1. Biochemistry and Chemical Biology
  2. Immunology and Inflammation

Mechanism of bisphosphonate-related osteonecrosis of the jaw (BRONJ) revealed by targeted removal of legacy bisphosphonate from jawbone using equilibrium competing inert hydroxymethylene diphosphonate

  1. Hiroko Okawa
  2. Takeru Kondo
  3. Akishige Hokugo  Is a corresponding author
  4. Philip Cherian
  5. Jesus J Campagna
  6. Nicholas A Lentini
  7. Eric C Sung
  8. Samantha Chiang
  9. Yi-Ling Lin
  10. Frank H Ebetino
  11. Varghese John
  12. Shuting Sun  Is a corresponding author
  13. Charles E McKenna  Is a corresponding author
  14. Ichiro Nishimura  Is a corresponding author
  1. University of California, Los Angeles, United States
  2. BioVinc, United States
  3. University of Southern California, United States
https://doi.org/10.7554/eLife.76207
Share this article
Cite this article
  1. Hiroko Okawa
  2. Takeru Kondo
  3. Akishige Hokugo
  4. Philip Cherian
  5. Jesus J Campagna
  6. Nicholas A Lentini
  7. Eric C Sung
  8. Samantha Chiang
  9. Yi-Ling Lin
  10. Frank H Ebetino
  11. Varghese John
  12. Shuting Sun
  13. Charles E McKenna
  14. Ichiro Nishimura
(2022)
Mechanism of bisphosphonate-related osteonecrosis of the jaw (BRONJ) revealed by targeted removal of legacy bisphosphonate from jawbone using equilibrium competing inert hydroxymethylene diphosphonate
eLife 11:e76207.
https://doi.org/10.7554/eLife.76207
  2. Download BibTeX
  3. Download .RIS

Abstract

Bisphosphonate-related osteonecrosis of the jaw (BRONJ) presents as a morbid jawbone lesion in patients exposed to a nitrogen-containing bisphosphonate (N-BP). Although it is rare, BRONJ has caused apprehension among patients and healthcare providers and decreased acceptance of this anti-resorptive drug class to treat osteoporosis and metastatic osteolysis. We report here a novel method to elucidate the pathological mechanism of BRONJ by the selective removal of legacy N-BP from the jawbone using an intra-oral application of hydroxymethylene diphosphonate (HMDP) formulated in liposome-based deformable nanoscale vesicles (DNV). After maxillary tooth extraction, zoledronate-treated mice developed delayed gingival wound closure, delayed tooth extraction socket healing and increased jawbone osteonecrosis consistent with human BRONJ lesion. Single cell RNA sequencing of mouse gingival cells revealed oral barrier immune dysregulation and unresolved pro-inflammatory reaction. HMDP-DNV topical applications to nascent mouse BRONJ lesions resulted in accelerated gingival wound closure and bone socket healing as well as attenuation of osteonecrosis development. The gingival single cell RNA sequencing demonstrated resolution of chronic inflammation by increased anti-inflammatory signature gene expression of lymphocytes and myeloid-derived suppressor cells. This study suggests that BRONJ pathology is related to N-BP levels in jawbones and demonstrates the potential of HMDP-DNV as an effective BRONJ therapy.

Data availability

All data generated or analyzed during this study are included in the manuscript and supporting file. Single cell RNA sequencing data have been deposited in GEO under accession code GSE193110.

The following data sets were generated

Article and author information

Author details

  1. Hiroko Okawa

    Weintraub Center for Reconstructive Biotechnology, University of California, Los Angeles, Los Angeles, United States
    Competing interests
    No competing interests declared.

  2. Takeru Kondo

    Weintraub Center for Reconstructive Biotechnology, University of California, Los Angeles, Los Angeles, United States
    Competing interests
    No competing interests declared.

  3. Akishige Hokugo

    Department of Surgery, University of California, Los Angeles, Los Angeles, United States
    For correspondence
    ahokugo@mednet.ucla.edu
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-7097-3364

  4. Philip Cherian

    BioVinc, Pasadena, United States
    Competing interests
    Philip Cherian, is an employee in BioVinc LLC..

  5. Jesus J Campagna

    Department of Neurology, University of California, Los Angeles, Los Angeles, United States
    Competing interests
    No competing interests declared.

