 1 receptor). Extraskeletal bone forms through an endochondral process with a cartilage intermediary prompting the hypothesis that hypoxic signaling present during cartilage formation drives HO development and that HO precursor cells derive from a mesenchymal lineage as defined by Paired related homeobox 1 (Prx). Here we demonstrate that Hypoxia inducible factor-1α (Hif1α), a key mediator of cellular adaptation to hypoxia, is highly expressed and active in three separate mouse models: trauma-induced, genetic, and a hybrid model of genetic and trauma-induced HO. In each of these models, Hif1α expression coincides with the expression of master transcription factor of cartilage, Sox9 [(sex determining region Y)- box 9]. Pharmacologic inhibition of Hif1α using PX-478 or rapamycin significantly decreased or inhibited extraskeletal bone formation. Importantly, de novo soft-tissue HO was eliminated or significantly diminished in treated mice. Lineage-tracing mice demonstrate that cells forming HO belong to the Prx lineage. Burn/tenotomy performed in lineage-specific Hif1α knockout mice (Prx-Cre/Hif1αfl:fl) resulted in substantially decreased HO, and again lack of de novo soft-tissue HO. Genetic loss of Hif1α in mesenchymal cells marked by Prx-cre prevents the formation of the mesenchymal condensations as shown by routine histology and immunostaining for Sox9 and PDGFRα. Pharmacologic inhibition of Hif1α had a similar effect on mesenchymal condensation development. Our findings indicate that Hif1α represents a promising target to prevent and treat pathologic extraskeletal bone.

KW - Cartilage

KW - Heterotopic ossification

KW - HIF1α

KW - Mesenchymal condensation

KW - Prx

UR - http://www.scopus.com/inward/record.url?scp=84955091855&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84955091855&partnerID=8YFLogxK

U2 - 10.1073/pnas.1515397113

DO - 10.1073/pnas.1515397113

M3 - Article

C2 - 26721400

AN - SCOPUS:84955091855

VL - 113

SP - E338-E347

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

SN - 0027-8424

IS - 3

ER -