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

    64 Citations (Scopus)

    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

    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.
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    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.",
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    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

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