  6. Nicholas A Lentini

    Department of Chemistry, University of Southern California, Los Angeles, United States
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-1701-0753

  7. Eric C Sung

    Weintraub Center for Reconstructive Biotechnology, University of California, Los Angeles, Los Angeles, United States
    Competing interests
    No competing interests declared.

  8. Samantha Chiang

    Section of Oral Biology, University of California, Los Angeles, Los Angeles, United States
    Competing interests
    No competing interests declared.

  9. Yi-Ling Lin

    Section of Oral and Maxillofacial Pathology, University of California, Los Angeles, Los Angeles, United States
    Competing interests
    No competing interests declared.

  10. Frank H Ebetino

    BioVinc, Pasadena, United States
    Competing interests
    Frank H Ebetino, holds equity in BioVinc LLC and is an employee and has executive management positions in BioVinc LLC..

  11. Varghese John

    Department of Neurology, University of California, Los Angeles, Los Angeles, United States
    Competing interests
    No competing interests declared.

  12. Shuting Sun

    BioVinc, Pasadena, United States
    For correspondence
    shuting.sun@biovinc.com
    Competing interests
    Shuting Sun, holds equity in BioVinc LLC. SS is an employee and has executive management position in BioVinc LLC..

  13. Charles E McKenna

    Department of Chemistry, University of Southern California, Los Angeles, United States
    For correspondence
    mckenna@usc.edu
    Competing interests
    Charles E McKenna, is a board member and holds equity in Biovinc LLC..
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-3540-6663

  14. Ichiro Nishimura

    Weintraub Center for Reconstructive Biotechnology, University of California, Los Angeles, Los Angeles, United States
    For correspondence
    inishimura@dentistry.ucla.edu
    Competing interests
    Ichiro Nishimura, was a consultant of BioVinc LLC..
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-3749-9445

Funding

National Institute of Dental and Craniofacial Research (R01DE022552)

  • Ichiro Nishimura

National Institute of Dental and Craniofacial Research (R44DE025524)

  • Frank H Ebetino
  • Ichiro Nishimura

Tohoku University (Leading young researcher overseas visit program fellowship)

  • Hiroko Okawa

Japan Society for the Promotion of Science (19J117670)

  • Takeru Kondo

National Center for Research Resources (C06RR014529)

  • Ichiro Nishimura

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Ethics

Animal experimentation: All animal experiments were performed at UCLA. All the protocols for animal experiments were approved by the UCLA Animal Research Committee (ARC# 1997-136) and followed the Public Health Service Policy for the Humane Care and Use of Laboratory Animals and the UCLA Animal Care and Use Training Manual guidelines. The C57Bl/6J mice (Jackson Laboratory) were used in this study. Animals consumed gel or regular food for rodents and water ad libitum and were maintained in regular housing conditions with a 12-hour-light/dark cycles at the Division of Laboratory Animal Medicine at UCLA.

Human subjects: This study was not conducted on human subjects. However, the manuscript contains clinical demonstration of human BRONJ obtained from patients of UCLA School of Dentistry clinic with the general consent for educational use. The information was not part of investigator-initiated research.

Copyright

© 2022, Okawa et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

Metrics

  • 2,241
    views
  • 400
    downloads
  • 17
    citations

Views, downloads and citations are aggregated across all versions of this paper published by eLife.

Download links

A two-part list of links to download the article, or parts of the article, in various formats.

Downloads (link to download the article as PDF)

Open citations (links to open the citations from this article in various online reference manager services)

Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)

  1. Hiroko Okawa
  2. Takeru Kondo
  3. Akishige Hokugo
  4. Philip Cherian
  5. Jesus J Campagna
  6. Nicholas A Lentini
  7. Eric C Sung
  8. Samantha Chiang
  9. Yi-Ling Lin
  10. Frank H Ebetino
  11. Varghese John
  12. Shuting Sun
  13. Charles E McKenna
  14. Ichiro Nishimura
(2022)
Mechanism of bisphosphonate-related osteonecrosis of the jaw (BRONJ) revealed by targeted removal of legacy bisphosphonate from jawbone using equilibrium competing inert hydroxymethylene diphosphonate
eLife 11:e76207.
https://doi.org/10.7554/eLife.76207

Share this article

https://doi.org/10.7554/eLife.76207

Categories and tags

Research organism

Further reading

    1. Biochemistry and Chemical Biology

    Disordered proteins interact with the chemical environment to tune their protective function during drying

    Shraddha KC, Kenny H Nguyen ... Thomas C Boothby
    Research Article

    The conformational ensemble and function of intrinsically disordered proteins (IDPs) are sensitive to their solution environment. The inherent malleability of disordered proteins, combined with the exposure of their residues, accounts for this sensitivity. One context in which IDPs play important roles that are concomitant with massive changes to the intracellular environment is during desiccation (extreme drying). The ability of organisms to survive desiccation has long been linked to the accumulation of high levels of cosolutes such as trehalose or sucrose as well as the enrichment of IDPs, such as late embryogenesis abundant (LEA) proteins or cytoplasmic abundant heat-soluble (CAHS) proteins. Despite knowing that IDPs play important roles and are co-enriched alongside endogenous, species-specific cosolutes during desiccation, little is known mechanistically about how IDP-cosolute interactions influence desiccation tolerance. Here, we test the notion that the protective function of desiccation-related IDPs is enhanced through conformational changes induced by endogenous cosolutes. We find that desiccation-related IDPs derived from four different organisms spanning two LEA protein families and the CAHS protein family synergize best with endogenous cosolutes during drying to promote desiccation protection. Yet the structural parameters of protective IDPs do not correlate with synergy for either CAHS or LEA proteins. We further demonstrate that for CAHS, but not LEA proteins, synergy is related to self-assembly and the formation of a gel. Our results suggest that functional synergy between IDPs and endogenous cosolutes is a convergent desiccation protection strategy seen among different IDP families and organisms, yet the mechanisms underlying this synergy differ between IDP families.

    1. Biochemistry and Chemical Biology
    2. Structural Biology and Molecular Biophysics

    Isobaric crosslinking mass spectrometry technology for studying conformational and structural changes in proteins and complexes

    Jie Luo, Jeff Ranish
    Tools and Resources

    Dynamic conformational and structural changes in proteins and protein complexes play a central and ubiquitous role in the regulation of protein function, yet it is very challenging to study these changes, especially for large protein complexes, under physiological conditions. Here, we introduce a novel isobaric crosslinker, Qlinker, for studying conformational and structural changes in proteins and protein complexes using quantitative crosslinking mass spectrometry. Qlinkers are small and simple, amine-reactive molecules with an optimal extended distance of ~10 Å, which use MS2 reporter ions for relative quantification of Qlinker-modified peptides derived from different samples. We synthesized the 2-plex Q2linker and showed that the Q2linker can provide quantitative crosslinking data that pinpoints key conformational and structural changes in biosensors, binary and ternary complexes composed of the general transcription factors TBP, TFIIA, and TFIIB, and RNA polymerase II complexes.

    1. Biochemistry and Chemical Biology
    2. Stem Cells and Regenerative Medicine

    Proteomic and functional comparison between human induced and embryonic stem cells

    Alejandro J Brenes, Eva Griesser ... Angus I Lamond
    Research Article

    Human induced pluripotent stem cells (hiPSCs) have great potential to be used as alternatives to embryonic stem cells (hESCs) in regenerative medicine and disease modelling. In this study, we characterise the proteomes of multiple hiPSC and hESC lines derived from independent donors and find that while they express a near-identical set of proteins, they show consistent quantitative differences in the abundance of a subset of proteins. hiPSCs have increased total protein content, while maintaining a comparable cell cycle profile to hESCs, with increased abundance of cytoplasmic and mitochondrial proteins required to sustain high growth rates, including nutrient transporters and metabolic proteins. Prominent changes detected in proteins involved in mitochondrial metabolism correlated with enhanced mitochondrial potential, shown using high-resolution respirometry. hiPSCs also produced higher levels of secreted proteins, including growth factors and proteins involved in the inhibition of the immune system. The data indicate that reprogramming of fibroblasts to hiPSCs produces important differences in cytoplasmic and mitochondrial proteins compared to hESCs, with consequences affecting growth and metabolism. This study improves our understanding of the molecular differences between hiPSCs and hESCs, with implications for potential risks and benefits for their use in future disease modelling and therapeutic applications.